NL2037452B1

NL2037452B1 - Agents for treating Celiac Disease

Info

Publication number: NL2037452B1
Application number: NL2037452A
Authority: NL
Inventors: Koning Frits; Heidt Sebastiaan; H M Heemskerk Mirjam; Gille Ilse
Original assignee: Academisch Ziekenhuis Leiden
Priority date: 2024-04-12
Filing date: 2024-04-12
Publication date: 2025-11-03
Also published as: WO2025216635A1

Abstract

The invention relates to the field of novel chimeric HLA class II molecules, fusion polypeptides, nucleic acids, vectors, modified cells and compositions directed against celiac disease. Associated methods for use in treating or preventing celiac disease are also provided herein.

Description

Agents for treating Celiac Disease

The invention relates to the field of novel chimeric HLA class Il molecules, fusion polypeptides, nucleic acids, vectors, modified cells and compositions directed against celiac disease. Associated methods for use in treating or preventing celiac disease are also provided herein.

Background

Celiac disease (CD), also known as coeliac disease or celiac spruce, is a small intestinal disease caused by a pro-inflammatory T cell response to the food antigen gluten.

Celiac disease affects between 0.5 to 2% of the population in Europe, the America's, the

Middle East, Northern Africa, India, China and Australia. At present, the only available treatment for celiac disease is a strict lifelong gluten-free diet which is complicated by the widespread use of gluten in the food industry, wheat being one of the most commonly consumed cereals worldwide. Gluten and the gluten-like proteins hordein and secalin are an integral component of wheat, barley and rye respectively. As such, they are an essential component of commonly consumed foods including but not restricted to bread, pasta, and pizza. In addition, gluten is often added to food products not readily associated with cereals like soy sauce, instant meals and soups.

Celiac disease occurs in subjects possessing either HLA-DQ2 and/or HLA-DQ8. HLA-

DQ molecules bind to peptides derived from endogenous or exogenous sources and display

HLA-DQ:peptide complexes on the surface of antigen presenting cells. If the peptides are derived from a pathogen, HLA-DQ:peptide complexes are recognized by T cell receptors (TCRs) expressed by CD4+ T cells. CD4+ T cells specifically interact with a HLA-DQ: peptide complex, leading to T cell activation and the mounting of an immune response to eradicate the pathogen.

HLA-DQ2 and HLA-DQ8 are uniquely suited to bind proline-rich and transglutaminase (TG2) modified gluten-derived peptides. HLA-DQ2 and HLA-DQ8 have a strong preference for negatively charged amino acids at particular positions in the bound peptide that serve to anchor the bound peptide tightly into the peptide-binding groove. While gluten itself does not contain such negatively charged amino acids, negatively charged amino acids are incorporated into gluten peptides in the small intestine to form gluten-derived peptides. When gluten peptides enter the small intestine, the enzyme tissue TG2 converts glutamine residues in the gluten peptides into glutamic acid, thus producing proline-rich and TG2 modified gluten- derived peptides.

Due to their strong association, HLA-DQ2 and HLA-DQ8 bind to said proline-rich and

TG2 modified gluten-derived peptides, forming HLA-DQ:gluten-derived peptide complexes.

CD4+ T cells expressing T cell receptors bind specifically to these HLA-DQ:gluten-derived peptide complexes, resulting in T cell activation and the pro-inflammatory phenotype found in subjects with celiac disease.

Gluten is a complex mixture of proteins consisting of gliadins and glutenin, where the gliadins can be subdivided in a-, y-, and w-gliadins and the glutenins in the (high molecular weight) HMW-glutenins and low molecular weight (LMW)-glutenins. Although immunogenic peptides can be found in all of these proteins classes, the T cell response in HLA-DQ2 positive celiac disease subjects is directed to peptide sequences in the N-terminal part of a-gliadin and homologous sequences thereof in w-gliadin. The w-gliadin sequences are identical to sequences found in the hordeins of barley and the secalins of rye. Similarly, in HLA-DQ8 positive celiac disease subjects, the T cell response is directed to peptide sequences in the

C-terminal part of a-gliadin.

Therefore, the T cell response in subjects with celiac disease is focussed on a-gliadin, w-gliadin, hordein and secalin derived peptides. It has been shown that such T cells can persist for decades, providing a likely explanation for the lifelong requirement of a gluten free diet.

Accordingly, there is a need for preventing and/or eliminating this T cell response, and thus provide an improved means for treating celiac disease.

Summary of the Invention

The invention provides a chimeric HLA class II molecule comprising: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8182 chain and (ily a transmembrane domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

B1B2 chains are encoded by the genes HLA-DQA 1*03 and HLA-DQB1*03, respectively.

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising:

(i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3, SEQ ID NO: 4 and SEQ ID NO: 5 (preferably SEQ ID NO: 5) and an HLA-DQ B1B2 chain and (ii) a transmembrane domain (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and

B1B2 chains are encoded by the genes HLA-DQA 1*03 and HLA-DQB1*03, respectively. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3, SEQ ID NO: 4 and SEQ ID NO: 5 (preferably SEQ ID NO: 5) and an HLA-DQ B1B2 chain (ii) a transmembrane domain and (iiiy an immune receptor intracellular signaling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

B1B2 chains are encoded by the genes HLA-DQA1*03 and HLA-DQB1*03, respectively. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 46) and an HLA-DQ 8182 chain and

(ii) a transmembrane domain {b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 46) and an HLA-DQ B12 chain (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA 1*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

B1B2 chains are encoded by the genes HLA-DQA 1*03 and HLA-DQB 103, respectively. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3. SEQ ID NO:4, SEQ ID NO: 5, SEQ ID

NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 5 and SEQ ID NO: 46) and an HLA-DQ 8182 chain and (ii) a transmembrane domain (b) a second polypeptide comprising:

(i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; 5 wherein the HLA-DQ a1a2 and B182 chains are encoded by the genes HLA-DQA1*05 and

NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 5 and SEQ ID NO: 46) and an HLA-DQ B1B2 chain (iù a transmembrane domain and (iii) an immune receptor intracellular signaling domain; and {b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

B1B2 chains are encoded by the genes HLA-DQA 103 and HLA-DQB1*03, respectively. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3, SEQ ID NO: 4 and SEQ ID NO: 5 (preferably SEQ ID NO: 5) and an HLA-DQ B1B2 chain and (ii) a transmembrane domain (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively. Optionally, the HLA-DQB1*02 is HLA-DQB1*02:01 and the HLA-

DQAT*05 is HLA-DQA1*05:01. See for example SEQ ID NO: 13 and SEQ ID NO: 7. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3, SEQ ID NO: 4 and SEQ ID NO: 5 (preferably SEQ ID NO: 5) and an HLA-DQ 8182 chain (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

DQA1*05 is HLA-DQA1*05:01. See for example SEQ ID NO: 13 and SEQ ID NO: 7. The transmembrane domain may be CD28. The molecule may further comprise a co-stimulatory domain (e.g. CD28) and/or a primary signalling domain (e.g. CD3 zeta).

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 46) and an HLA-DQ 8182 chain and (ii) a transmembrane domain (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively. Optionally, the HLA-DQB1*02is HLA-DQB1*02:01 and the HLA-

Suitably, the chimeric HLA class II molecule may comprise: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 46) and an HLA-DQ B1B2 chain (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQBT*02, respectively. Optionally, the HLA-DQB1*02 is HLA-DQB1*02:01 and the HLA-

NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 5 and SEQ ID NO: 48) and an HLA-DQ B1B2 chain and (ii) a transmembrane domain {b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

(i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes selected from SEQ ID NO:3. SEQ ID NO:4, SEQ ID NO: 5, SEQ ID

NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46 (preferably SEQ ID NO: 5 and SEQ ID NO: 48) and an HLA-DQ B12 chain (ii) a transmembrane domain and (ii an immune receptor intracellular signaling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain and (ii) a transmembrane domain and (iii) an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

Fusion polypeptides, nucleic acids, vectors, modified cells and compositions directed against celiac disease corresponding to the above are also provided herein.

The invention provides a fusion polypeptide, comprising, in an N-terminal to C-terminal orientation: (a) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8182 chain; {b) a transmembrane domain; ({c) a peptide cleavage signal; (d) a second extracellular domain comprising an HLA-DQ a1a2 chain; (e) a transmembrane domain; wherein the polypeptide further comprises an immune receptor intracellular signalling domain located at the C-terminus of (b) and/or (e), and wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA 102 and HLA-DQB 1702, respectively; or wherein the HLA-DQ a1a2 and

B1B2 chains are encoded by the genes HLA-DQA 1*03 and HLA-DQB 1703, respectively.

Suitably, the fusion polypeptide may further comprise an HLA-DQ B1B2 chain signal sequence located N-terminal to the first extracellular domain; and an HLA-DQ a1a2 chain signal sequence located N-terminal to the second extracellular domain.

Suitably, the one or more gluten-derived epitopes may be from a protein selected from the group consisting of: gliadin, glutenin, hordein, secalin and avenin.

Suitably, the gliadin may be alpha gliadin or omega gliadin.

Suitably, the gliadin may be alpha gliadin.

Suitably, the one or more gluten-derived epitopes may comprise the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 69.

Suitably, the one or more gluten-derived epitopes may comprise the amino acid sequence of SEQ ID NO: 61.

Suitably, the gluten-derived peptide may comprise two or more gluten-derived epitopes.

Suitably, the gluten-derived peptide may comprise the amino acid sequences of SEQ

ID NO: 3 and SEQ ID NO: 4; or SEQ ID NO: 69 and SEQ ID NO:4.

Suitably, the gluten-derived peptide may comprise the amino acid sequence of SEQ

ID NO: 5 or SEQ ID NO: 70.

Suitably, the gliadin may be omega gliadin.

Suitably, the one or more gluten-derived epitopes may comprise the amino acid sequence of SEQ ID NO: 44.

Suitably, the one or more gluten-derived epitopes may comprise the amino acid sequence of SEQ ID NO: 45.

ID NO: 44 and SEQ ID NO: 45.

ID NO: 46.

Suitably, the gluten-derived peptide may comprise four or more gluten-derived epitopes.

Suitably, the gluten-derived peptides may be selected from the group consisting of

SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 69, SEQ ID NO: 44 and SEQ ID NO: 45.

Suitably, the gluten-derived peptide may comprise: (a) the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 46, optionally wherein the gluten-derived peptide comprises a linker between the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 48, or (b) the amino acid sequence of SEQ ID NO: 70 and SEQ ID NO: 46, optionally wherein the gluten-derived peptide comprises a linker between the amino acid sequence of SEQ ID NO: 70 and SEQ ID NO: 46.

Suitably, the first extracellular domain may further comprise a linker sequence between the gluten-derived peptide and the HLA-DQ B1B2 chain.

Suitably, the gluten-derived peptide may further comprise flanking amino acids at the

N- and/or C- terminal side of the gluten-derived epitope, optionally wherein the flanking amino acids are natural flanking amino acids.

Suitably, the transmembrane domain may be selected from the group consisting of: alpha or beta chain of CD28, CD4, CD5, CD8, CD9, CD16, CD22, CD27, CD33, CD37, CD45,

CDe4, CD80, CD86, CD134, CD137, CD152, CD154, PD1, HLA-DR, HLA-DQ or HLA-DP.

Suitably, the transmembrane domain may be a CD28 transmembrane domain.

Suitably, the transmembrane domain may be a HLA-DQ transmembrane domain.

Suitably, the immune receptor intracellular signalling domain may comprise one or more co-stimulatory signalling domains.

Suitably, the one or more co-stimulatory signalling domain may be selected from the group consisting of: TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLRS, TLR10,

CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (0X40), CD137 (4-1BB), CD154 (CD40L), CD278 (ICOS), DAP10, LAT, NKD2C, SLP76, TRIM, and ZAP70 co-stimulatory signalling domain.

Suitably, the one or more co-stimulatory signalling domain may be a CD28 co- stimulatory signalling domain.

Suitably, the one or more co-stimulatory signalling domain may be a CD137 co- stimulatory signalling domain.

Suitably, the immune receptor intracellular signalling domain may comprise a primary signalling domain.

Suitably, the primary signalling domain may be selected from the group consisting of:

FcRy, FcRB, CD3y, CD3ò, CD3e, CD3¢, CD22, CD79b, and CD66d.

Suitably, the primary signalling domain may beCD3Z.

The invention provides a nucleic acid encoding a chimeric HLA class II molecule disclosed herein, or a fusion polypeptide disclosed herein, optionally wherein the nucleic acid is RNA and/or DNA.

The invention provides a vector comprising a nucleic acid disclosed herein.

The invention provides a cell comprising one or more nucleic acid, vector, chimeric

HLA class Il molecule, and/or fusion polypeptide disclosed herein.

Suitably, the cell may be a T cell, optionally wherein the T cell is selected from the group consisting of a CD8+ T cell, an NK T cell, CD3+ T cell and yd T cell, or a mixture of any one thereof.

Suitably, the cell may be a NK cell or an innate lymphoid cell (ILC).

The invention provides a cell comprising at least two of the following: a) a first nucleic acid, vector, chimeric HLA class Il molecule, or fusion polypeptide disclosed herein, wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-

DQAT*05 and HLA-DQB1*02, respectively;

b) a second nucleic acid, vector, chimeric HLA class Il molecule, or fusion polypeptide disclosed herein wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-

DQA1*02 and HLA-DQB1*02, respectively; and/or c) a third nucleic acid, vector, chimeric HLA class Il molecule, or fusion polypeptide disclosed herein, wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-

DQA1*03 and HLA-DQB1*03, respectively.

Suitably, the HLA-DQA1*02 and HLA-DQB1*02 may be HLA-DQA1*02:01 and HLA-

DQB71*02:02 respectively.

Suitably, the HLA-DQA1*05 and HLA-DQB1*02 may be HLA-DQA1*05:01 and HLA-

DQB1*02:01 respectively.

The invention provides a composition comprising a nucleic acid, vector, chimeric HLA class II molecule, fusion polypeptide and/or cell disclosed herein.

The invention provides a pharmaceutical composition comprising a nucleic acid, vector, chimeric HLA class Il molecule, fusion polypeptide and/or cell disclosed herein.

The invention provides a pharmaceutical composition disclosed herein, for use as a medicament.

The invention provides a pharmaceutical composition disclosed herein, for use in treating or preventing celiac disease in a HLA-DQ positive subject.

Suitably, the subject may be HLA-DQA1*02-HLA-DQB1*02, HLA-DQA1*05-HLA-

DQB1*02, and/or a HLA-DQA1*03-DQB1*03 positive.

Brief Description of the Figures

Embodiments of the invention are further described hereinafter with reference to the accompanying drawings, in which:

Figures 1A to B: Different chimeric HLA class Il molecule constructs (also referred to as CHAR constructs/CHAR molecules herein). (A) Graphical representation of a CHAR T cell expressing an HLA-DQ2 CHAR molecule, either linked with gliadin peptide or alternative CLIP peptide; (B) Schematic overview of the HLA-DQ2-gluten or HLA-DQ2-CLIP

CHAR constructs; the sequences of the specific peptides are linked to the extracellular part of an HLA-DQB1*02:01 molecule coupled to the CD28 costimulatory transmembrane and intracellular signaling domain, and CD3( intracellular signaling domain, coupled to P2A, and the extracellular part of an HLA-DQA1*05:01 molecule coupled to the CD28-CD34, followed by IRES allowing for co-expression of NGFR.

Figures 2A to D: Gliadin-specific T cell clones produce IFNy and IL-2 upon HLA-

DQ2 CHAR stimulation. (A) IL-2 release by S2 and S18 clones when incubated with K562 cells expressing different HLA-DQ2 CHAR constructs. In Figure A, for K562+DQ2-glia-a

CHAR, the left bar is S2 and the right bar is S16; for CD3/CD28 beads, the left bar is S2,

middle bar is S18, and right bar is SV30 (B) and at different effector: target ratios. In Figure B, for each ratio, the left bar is S2 and the right bar is S16 (C) IFNy release by S2 and S16 clones when incubated with K562 cells expressing different HLA-DQ2 CHAR constructs. In Figure C, for K562+DQ2-glia-a CHAR, the left bar is S2 and the right bar is S16; for CD3/CD28 beads, the left bar is S2, middle bar is S16, and right bar is SV30 {D} and at different effector: target ratios. In Figure D, for each ratio, the left bar is S2 and the right bar is S16.

Figures 3A to C: Specific lysis of gliadin-specific T cell clones after incubation with HLA-DQ2 CHAR T cells. (A) The percentage of specific lysis by HLA-DQ2-glia-a CHAR (circles), HLA-DQ2-CLIP CHAR (squares), or mock (triangles) T cells of glia-a1-specific S2 clone at different effector: target (E: T) ratios shown in two different donors; LRG (A1) and

UTT (A2). (B) The percentage of specific lysis by HLA-DQ2-glia-a CHAR (circles), HLA-DQ2-

CLIP CHAR (squares), or mock (triangles) T cells of glia- a2-specific S16 clone at different effector: target (E: T) ratios shown in two different donors; LRG (B1) and UTT (B2). (C) The percentage of specific lysis by HLA-DQ2-glia-a CHAR (circles), HLA-DQ2-CLIP CHAR (squares), or mock (triangles) T cells of glia-y2-specific SV30 clone at different effector: target (E: T) ratios shown in two different donors; LRG (C1) and UTT (C2).

The patent, scientific and technical literature referred to herein establish knowledge that was available to those skilled in the art at the time of filing. The entire disclosures of the issued patents, published and pending patent applications, and other publications that are cited herein are hereby incorporated by reference to the same extent as if each was specifically and individually indicated to be incorporated by reference. In the case of any inconsistencies, the present disclosure will prevail.

Various aspects of the invention are described in further detail below.

Definitions “HLA” and “human leukocyte antigen” are herein defined as a genetic fingerprint on human nucleated cells and platelets, composed of proteins that play a critical role in activating the body's immune system to respond to foreign organisms. In humans and other animals, the

HLA is also referred to as the “major histocompatibility complex” (MHC). HLA class Il is encoded by three different isotypes, HLA-DR, -DQ, and -DP, and presents antigens (typically peptides) from outside a cell to T-lymphocytes. These particular antigens stimulate multiplication of T-helper cells (also called CD4-positive T cells), which in turn stimulate antibody-producing B-cells to produce antibodies to specific antigens and stimulate cytotoxic

T cells to lyse infected cells. Natural HLA class II molecules are heterodimers with two homologous chains, the a and B chains. The a chain is made up of an a1 (antigen-binding e.g. gluten-derived peptide binding) domain, an a2 (conserved) domain, and a transmembrane domain. The B chain is made up of a 1 {antigen-binding e.g. gluten-derived peptide binding)

domain, a B2 (conserved) domain, and a transmembrane domain. The a and B chains of HLA class II (HLA-II) molecules or proteins are capable of specifically binding (and therefore presenting) a peptide antigen derived from extracellular proteins, including those of extracellular pathogens, on the cell surface. “Chimeric HLA class Il molecule” (referred to herein as “the molecule”, or “chimeric

HLA antigen receptor”, or “CHAR” herein) refers to an HLA class Il molecule which comprises an HLA-DQ a1a2 chain and a HLA-DQ B1B2 chain, wherein the HLA-DQ B1B2 chain is fused to a gluten-derived peptide. Suitably, the gluten-derived peptide may be presented in the HLA groove of the molecule. Suitable gluten-derived peptides are described elsewhere herein.

Suitably, the HLA-DQ a1a2 chain and/or the HLA-DQ B1B2 chain of the molecule may be fused to a transmembrane domain and one or more intracellular signalling domains (optionally containing co-stimulatory domain(s) (see, e.g., Sadelain et al, Cancer Discov., 3(4):388 (2013); see also Harris and Kranz, Trends Pharmacol. Sci., 37(3):220 (2016), and Stone et al,

Cancer Immunol. Immunother., 63(11): 1163 (2014)). In the context of the present disclosure, a chimeric HLA class Il molecule that presents the peptide fused to the HLA-DQ B1B2 chain may be referred to as a chimeric HLA class ll:peptide complex (where the fused peptide is presented in the HLA groove, thus forming an HLA class Il:peptide complex). Accordingly, a chimeric HLA class Il molecule may also be referred to herein as a chimeric HLA class

Il:peptide complex (wherein the gluten-derived peptide is presented within the HLA groove generated by the HLA-DQ a1a2 chain and the HLA-DQ 8182 chain).

The term “extracellular domain” refers to polypeptide domain that, when expressed by a cell, is located at the external surface of a cell. The extracellular domains of the invention comprise the extracellular components of HLA class II molecules, for example, the a1a2 chain and/or B1B2 chain of HLA-DQ2 or -DQ8. The extracellular domain may be derived either from a natural, synthetic, semi-synthetic, or recombinant source.

As used herein "specifically binds" or "specific for" refers to an association or union of a binding protein (e.g., a chimeric HLA class Il molecule and/or TCR receptor) or a binding domain (or fusion protein thereof) to a target molecule with an affinity or Kas (i.e., an equilibrium association constant of a particular binding interaction with units of 1/M) equal to or greater than 105M7 (which equals the ratio of the on-rate [kon] to the off-rate [kor] for this association reaction), while not significantly associating or uniting with any other molecules or components in a sample. Binding proteins or binding domains (or fusion proteins thereof) may be classified as "high affinity" binding proteins or binding domains {or fusion proteins thereof) or as "low affinity" binding proteins or binding domains (or fusion proteins thereof). "High affinity” binding proteins or binding domains refer to those binding proteins or binding domains having a Ka of at least 107 Mt, at least 108 Mt, at least 10° Mt, at least 1019 M1, at least 10°! MM, at least 1012

Mt, or at least 103 M’. Low affinity” binding proteins or binding domains refer to those binding proteins or binding domains having a Ka of up to 107 M* up to 10° Mt, up to 105 M™.

Alternatively, affinity can be defined as an equilibrium dissociation constant (Ka) of a particular binding interaction with units of M (e.g., 10°Mto 10" M).

In certain embodiments, a receptor or binding domain may have "enhanced affinity," which refers to selected or engineered receptors or binding domains with stronger binding to a target antigen than a wild type (or parent) binding domain. For example, enhanced affinity may be due to a Ks (equilibrium association constant) for the target antigen that is higher than the wild type binding domain, due to a K4 (dissociation constant) for the target antigen that is less than that of the wild type binding domain, due to an off-rate (kos) for the target antigen that is less than that of the wild type binding domain, or a combination thereof.

The term “gluten-derived peptide” refers to peptides derived from, or encompassed within, one or more gluten proteins. A gluten protein may be selected from the group consisting of gliadin, glutenin, hordein, secalin and avenin (see, for example, Sollid LM, Tye-Din JA, Qiao

SW, Anderson RP, Gianfrani C, Koning F. Update 2020: nomenclature and listing of celiac disease-relevant gluten epitopes recognized by CD4+ T cells. Immunogenetics. 2019 Nov 18).

Tissue transglutaminase (TG2) can modify proteins by transamidation or deamidation of specific glutamine residues. As would be appreciated by a person of skill in the art, in vivo,

TG2 causes selective deamidation of gluten, which in turn, causes the generation of a series of proline-rich and TG2 modified gluten-derived peptides that bind to HLA-DQ2 or -DQ8 molecules with high affinity. In the context of the invention, “gluten-derived peptides” may also be referred to as “gluten peptide or deaminated components thereof’, “deaminated gluten peptide”, “TG2 modified gluten peptide” or “proline-rich gluten peptide”. Similarly, the term “gluten-derived epitope” may also be referred to as “gluten epitope or deaminated components thereof”, “deaminated gluten epitope”, “TG2 modified gluten epitope” or “proline-rich gluten epitope”.

As used herein, “gliadin” refers to the aqueous alcohol-soluble fraction of gluten, particularly, but not exclusively, gluten derived from wheat, for example Triticum aestivum. A gliadin protein may be selected from the group consisting of alpha (a), beta (8), gamma (y) and omega (w) gliadin.

As used herein, “glutenin” refers to the aqueous alcohol-insoluble fraction of gluten, particularly but not exclusively, gluten derived from wheat, for example Triticum aestivum.

Glutenin may be selected from the group consisting of low and high molecule weight (LMW and HMW) glutenin.

As used herein, “hordein” or “barley hordein” refers to gluten derived from barley, Hordein vulgare. Hordein may be selected from the group consisting of B hordein, C hordein and D hordein.

As used herein, “secalin” or “rye secalin” refers to gluten derived from rye, Secale cerale. Secalin may be selected from the group consisting of B secalin, y secalin and w secalin.

As used herein “avenin” refers to gluten derived from oats, Avena sativa.

The term “peptide” as used herein refers to a polymer of amino acids. The peptide may be relatively short (i.e. no more than 20 amino acids; e.g. no more than 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, or 8 amino acids).

As used herein, “epitope” refers to that portion of an antigen or a peptide that is recognized by the immune system, for example, a T cell receptor or HLA class | or class Il, an antibody, a B cell receptor, which portion is sufficient for high affinity binding.

As used herein, “transmembrane domain” (TM domain or TMD; refers to a domain that anchors a polypeptide to the plasma membrane of a cell. The TM domain may be derived either from a natural, synthetic, semi-synthetic, or recombinant source.

As used herein “immune receptor intracellular signaling domain” is interchangeable with “intracellular signaling domain” or “endodomain”, and refers to the portion of a protein which transduces the effector function signal and that directs the cell to perform a specialized function. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire domain. To the extent that a truncated portion of an intracellular signaling domain is used, such truncated portion may be used in place of the entire domain as long as it transduces the effector function signal. The term intracellular signaling domain is meant to include any truncated portion of the intracellular signaling domain sufficient to transducing effector function signal.

As used herein “HLA-DQA1*02-HLA-DQB1*02” is interchangeable with HLA-DQ2 and refers to an HLA molecule encoded by the genes HLA-DQA1*02 and HLA-DQB1*02.

More specifically, in HLA-DQ2, the a1a2 chain is encoded by the gene HLA-DQA 1*02 and the

B1B2 chain is encoded by HLA-DQB1*02.

Suitably, the HLA-DQ2 molecule may be an HLA-DQ2.5 or HLA-DQ2.2 molecule.

A HLA-DQ2.5 molecule comprises an a1a2 chain encoded by the gene HLA-

DQA1*05:01 and a B1B2 chain encoded by the gene HLA-DQB1*02:01. Herein, the HLA-

DQ2.5 molecule may also be referred to as “HLA-DQA1*05:01-HLA-DQB1*02:01”.

Accordingly, a subject that is HLA-DQ2.5 positive is a subject that comprises HLA-DQ2.5, and may be referred to herein as an “HLA-DQA1*05:01-HLA-DQB1*02:01" positive subject.

An HLA-DQ2.2 molecule comprises an a1a2 chain encoded by the gene HLA-

DQA1*02:01 and a B1B2 chain encoded by the gene HLA-DQB1*02:02. Herein, the HLA-

DQ2.2 molecule may also be referred to as “HLA-DQA1*02:01-HLA-DQB1*02:02”.

Accordingly, a subject that is HLA-DQ2.2 positive is a subject that comprises HLA-DQ2.2, and may be referred to herein as an “HLA-DQA 1*02:01-HLA-DQB1*02:02” positive subject.

An HLA-DQ8 molecule comprises an ala2 chain encoded by the gene HLA-

DQA1*03 and a 8182 chain encoded by the gene HLA-DQB1*03. Suitably the HLA-DQA1*03 gene encoding the a1a2 chain may be HLA-DQA1*03:01 or HLA-DQA1*03:02. Suitably the

HLA-DQB1*03 gene encoding the B1B2 chain may be HLA-DQB1*03:02. Herein, the HLA-

DQ8 molecule may also be referred to as “HLA-DQA1*03:01-HLA-DQB1*03” or “HLA-

DQA1*03:02-HLA-DQB1*03”. Accordingly, a subject that is HLA-DQ8 positive is a subject that comprises HLA-DG8, and may be referred to herein as an “HLA-DQA1*03-HLA-DQB1*03 positive” subject.

As used herein, “fusion polypeptide” refers to one or more polypeptide domains or segments. Fusion polypeptides are typically linked N-terminus to C-terminus, although they can also be linked C-terminus to C-terminus, N-terminus to N- terminus, or C-terminus to N- terminus. In particular embodiments, the polypeptides of the fusion protein can be in any order.

Fusion polypeptides or fusion proteins can also include conservatively modified variants, polymorphic variants, alleles, mutants, subsequences, and interspecies homologs, so long as the desired activity of the fusion polypeptide is preserved. Fusion polypeptides may be produced by chemical synthetic methods or by chemical linkage between the two moieties or may generally be prepared using other standard techniques.

As used herein, “peptide cleavage signal” is interchangeable with “polypeptide cleavage site” and “protease cleavage sites”, and refers to a polypeptide cleavage signal between each of the polypeptide domains described herein. In addition, a polypeptide cleavage site can be put into any linker peptide sequence. Exemplary polypeptide cleavage signals include polypeptide cleavage recognition sites such as protease cleavage sites, nuclease cleavage sites (e.g., rare restriction enzyme recognition sites, self-cleaving ribozyme recognition sites), and self-cleaving viral oligopeptides (see deFelipe and Ryan, 2004. Traffic, 5(8); 616-26).

As used herein, “linker sequence” is interchangeable with “linker” and refers to a plurality of amino acid residues between the various polypeptide domains added for appropriate spacing and conformation of the molecule.

As used herein, “immunodominant” refers to a peptide stimulating immune responses to a greater extent than other peptides.

As used herein, “flanking amino acids” is interchangeable with “flanking sequences” and refers to amino acid residues adjacent to a specific amino acid sequence of interest. In the context of the present disclosure, the specific amino acid sequence of interest may be the gluten-derived peptide. Accordingly, the flanking amino acids may be those adjacent to the gluten-derived peptide. The flanking amino acids may be adjacent to the N- or C-terminal of the peptide. By being “adjacent” as used herein means abutting or in close proximity to the amino acid sequence of interest (for example no more than 1 or 2 amino acid residues away from the amino acid sequence of interest. More suitably, the flanking amino acid abuts the amino acid sequence of interest. Merely by way of example, the flanking amino acid may be 1, 2, 3, 4, or 5 amino acids in length. More suitably, the flanking amino acid may be 1 or 2 amino acids in length.

As used herein, the term, “co-stimulatory signaling domain,” or “co-stimulatory domain” refers to an intracellular signaling domain of a co-stimulatory molecule. Co- stimulatory molecules are cell surface molecules other than antigen receptors or Fc receptors that provide a second signal required for efficient activation and function of T lymphocytes upon binding to antigen.

As used herein “primary signaling domain” refers to an intracellular signaling domain that regulates the primary activation of the chimeric HLA class II molecule either in a stimulatory way, or in an inhibitory way. Primary signaling domains that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs or ITAMSs.

As used herein “nucleic acid sequence”, “polynucleotide”, “nucleic acid” and “nucleic acid molecule” are used interchangeably to refer to an oligonucleotide sequence or polynucleotide sequence. The nucleotide sequence may be of genomic, synthetic or recombinant origin, and may be double-stranded or single-stranded (representing the sense or antisense strand). The term "nucleotide sequence" includes genomic DNA, cDNA, synthetic

DNA, and RNA (e.g. mRNA) and analogs of the DNA or RNA generated, e.g., by the use of nucleotide analogs.

As used herein, “isolated nucleic acid sequence” or “isolated nucleic acid composition” refers to a nucleic acid sequence that is not in its natural environment when it is linked to its naturally associated sequence(s) that is/are also in its/their natural environment.

In other words, an isolated nucleic acid sequence/composition is not a native nucleotide sequence/composition, wherein "native nucleotide sequence/composition” means an entire nucleotide sequence that is in its native environment and when operatively linked to an entire promoter with which it is naturally associated, which promoter is also in its native environment.

Such a nucleic acid could be part of a vector and/or such nucleic acid or polypeptide could be part of a composition (e.g., a cell lysate), and still be isolated in that such vector or composition is not part of the natural environment for the nucleic acid or polypeptide. The term "gene" means the segment of DNA involved in producing a polypeptide chain; it includes regions preceding and following the coding region ("leader and trailer") as well as intervening sequences (introns) between individual coding segments (exons).

The nucleic acid sequences of the invention may be a non-naturally occurring nucleic acid sequence (e.g. it may be that the entire sequence does not occur in its entirety in nature).

For example, the nucleic acid sequence of the invention may be operably linked to a promoter,

wherein the promoter is not naturally associated with equivalent human nucleic acid sequences in nature (e.g. HLA sequences or fragments thereof); i.e. it is not the entire promoter that is naturally associated with the nucleic acid in its natural environment. In this context, such promoters may be considered exogenous promoters.

In some examples, a protein and a gene encoding said protein may be referred to using the same term (e.g. HLA-DQZ2). In examples where a protein and a gene encoding said protein are referred to using the same term (e.g. HLA-DQ2 etc), a person of skill in the art would readily be able to determine whether the protein or the gene was being referred to depending on the context in which the term was mentioned.

As used herein, the term “vector” refers to a nucleic acid sequence capable of transporting another nucleic acid sequence to which it has been operably linked. The vector can be capable of autonomous replication or it can integrate into a host DNA. The vector may include restriction enzyme sites for insertion of recombinant DNA and may include one or more selectable markers or suicide genes. The vector can be a nucleic acid sequence in the form of a plasmid, a bacteriophage or a cosmid.

As used herein, the term "cell" is interchangeable with “host cell” or “modified cell” and includes any cell into which the nucleic acid, vector, CHAR, fusion polypeptide described herein may be introduced. Once a nucleic acid, vector, CHAR, fusion polypeptide has been introduced into the cell, it may be referred to as a “modified cell” herein. Once the nucleic acid, vector, CHAR, fusion polypeptide is introduced into the host cell, the resultant modified cell should be capable of expressing the encoded binding protein (and e.g. correctly localising the encoded binding protein for its intended function e.g. transporting the encoded binding protein to the cell surface).

As used herein, a pharmaceutical composition may comprise a nucleic acid, vector,

CHAR or fusion polypeptide described herein along with a pharmaceutically acceptable excipient, adjuvant, diluent and/or carrier.

As used herein, "pharmaceutically acceptable" refers to a material that is not biologically or otherwise undesirable, i.e., the material may be administered to an individual along with the selected nucleic acid, vector, CHAR or fusion polypeptide without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. Pharmaceutically acceptable excipients are well known in the art. A suitable excipient is therefore easily identifiable by one of ordinary skill in the art. By way of example, suitable pharmaceutically acceptable excipients include water, saline, aqueous dextrose, glycerol, ethanol, and the like.

As used herein, the terms “treat”, “treating” and "treatment" are taken to include an intervention performed with the intention of preventing the development or altering the pathology of a condition, disorder or symptom (e.g. celiac disease or a celiac disease related condition). Accordingly, "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted condition, disorder or symptom. “Treatment” therefore encompasses a reduction, slowing or inhibition of the amount or concentration of target cells, for example as measured in a sample obtained from the subject, of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% when compared to the amount or concentration of target cells before treatment.

Methods of measuring the amount or concentration of target cells include, for example, qRT-

PCR, and quantification of disease specific biomarkers in a sample obtained from the subject.

As used herein the term “subject” refers to an individual, e.g., a human, having or at risk of having a specified condition, disorder or symptom. The subject may be a patient i.e. a subject in need of treatment in accordance with the invention. The subject may have received treatment for the condition, disorder or symptom. Alternatively, the subject has not been treated prior to treatment in accordance with the present invention.

As used herein, the term “HLA-DQ positive” refers to a subject who possess either

HLA-DQ2 or HLA-DG8. Such individuals mount an inappropriate HLA-DQ2- and/or DQ8- restricted CD4+ T cell-mediated immune response to gluten-derived peptides.

Detailed Description

The present invention provides a chimeric HLA class Il molecule comprising: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and a HLA-DQ 8182 chain and (ii) a transmembrane domain; and (b) a second polypeptide comprising: ( a second extracellular domain comprising a HLA-DQ a1a2 chain (ii) a transmembrane domain and wherein the first and/or second polypeptide further comprises an immune receptor intracellular signaling domain; wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQBT1*02, respectively; or wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA 102 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

In one embodiment, the chimeric HLA class || molecule comprises: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ B1B2 chain (ii) a transmembrane domain; and

(iii) an immune receptor intracellular signalling domain; and {b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain (ii) a transmembrane domain; and wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and

B182 chains are encoded by the genes HLA-DQA1*03 and HLA-DQB1*03, respectively.

In one embodiment, the chimeric HLA class II molecule comprises: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ B182 chain; (ii) a transmembrane domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain (ii) a transmembrane domain; and (iii) an immune receptor intracellular signalling domain; and wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA7*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

In one embodiment, the chimeric HLA class || molecule comprises: (a) a first polypeptide comprising: (i) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ B1B2 chain (ii) a transmembrane domain; and (iii) an immune receptor intracellular signalling domain; and (b) a second polypeptide comprising: (i) a second extracellular domain comprising an HLA-DQ a1a2 chain (ii) a transmembrane domain; and (ii) an immune receptor intracellular signalling domain; and wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the genes HLA-DQA1*05 and

The inventors surprisingly discovered that a chimeric HLA class II molecule according to the invention is able to treat and/or prevent celiac disease.

Tissue transglutaminase (TG2) is a crucial factor in celiac disease because it promotes gluten-specific T cell responses. In vivo, TG2 causes selective deamidation of gluten, which in turn, causes the generation of a series of proline-rich and TG2 modified gluten-derived peptides that bind to HLA-DQ2 or -DQ8 molecules with high affinity. Responses to these peptides are detectable in all patients with CD. In particular the T cell response in HLA-DQ2.5 positive CD patients is focussed on 2 overlapping 9 amino acid peptide sequences in the N- terminal part of the a-gliadins (e.g. (i) HLA-DQ2-glia-a1a: PFPQPELPY (SEQ ID NO:3) and

HLA-DQ2-glia-a2: PQPELPYPQ (SEQ ID NO: 4), which are both present in the 11-mer sequence PFPQPELPYPQ, (SEQ ID NO: 5); or (ii) HLA-DQ2-glia-a1b: PYPQPELPY (SEQ ID

NO:69) and HLA-DQ2-glia-a2: PQPELPYPQ (SEQ ID NO: 4), which are both present in the 11-mer sequence PYPQPELPYPQ, (SEQ ID NO: 70)), and homologous sequences thereof in the w-gliadins (PFPQPEQPF (SEQ ID NO: 44) and PQPEQPFPW (SEQ ID NO: 45), combined PFPQPEQPFPW (SEQ ID NO: 46)). Of note, the latter w-gliadin sequences are identical to sequences found in the hordeins of barley and the secalins of rye, providing an explanation for the shared toxicity of wheat, barley and rye for patients with CD. Similarly, in

HLA-DQS8 positive patients the response is directed to an immunodominant peptide sequence from the C-terminal part of the a-gliadins (EGSFQPSQE, SEQ ID NO: 61).

In one embodiment, the one or more gluten-derived epitopes are from the protein gliadin.

Suitably, the gliadin may be alpha gliadin.

Suitably, the one or more alpha gliadin-derived epitope may be glia-a, for example glia-a1 or glia-a2.

Suitably, the one or more alpha gliadin-derived epitope may be glia-a1, for example glia- at or glia-a2. In one embodiment, the one or more alpha gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 3. SEQ ID NO: 3 corresponds to the peptide HLA-

DQ2-glia-a1a. HLA-DQ2-glia-a1a is an immunodominant peptide of celiac disease. Therefore, it is advantageous for a cell (such as a CD8+ T cell) to express a CHAR according to the invention (which presents the immunodominant peptide HLA-DQ2-glia-a1a) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing

CD.

In one embodiment, the one or more alpha gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 69. SEQ ID NO: 69 corresponds to the peptide HLA-DQ2-glia- a1b. HLA-DQ2-glia-a1b is an immunodominant peptide of celiac disease. Therefore, it is advantageous for a cell (such as a CD8+ T cell) to express a CHAR according to the invention (which presents the immunodominant peptide HLA-DQ2-glia-a1b) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing CD.

Suitably, the one or more alpha gliadin-derived epitope may be glia-a2.

In one embodiment, the one or more alpha gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 4. SEQ ID NO:4 encodes HLA-DQ2-glia-a2. HLA-DQ2-glia-a2 is also an immunodominant peptide of celiac disease. Therefore, it is advantageous for a cell (such as a CD8+ T cell) to express a CHAR according to the invention (which presents the immunodominant peptide HLA-DQ2-glia-a2) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing CD.

In one embodiment, the gluten-derived peptide comprises two or more alpha gliadin- derived epitopes. In a preferred embodiment, the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 3 and SEQ ID NO: 4. As it would be appreciated a cell (such as a CD8+ T cell) that expresses such a chimeric HLA class || molecule would be capable of forming two distinct HLA: peptide complexes, each targeting and eliminating a distinct CD4+ T cell population that is responsible causing CD.

In one embodiment, the gluten-derived peptide comprises two or more alpha gliadin- derived epitopes. In a preferred embodiment, the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 69 and SEQ ID NO: 4. As it would be appreciated a cell (such as a CD8+ T cell) that expresses such a chimeric HLA class || molecule would be capable of forming two distinct HLA: peptide complexes, each targeting and eliminating a distinct CD4+ T cell population that is responsible causing CD.

In one embodiment, the gluten-derived peptide comprises the amino acid sequence of

SEQ ID NO: 5. The immunodominant HLA-DQ2-glia-a1a and HLA-DQ2-glia-a2 are both contained within the 11-mer sequence PFPQPELPYPQ (SEQ ID NO:5). Surprisingly, and advantageously, a cell (such as a CD8+ T cell) expressing a CHAR comprising and therefore presenting the 11-mer sequence PFPQPELPYPQ has the ability to target distinct CD4+ T cell populations, directed towards either HLA-DQ2-glia-a1a or HLA-DQ2-glia-a2, simultaneously.

SEQ ID NO: 70. The immunodominant HLA-DQ2-glia-a1b and HLA-DQ2-glia-a2 are both contained within the 11-mer sequence PYPQPELPYPQ (SEQ ID NO:70). Surprisingly, and advantageously, a cell (such as a CD8+ T cell) expressing a CHAR comprising and therefore presenting the 11-mer sequence PyPQPELPYPQ has the ability to target distinct CD4+ T cell populations, directed towards either HLA-DQ2-glia-a1b or HLA-DQ2-glia-a2, simultaneously.

In one embodiment, the one or more alpha gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 61. SEQ ID NO: 61 corresponds to the peptide HLA-DQ8-glia- al. Therefore, it is advantageous for a cell (e.g. a CD8+ T cell) to express a CHAR according to the invention (which presents the immunodominant peptide HLA-DQ8-glia-a1) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing

CD.

In one embodiment, the gluten-derived peptide comprises two or more alpha gliadin- derived epitopes. In a preferred embodiment, the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 61 and one or more of SEQ ID NO: 3, SEQ ID NO: 4, and/or

SEQ ID NO: 5. As it would be appreciated a cell (e.g. a CD8+ T cell) that expresses such a chimeric HLA class II molecule would be capable of forming two distinct peptide:HLA complexes, each targeting and eliminating a distinct CD4+ T cell population that is responsible causing CD.

In one embodiment, the gluten-derived peptide comprises two or more alpha gliadin- derived epitopes. In a preferred embodiment, the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 61 and one or more of SEQ ID NO: 69, SEQ ID NO: 4, and/or

SEQ ID NO: 70. As it would be appreciated a cell (e.g. a CD8+ T cell) that expresses such a chimeric HLA class II molecule would be capable of forming two distinct peptide:HLA complexes, each targeting and eliminating a distinct CD4+ T cell population that is responsible causing CD.

In one embodiment, the gliadin is omega gliadin. In one embodiment, the one or more omega gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 44. SEQ

ID NO: 44 encodes HLA-DQ2-glia-Q1. HLA-DQ2-glia-Q1 is another immunodominant peptide of celiac disease. Therefore, it is advantageous for a cell (e.g. a CD8+ T cell) to express a

CHAR according to the invention (which presents the immunodominant peptide HLA-DQ2- glia-Q1) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing CD.

In one embodiment, the one or more omega gliadin-derived epitope comprises the amino acid sequence of SEQ ID NO: 45. SEQ ID NO:45 encodes HLA-DQ2-glia-Q2. HLA-DQ2-glia-

Q2 is an also an immunodominant peptide of celiac disease. Therefore, it is advantageous for acell (e.g. a CD8+ T cell) to express a CHAR according to the invention (which presents the immunodominant peptide HLA-DQ2-glia-Q2) because it has the ability to be effective at killing a key population of CD4+ T cells responsible for causing CD.

In one embodiment, the gluten-derived peptide comprises two or more omega gliadin- derived epitopes. In a preferred embodiment, the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 44 and SEQ ID NO: 45. As it would be appreciated a cell (e.g. a CD8+ T cell) that expresses such a chimeric HLA class Il molecule would be capable of forming a two distinct peptide:HLA complexes, each targeting and eliminating a distinct CD4+

T cell population that is responsible causing CD.

SEQ ID NO: 46. The immunodominant HLA-DQ2-glia-Q1 and HLA-DQ2-glia-Q2 are both contained within the 11-mer sequence (SEQ ID NO: 48). Surprisingly, and advantageously, a cell (such as a CD8+ T cell) expressing a CHAR comprising and therefore presenting the 11-

mer sequence PFPQPEQPFPW has the ability to target distinct CD4+ T cell populations, directed towards either HLA-DQ2-glia-Q1 or HLA-DQ2-glia-Q2, simultaneously.

In one embodiment, the gluten-derived peptide comprises two or more gluten-derived epitopes, e.g. gliadin-derived epitopes. Suitably, in an embodiment that comprises two or more gluten-derived epitopes, at least one of the epitopes may be an alpha gliadin-derived epitope, and/or at least one of the epitopes may be an omega gliadin-derived epitope. In one embodiment, the two or more gluten-derived epitopes may be selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 69, SEQ ID NO: 70,

SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 48. Suitably, when the gluten-derived peptide comprises two or more gluten-derived epitopes, one epitope may be according to SEQ

ID NO: 5, and one may be according to SEQ ID NO: 46. Suitably, when the gluten-derived peptide comprises two or more gluten-derived epitopes, one epitope may be according to SEQ

ID NO: 70, and one may be according to SEQ ID NO: 46. Advantageously, such a CHAR comprising two or more distinct gluten-derived epitopes is able to target and eliminate two or more distinct CD4+ T cell populations that are responsible for CD.

In one embodiment, the gluten-derived peptide comprises four or more gluten-derived epitopes, e.g. gliadin-derived epitopes. Suitably, in an embodiment that comprises four or more gluten-derived epitopes, at least one of the epitopes may be an alpha gliadin-derived epitope, and/or at least one of the epitopes may be an omega gliadin-derived epitope. In one embodiment, the four or more gluten-derived epitopes may be selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 69, SEQ ID NO: 70,

SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 48. Suitably, when the gluten-derived peptide comprises four or more gluten-derived epitopes, the four or more may be SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 44, and SEQ ID NO: 45. Suitably, when the gluten-derived peptide comprises four or more gluten-derived epitopes, the four or more may be SEQ ID NO: 69, SEQ ID NO: 4, SEQ ID NO: 44, and SEQ ID NO: 45. Advantageously, such a CHAR comprising two or more distinct gluten-derived epitopes is able to target and eliminate four or more distinct CD4+ T cell populations that are responsible for CD.

In some embodiments, wherein the gluten-derived peptide comprises two or more gluten- derived epitopes, the gluten-derived peptide comprises a linker separating the amino acid sequences of the epitopes.

In some embodiments, the first extracellular domain further comprises a linker sequence between the gluten-derived peptide and the HLA-DQ B1B2 chain. An example of a suitable linker is provided as SEQ ID NO: 6.

Peptides that are longer than the conventional 9mer sequence presented by HLA may be used within the chimeric HLA class Il molecule or fusion polypeptide provided herein. For example, the gluten-derived peptides described herein may be from 9 to 35 amino acids long.

For the avoidance of doubt, in this context, the peptides can have a total of 9 to 35 amino acids, which includes the one or more gluten-derived epitope. The additional amino acids may be located N-terminal or C-terminal to the one or more gluten-derived epitope sequence.

Alternatively, the additional amino acids may flank the one or more gluten-derived epitope sequence (i.e. such that there are additional amino acid(s} N-terminal and C-terminal of the one or more gluten-derived epitope sequence). Additional amino acids located N-terminal, C- terminal or flanking the one or more gluten-derived epitope are referred to collectively as “flanking amino acids” herein.

The N-terminus of a peptide (also known as the amino-terminus, N-terminus, N-terminal end or amine-terminus) is the start of a peptide terminated by an amino acid with a free amine group (-NH2). By convention, peptide sequences are written N-terminus to C-terminus (from left to right). The C-terminus (also known as the carboxyl-terminus, carboxy-terminus, C- terminal tail, C-terminal end, or COOH-terminus) is the end of an amino acid chain (protein or polypeptide), terminated by a free carboxyl group (-COOH).

As used herein, the terms “N-terminal” and “C-terminal” are used to describe the relative position of e.g. a sequence within a peptide or polypeptide. Accordingly, a sequence that is “N-terminal” is positioned closer (in relative terms) to the N-terminus than to the C-terminus of the peptide or polypeptide. Conversely, a domain that is “C-terminal” is positioned (in relative terms) closer to the C- terminus than to the N-terminus of the peptide or polypeptide. As used herein, the term “positioned” refers to the location of the sequence within the linear amino acid sequence of the peptide or polypeptide.

Where the peptides described herein include additional amino acids located N- terminal,

C-terminal or flanking the one or more gluten-derived epitopes, any appropriate additional amino acid sequences may be included. For example, the additional amino acids may be amino acid sequences that are naturally located N-terminal, C-terminal or flanking the gluten- derived epitope sequence. In a particular example, the additional amino acids are located at

N- terminal to the one or more gluten-derived epitope sequence and may be the natural sequence that is found N- terminal to the one or more gluten-derived epitope. In another example, the additional amino acids may be all be located C-terminal to the one or more gluten-derived epitope sequence and may be the natural sequence that is found C-terminal to the one or more gluten-derived epitope sequence. Alternatively, the additional amino acids may flank the one or more gluten-derived epitope sequence (i.e. such that there are additional amino acid(s) N-terminal and C-terminal of the one or more gluten-derived epitope sequence) and may be the natural sequence that flank the one or more gluten-derived epitope sequence.

Advantageously, flanking amino acids mimic natural peptide:HLA complex formation, resulting in higher binding affinity between the peptide:HLA complex and CD4 T cells.

In one example, when the gluten-derived peptide comprises the gluten-derived epitope of

SEQ ID NO: 5, the gluten-derived peptide may comprise an N-terminal flanking sequence of

QLQ and/or a C-terminal flacking region of PQL. In other words, the gluten-derived peptide may comprise the sequence QLQPFPQPELPYPQ (SEQ ID NO: 83),

QLQPFPQPELPYPQPQL (SEQ ID NO: 64) or PFPQPELPYPQPQL (SEQ ID NO: 65).

SEQ ID NO: 48, the gluten-derived peptide may comprise an N-terminal flanking sequence of PQQ and/or a C-terminal flacking region of QPQ. In other words, the gluten-derived peptide may comprise the sequence PQQPFPQPEQPFPW (SEQ ID NO: 66),

PQQPFPQPEQPFPWQPQ (SEQ ID NO: 67) or PFPQPEQPFPWQPQ (SEQ ID NO: 68).

SEQ ID NO: 70, the gluten-derived peptide may comprise an N-terminal flanking sequence of QLQ and/or a C-terminal flacking region of PQL. In other words, the gluten-derived peptide may comprise the sequence QLQPYPQPELPYPQ (SEQ ID NO: 71),

QLQPYPQPELPYPQPQL (SEQ ID NO: 72) or PYPQPELPYPQPQL (SEQ ID NO: 73).

The invention also provides a fusion polypeptide, comprising, in an N-terminal to C- terminal orientation: (a) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ B1B2 chain; (b) a transmembrane domain; (c) a peptide cleavage signal; (d) a second extracellular domain comprising an HLA-DQ a1a2 chain; {e) a transmembrane domain; wherein the polypeptide further comprises an immune receptor intracellular signalling domain located at the C-terminus of (b) and/or (e), and wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA7*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

B1B2 chains are encoded by the gene HLA-DQA1*03 and HLA-DQB1*03, respectively.

For avoidance of doubt the fusion polypeptide described herein may form a chimeric HLA class Il molecule as described herein. For example, the fusion polypeptide may be cleaved at the peptide cleavage signal to form a first polypeptide (comprising an a1a2 chain) and a second polypeptide (comprising a B1B2 chain), wherein the first and second polypeptide together form a chimeric HLA class II molecule of the invention. Several fusion polypeptides are described herein, see for example SEQ ID NO: 1, SEQ ID NO: 43 and SEQ ID NO: 55, which are described in the examples section below).

As will be appreciated, embodiments described in relation to the chimeric HLA class II molecule of the invention apply equally to the fusion polypeptide of the invention unless the context specifically indicates otherwise.

In one embodiment the fusion polypeptide comprises in an N-terminal to C-terminal orientation: (a) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8182 chain; (b1) a transmembrane domain; (b2) an immune receptor intracellular signalling domain (c) a peptide cleavage signal; (d) a second extracellular domain comprising an HLA-DQ a1a2 chain; (e) a transmembrane domain; and wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and

B1B2 chains are encoded by the gene HLA-DQA1*03 and HLA-DQB1*03, respectively.

In another embodiment the fusion polypeptide comprises in an N-terminal to C-terminal orientation: (a) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8182 chain; (b) a transmembrane domain; (c) a peptide cleavage signal; (d) a second extracellular domain comprising an HLA-DQ a1a2 chain; (e1) a transmembrane domain; (e2) an immune receptor intracellular signalling domain; and wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*05 and

B1B2 chains are encoded by the gene HLA-DQA1*03 and HLA-DQB1*03, respectively.

In another embodiment the fusion polypeptide comprises in an N-terminal to C-terminal orientation: (a) a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8182 chain; (b1) a transmembrane domain; (b2) an immune receptor intracellular signalling domain (c) a peptide cleavage signal; (d) a second extracellular domain comprising an HLA-DQ a1a2 chain;

(e1) a transmembrane domain; (e2) an immune receptor intracellular signalling domain; and wherein the HLA-DQ a1a2 and B1B2 chains are encoded by the gene HLA-DQA1*05 and

HLA-DQB1*02, respectively; or wherein the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*02 and HLA-DQB1*02, respectively; or wherein the HLA-DQ a102 and

B1B2 chains are encoded by the gene HLA-DQA1*03 and HLA-DQB1*03, respectively.

In one embodiment, the fusion polypeptide may further comprise a linker sequence between the immune receptor intracellular signalling domain and the peptide cleavage signal.

In one embodiment, the polypeptide cleavage signal is a viral self-cleaving polypeptide. In a preferred embodiment, the peptide cleavage signal is a viral self-cleaving 2A polypeptide (see for example SEQ ID NO: 11). Other exemplary protease cleavage sites include, but are not limited to, the cleavage sites of potyvirus Ma proteases (e.g., tobacco etch virus protease), potyvirus HC proteases, potyvirus P1 (P35) proteases, byovirus Ma proteases, byovirus RNA- 2-encoded proteases, aphthovirus L proteases, enterovirus 2A proteases, rhinovirus 2A proteases, picorna 3C proteases, comovirus 24K proteases, nepovirus 24K proteases, RTSV (rice tungro spherical virus) 3C-like protease, PYVF (parsnip yellow fleck virus) 3C-like protease, heparin, thrombin, factor Xa and enterokinase. In one embodiment, the polypeptide cleavage signal may be an internal ribosome entry site (IRES) sequence.

In one embodiment, the fusion polypeptide may further comprise an internal ribosome entry site (IRES) after the transmembrane domain that is C-terminal to the a1a2 chain. In another embodiment, the fusion polypeptide further comprises a IRES and truncated nerve growth factor receptor (NGFR) at the C-terminus of the fusion polypeptide (in an N- to C- terminal orientation).

As described herein, the chimeric HLA class || molecule and the fusion polypeptide of the invention comprise HLA-DQ a1a2 and B1B2 chains. As would be clear to a person of skill in the art, the HLA-DQ a1a2 and 8182 chains within a chimeric HLA class Il molecule or fusion polypeptide must be capable of forming a heterodimeric HLA class Il complex. In a suitable embodiment, the HLA-DQ a1a2 and B1B2 chains are encoded by the gene HLA-DQA1*05 and HLA-DQB1*02, respectively; or the HLA-DQ a102 and B1B2 chains are encoded by the gene HLA-DQA1*02 and HLA-DQB1*02, respectively; or the HLA-DQ a1a2 and 8182 chains are encoded by the gene HLA-DQA1*03 and HLA-DQB1*03, respectively.

In an embodiment when the HLA-DQ a1a2 and 8182 chains are encoded by the gene

HLA-DQA1*02 and HLA-DQB1*02, HLA-DQA1*02 may be HLA-DQA1*0201 or HLA-

DQA1*0202.

In an embodiment when the HLA-DQ a1a2 and 8182 chains are encoded by the gene

HLA-DQA1*03 and HLA-DQB1*03, HLA-DQA1*03 may be HLA-DQA1*0301 or HLA-

DQA1*0302, and optionally HLA-DQB1*03 may be HLA-DQB1*0302.

As described elsewhere herein, the chimeric HLA class II molecule or the fusion polypeptide of the invention comprise a transmembrane domain. In one embodiment, the transmembrane domain is selected from the group consisting of: alpha or beta chain of CD28,

CD4, CD5, CD8, CD9, CD16, CD22, CD27, CD33, CD37, CD45, CD64, CD80, CD86, CD134,

CD137, CD152, CD154, PD1, HLA-DR, HLA-DQ or HLA-DP. In a specific embodiment, the transmembrane domain is a CD28 transmembrane domain (see for example, SEQ ID NO:8).

Suitably, the transmembrane domain is a HLA-DQ transmembrane domain.

As described elsewhere herein, the chimeric HLA class II molecule or the fusion polypeptide of the invention comprise a immune receptor intracellular signalling domain. In one embodiment, the immune receptor intracellular signalling domain comprises one or more co-stimulatory signalling domains. In a specific embodiment, the one or more co-stimulatory signalling domain is selected from the group consisting of: TLR1, TLR2, TLR3, TLR4, TLRS,

TLR6, TLR7, TLRS, TLR9, TLR10, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (0X40), CD137 (4-1BB), CD154 (CD40L), CD278 (ICOS), DAP10,

LAT, NKD2C, SLP76, TRIM, and ZAP70 co-stimulatory signalling domain. For example, the one or more co-stimulatory signalling domain may be a CD28 co-stimulatory signalling domain (see for example SEQ ID NO: 9). Suitably, the one or more co-stimulatory signalling domain may be a CD137 (4-1BB) co-stimulatory signalling domain. Suitably, the one or more co- stimulatory domains may comprise a CD28 co-stimulatory signalling domain and a CD137 co- stimulatory domain.

The immune receptor intracellular signalling domain may (further) comprise a primary signalling domain. In one embodiment, the primary signalling domain may be selected from the group consisting of: FcRy, FCRB, CD3y, CD30, CD3¢, CD3Z, CD22, CD79b, and CD66d.

In a specific embodiment, the primary signalling domain is CD3( (see for example, SEQ ID

NO: 10).

In one embodiment, the chimeric HLA class II molecule or the fusion polypeptide of the invention may comprise one or more immune receptor signalling domains, e.g. one or more co-stimulatory signalling domains and/or one or more primary signalling domains.

The invention provides a nucleic acid encoding a chimeric HLA class II molecule according to the invention, or a fusion polypeptide according to the invention. Suitably, the nucleic acid may be DNA or RNA, or a combination thereof.

The invention provides a vector comprising said nucleic acid. Any appropriate vector can be used. By way of example only, the vector may be a plasmid, a cosmid, or a viral vector, such as a retroviral vector or a lentiviral vector. Adenovirus, adeno-associated virus, vaccinia virus, canary poxvirus, herpes virus, minicircle vectors and naked (synthetic) DNA/RNA may also be used (for details on minicircle vectors, see for example non-viral Sleeping Beauty transposition from minicircle vectors as published by R Monjezi et al. Leukemia 2017).

Alternatively, single stranded or double stranded DNA or RNA can be used to transfect lymphocytes with a TCR of interest (see Roth ef al 2018 Nature vol 559; page 405).

In one example, the vector is a plasmid, a viral vector, or a cosmid, optionally wherein the vector is selected from the group consisting of a retrovirus, lentivirus, adeno-associated virus, adenovirus, vaccinia virus, canary poxvirus, herpes virus, minicircle vector and synthetic

DNA or RNA.

Preferably the (expression) vector is capable of propagation in a host cell and is stably transmitted to future generations.

The vector may comprise regulatory sequences. "Regulatory sequences” as used herein, refers to, DNA or RNA elements that are capable of controlling gene expression.

Examples of expression control sequences include promoters, enhancers, silencers, TATA- boxes, internal ribosomal entry sites (IRES), attachment sites for transcription factors, transcriptional terminators, polyadenylation sites etc. Optionally, the vector includes one or more regulatory sequences operatively linked to the nucleic acid sequence to be expressed.

Regulatory sequences include those which direct constitutive expression, as well as tissue- specific regulatory and/or inducible sequences.

Optionally, the vector comprises the nucleic acid sequence of interest operably linked to a promoter. "Promoter", as used herein, refers to the nucleotide sequences in DNA to which

RNA polymerase binds to start transcription. The promoter may be inducible or constitutively expressed. Alternatively, the promoter is under the control of a repressor or stimulatory protein. The promoter may be one that is not naturally found in the host cell (e.g. it may be an exogenous promoter). The skilled person in the art is well aware of appropriate promoters for use in the expression of target proteins, wherein the selected promoter will depend on the host cell. "Operably linked" refers to a single or a combination of the below-described control elements together with a coding sequence in a functional relationship with one another, for example, in a linked relationship so as to direct expression of the coding sequence.

The vector may comprise a transcriptional terminator. “Transcriptional terminator” as used herein, refers to a DNA element, which terminates the function of RNA polymerases responsible for transcribing DNA into RNA. Preferred transcriptional terminators are characterized by a run of T residues preceded by a GC rich dyad symmetrical region.

The vector may comprise a translational control element. “Translational control element”, as used herein, refers to DNA or RNA elements that control the translation of mRNA.

Preferred translational control elements are ribosome binding sites. Preferably, the translational control element is from a homologous system as the promoter, for example a promoter and its associated ribozyme binding site. Preferred ribosome binding sites are known, and will depend on the chosen host cell.

The vector may comprise restriction enzyme recognition sites. "Restriction enzyme recognition site" as used herein, refers to a motif on the DNA recognized by a restriction enzyme.

Preferably the vector comprises those genetic elements which are necessary for expression of the binding proteins described herein by a host cell. The elements required for transcription and translation in the host cell include a promoter, a coding region for the protein(s) of interest, and a transcriptional terminator.

A person of skill in the art will be well aware of the molecular techniques available for the preparation of (expression) vectors and how the (expression) vectors may be transduced or transfected into an appropriate host cell (thereby generating a modified cell described further below). The (expression) vector system described herein can be introduced into cells by conventional techniques such as transformation, transfection or transduction. “Transformation”, “transfection” and “transduction” refer generally to techniques for introducing foreign (exogenous) nucleic acid sequences into a hast cell, and therefore encompass methods such as electroporation, microinjection, gene gun delivery, transduction with retroviral, lentiviral or adeno-associated vectors, lipofection, superfection etc. The specific method used typically depends on both the type of vector and the cell. Appropriate methods for introducing nucleic acid sequences and vectors into host cells such as human cells are well known in the art; see for example Sambrook et al (1989) Molecular Cloning, A Laboratory

Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y; Ausubel et al (1987) Current

Protocols in Molecular Biology, John Wiley and Sons, Inc., NY; Cohen et al (1972) Proc. Natl.

Acad. Sci. USA 69, 2110; Luchansky et al (1988) Mol. Microbiol. 2, 637-846. Further conventional methods that are suitable for preparing expression vectors and introducing them into appropriate host cells are described in detail in WO2016/071758 for example.

The invention provides a cell comprising one or more nucleic acid, vector, chimeric HLA class Il molecule, and/or fusion polypeptide according to the invention.

The invention also provides a cell comprising at least two of the following: a) a first nucleic acid, vector, chimeric HLA class || molecule, or fusion polypeptide according to any one of the preceding claims, wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*05 and HLA-DQB1*02, respectively; b) a second nucleic acid, vector, chimeric HLA class Il molecule, or fusion polypeptide according to any one of the preceding claims, wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA7*02 and HLA-DQB1*02, respectively; and/or ¢) a third nucleic acid, vector, chimeric HLA class II molecule, or fusion polypeptide according to any one of the preceding claims, wherein the HLA-DQ a1a2 and 8182 chains are encoded by the genes HLA-DQA1*03 and HLA-DQB1*03, respectively.

Such a cell is advantageous as it is capable of simultaneously targeting distinct T cell populations that recognise distinct HLA-DQ2:peptide and/or HLA-DQ8: peptide complexes.

In one example, the cell of the invention may encode two CHARS separated by a linker (e.g. one that encodes HLA-DQ2 and PFPQPELPYPQ; linked to another that encodes HLA-

DQ8 and EGSFQPSQE) such that at least two distinct CHARS are expressed on the same T cell (each one targeting a different TCR repertoire). This would be useful in patients that are both HLA-DQ2 positive (95% of patients) and HLA-DQ8 positive, as the resultant CHAR CD8+

T cell would be able to target TCRs that recognise: HLA-DQ2:PFPQPELPY complexes; HLA-

DQ2:PQPELPYPQ complexes and HLA-DQ8:EGSFQPSQE complexes.

In one example, the cell of the invention may encode two CHARS separated by a linker (e.g. one that encodes HLA-DQ2 and PYPQPELPYPQ; linked to another that encodes HLA-

DQ8 and EGSFQPSQE) such that at least two distinct CHARSs are expressed on the same T cell (each one targeting a different TCR repertoire). This would be useful in patients that are both HLA-DQ2 positive (95% of patients) and HLA-DQ8 positive, as the resultant CHAR CD8+

T cell would be able to target TCRs that recognise: HLA-DQ2:PYPQPELPY complexes; HLA-

DQ2:PQPELPYPQ complexes and HLA-DQ8:EGSFQPSQE complexes.

In one example, the cell of the invention may encode two CHARS separated by a linker (e.g. one that encodes HLA-DQ2 and PFPQPEQPFPW, linked to another that encodes HLA-

T cell would be able to target TCRs that recognise: HLA-DQ2: PFPQPEQPF complexes; HLA-

DQ2: PQPEQPFPW complexes and HLA-DQ8:EGSFQPSQE complexes.

The cell is typically a eukaryotic cell, and particularly a human cell. Suitably, the cell may be a human immune cell, for example a T cell, NK cell, or an innate lymphoid cell (ILC).

Suitably, the T cell may be selected from the group consisting of CD8* T cell, CD3+ T cell,

NK T cell, and gamma delta T cell (yd T cell), or a mixture of any one thereof. Suitably, the cell is a CD8* T cell. Suitably, the cell is a CD3+ T cell. The cell may be an autologous or allogeneic cell. “Allogeneic cell” refers to a cell derived from a different individual to the individual to which it is later administered. In other words, the cell may be an isolated cell from a distinct individual compared to the subject to be treated. “Autologous cell” refers to a cell derived from the individual to which it is also later administered. In other words, the cell may be an isolated cell from the subject that is to be treated.

The invention provides a composition comprising a nucleic acid, vector chimeric HLA class Il molecule, fusion polypeptide and/or cell according to the invention.

The invention provides a pharmaceutical composition comprising a nucleic acid, vector, chimeric HLA class II molecule, fusion polypeptide and/or cell according to the invention.

Suitably, said pharmaceutical composition is for use as a medicament.

The invention provides a pharmaceutical composition comprising a nucleic acid, vector, chimeric HLA class Il molecule, fusion polypeptide and/or cell according to the invention for use in treating or preventing celiac disease in an HLA-DQ positive subject.

CHAR, fusion polypeptides, compositions, nucleic acid molecule, or cells of the invention may advantageously be used to treat or prevent celiac disease in a subject. An appropriate composition may be selected independently of the HLA serotype of the subject.

In the alternative, an appropriate composition may be selected on the basis of the HLA-DQ serotype and/or HLA-DR serotype of the subject.

Suitably, the subject is HLA-DQA1*02-HLA-DQB1*02, HLA-DQA1*05-HLA-

DQB1*02, and/or HLA-DQA1*03-DQB1*03 positive.

Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of them mean “including but not limited to”, and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps.

Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith.

Various aspects of the invention are described in further detail below.

Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein find use in the practice of the present invention, the preferred methods and materials are described herein. Accordingly, the terms defined immediately below are more fully described by reference to the Specification as a whole. Also, as used herein, the singular terms "a", "an," and "the" include the plural reference unless the context clearly indicates otherwise. Unless otherwise indicated, nucleic acids are written left to right in &' to 3 orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively. It is to be understood that this invention is not limited to the particular methodology, protocols, and reagents described, as these may vary, depending upon the context they are used by those of skill in the art.

Aspects of the invention are demonstrated by the following non-limiting examples.

Examples

The inventors have investigated whether cytotoxic CD8 T cells expressing HLA-DQ2 coupled to a costimulatory signalling and CD34 signalling domain and loaded with the relevant immunodominant a- and w-gliadin peptides would be able to specifically interact with gliadin- specific T cells from patients, resulting in the elimination of these disease-causative CD4 T cells. To obtain proof of concept they generated the chimeric HLA class II molecule constructs (also referred to as chimeric HLA antigen receptor (CHAR) constructs) shown in Figure 1.

The inventors identified that the immunodominant HLA-DQ2-glia-a1a and HLA-DQ2- glia-a2 are both contained within the 11-mer sequence PFPQPELPYPQ. They reasoned that if the 11-mer sequence PFPQPELPYPQ was encoded within the HLA-DQ2 beta construct both the HLA-DQ2-glia-a1a and HLA-DQ2-glia-a2 epitope would be expressed by the CHAR

T cells, allowing the ability to target CD4 T cells directed towards both HLA-DQ2-glia-a1a and

HLA-DQ2-glia-a2 simultaneously with a single CAR T cell. As a control, HLA-DQ2 constructs containing the alternative CLIP peptide were also generated.

Cells were transduced with the HLA-DQ2 CHAR constructs by using retroviral transduction. Expression of the HLA-DQ2 CHAR construct was first confirmed on K562 cells using different human monoclonal antibodies specific for HLA-DQ2. K562 cells expressing the

HLA-DQ CHAR constructs were then enriched and used as stimulator cells to confirm that the

DQ-glia-a1a and DQ-glia-a2 epitopes were presented in HLA-DQ2 on the cell surface. This was investigated using a panel of CD patient-derived T cell clones described previously. In short, those T cell clones were derived from small intestinal biopsies obtained from CD patients undergoing endoscopy for diagnostic purposes. Their (fine)specificity was determined by testing against synthetic variants of gluten-derived peptides, TCR sequences were determined where appropriate, and structural studies underpinned the observed specificity.

The inventors tested three T cell clones, one specific for the HLA-DQ2-glia-a1a epitope (S82), one specific for the HLA-DQ2-glia-a2 epitope (S16), and one specific for the unrelated

HLA-DQ2-glia-y2 epitope (SV30). To determine the conformation and integrity of the HLA-

DQ2 CHAR construct, the three T cell clones (specific for each of S2, S16 and SV30) were incubated with CHAR construct transduced K562 cells and it was tested whether the T cell clones recognized the CHAR construct. The inventors observed IL-2 and IFNy production when the HLA-DQ2-glia-a1a and HLA-DQ2-glia-a2 specific T cell clones were incubated with

K562 cells transduced with gluten-derived peptide-linked HLA-DQ2 CHAR construct, but not when incubated with K562 cells transduced with CLIP peptide -linked HLA-DQ2 CHAR construct. Moreover, the T cell clone recognizing the unrelated HLA-DQ2-glia-y2 epitope did not respond to K562 cells transduced with either CHAR constructs confirming the correct conformation and integrity of the HLA-DQ2- ala and DQ-glia-a2 epitope in the HLA-DQ2

CHAR construct (Figure 2).

With the cell surface expression, correct conformation, and integrity of the HLA-DQ2

CHAR construct shown, the ability of CD8+ HLA-DQ2 CHAR T cells to specifically lyse gluten- specific T cell clones was determined (Figure 3). For this purpose, CD8+ T cells were transduced with the HLA-DQ2 CHAR constructs. To investigate if the generated CHAR T cells have the capacity to specifically eliminate T cells specific for the HLA-DQ2-glia-a1a and HLA-

DQ2-glia-a2 epitopes, CD patient-derived T cell clones S2 (glia- a1), S16 (glia-a2) and SV30 (glia-y2) were used. Next, the CD8 T cells of two different healthy donors transduced with either DQ2-glia-a CHAR (SEQ ID NO:1), DQ2-CLIP CHAR, or mock were incubated for 6 hours with 51Cr labelled T cell clones (S2, S16 or SV30). As demonstrated in Figure 3, the

DQ2 glia-a CHAR T cells of both donors have the capacity to specifically eliminate CD4 T cells specific for the HLA-DQ2-glia-a1a (S2) and HLA-DQ2-glia-a2 epitope (S16), in a dose- dependent manner, whereas the control T cell clone (SV30) was not killed, demonstrating that a single CHAR T cell can eliminate patient-derived T cells specific for the immunodominant

HLA-DQ2-glia-a1a and HLA-DQ2-glia-a2 epitopes simultaneously.

Construction design and retroviral production

Retroviral vectors encoding CHAR molecules included the signal peptide, followed by the sequence of the overlapping gliadin-alpha-1 and gliadin-alpha-2 peptides (CCGTTTCCGCAGCCGGAACTGCCGTACCCGCAG — SEQ ID NO:31) with additional flanking amino acids, which was connected via a linker to the extracellular part of an HLA-

DQB1*02:01 coupled to the transmembrane and intracellular region of CD28, and intracellular signaling domains of CD3Z, coupled to P2A allowing splicing. Subsequently, the extracellular

HLA-DQA1*05:01 molecule is followed by the CD28 transmembrane and intracellular region, as well as the CD34 intracellular signaling domains. This is coupled with an internal ribosome entry site (IRES) that permits co-expression of truncated nerve growth factor receptor (NGFR).

For the purpose of control, a CHAR construct was engineered with the sequence of the alternative CLIP peptide (CCGCTGCTGATGCAGGCGCTGCCGATG — SEQ ID NO:62) with additional flanking amino acids, instead of the gliadin-alpha peptide. Alternatively, a vector containing only NGFR was used as a mock. Phoenix-AMPHO (ATCC, CRL-3213) or Phoenix-

GALV [1] were transiently transfected with various constructs using Fugene HD Transfection

Reagent (Promega). The retroviral supernatants were collected and stored at -80°C after 48 hours.

Generation of CHAR-expressing cells 24-well flat-bottom culture plates (Greiner Bio-One) were coated with 30 pg/mL of retronectin (Takara) and then blocked with 2% HSA (Sanquin) prior to retroviral transduction.

Retroviral supernatants were added and centrifuged at 3000 g for 20 minutes at 4°C. Following the removal of the retroviral supernatant, the cells were transferred to the virus-coated wells.

After an overnight incubation, the cells were transferred to 24-well flat-bottom plates (Costar).

Seven days after being stimulated by irradiated autologous feeders at 35 Gy and supplemented with phytohaemagglutinin (PHA, Oxoid Microbiology Products, Thermo Fisher

Scientific), the transduced cells were MACS enriched for the NGFR gene marker using an

NGFR-APC antibody (Sanbio, clone ME20.4), and anti-APC MicroBeads (Milteny, clone

BW135/80).

Human CD8 T-cell isolation and cell culture

HLA-DQB1*02:01/HLA-DQA1*05:01 (HLA-DQ2.5) negative healthy donors were selected from the biobank belonging to the Department of Hematology at Leiden University

Medical Center (HEM 008/SH/sh). PBMCs were isolated through standard Ficoll Isopaque separation and preserved through cryopreservation.

PBMCs were thawed and enriched for CD8+ T cells through positive selection using

CD8 Microbeads (Miltenyi). CD8+ T cells were stimulated using autologous feeder cells irradiated at 35 Gy and supplemented with phytohaemagglutinin (PHA, Oxoid Microbiology

Products, Thermo Fisher Scientific). T cells were cultured in T cell medium (TCM) consisting of IMDM (Gibco) with 5% heat-inactivated FBS (Lonza), 5% heat-inactivated human serum (ABOS, Sanquin), 100 U/mL penicillin, 100 pg/mL streptavidin, 2.7mM L-glutamine (Lonza), and 100 IU/mL IL-2 (Chiron).

Gluten-specific T cell culture

Gluten-specific CD4+ T-cell clones S2 and S16 recognize DQ2.5-glia-a1a and DQ2.5- glia-a2 respectively [2]. As it recognizes DQ2.5-y2, SV30 was used as a control [3]. The T-cell clones were thawed and stimulated using allogeneic feeder cells irradiated at 35 Gy, supplemented with PHA and cultured in TCM.

Cell lines and cell culture

K562 (ATCC, CCL-243™) cells were cultured in IMDM with the addition of 10% heat- inactivated FBS, 100 U/mL penicillin, 100 pg/mL streptavidin, 2.7mM L-glutamine (all from

Lonza). All cell lines were regularly tested for mycoplasma contamination.

Flow cytometry

Cells were washed in PBS supplemented with 1% human serum albumin (HSA) and stained with fluorochrome-conjugated antibodies at 4°C for 20-30 minutes using standard flow cytometry protocols.

The CD8 isolated fractions were assessed for purity with PE-labelled CD8B (Beckman Coulter, clone 25T8.5H7) and FITC-coupled CD4 (BD/Pharmingen, clone RPA-

T4). The cell surface expression of CHAR molecules was quantified with human monoclonal antibodies specific for HLA-DQB*02:01 (LB_DQB0201_A, LB_DQB0201_B, and

LB_DQB0201_C), and specific for HLA-DQB*03:03 (LB_DQB0303_A) and secondary PE- conjugated goat-anti-human IgG (Jackson, clone 109-116-098). APC-conjugated NGFR (Sanbio, clone ME20.4) was utilized to establish the transduction efficiency and purity of

CHAR-transduced cells. Cells were washed and fixed in 1% paraformaldehyde before acquisition using either LSRII or Fortessa flow cytometer instruments (BD) and were analyzed using FlowJo software (Tree star). Appropriate controls were included to authenticate antibody specificity.

For the HLA-specific staining, K562 and CD8 CHAR cells were gated on NGFR+.

IFNy and IL2 ELISA

IL2 and IFNy secretion levels were quantified through ELISA using the Invitrogen and

Diaclone kits respectively. To quantify cytokine production, supernatants were collected following overnight cocultures of CD4 T cell clones and K562 CHAR-transduced cells and diluted to 1:5 and 1:125. High-binding plates were coated with IFNy coating antibody overnight to measure IFNy secretion. The plates were then blocked with 10% bovine serum albumin (BSA) for 2 hours at room temperature. Afterward, supernatant and biotinylated detection antibodies were added for 2 hours at room temperature. Then, streptavidin labeled HRP was added for 30 minutes at room temperature. Finally, a substrate containing 6 mg/mL

Tetramethylbenzidine (TMB), and 3% H202 was added. Finally, the reaction was halted with the addition of 2M H2S04.

For the production of IL2 by the CD4+ T cell clones, high-binding plates were coated with an IL2 capture antibody overnight. The plates were subsequently blocked with an

ELISA/ELISPOT diluent for 1 hour at room temperature. Next, the supernatant was then added for 2 hours at room temperature, followed by the addition of an IL2 detection antibody for 1 hour at room temperature. Then, avidin labeled with HRP was added for 30 minutes at room temperature. Finally, the above-mentioned TMB solution was added, and the reaction was terminated with 2M H2S0O4. The absorbance was measured at 450 nm using a microplate reader (Thermo Electron) for both ELISAs.

Chromium release assay

Cytotoxicity was determined through the use of 51-chromium (51Cr) release assays.

CD4+ T cell clones were labeled with 100 pCi 51Cr for one hour at 37 °C. After washing, the cells were cocultured in triplicate with DQ2-glia CHAR, DQ2-CLIP CHAR, or mock transduced

T cells at various E: T ratios (ranging from 9:1 to 0,3:1). Spontaneous 51Cr release of the target cells was measured in culture medium alone, and maximum 51Cr release was determined by the addition of 1% Triton-X100 (Sigma-Aldrich). Supernatants were collected after six hours and then transferred onto Lumaplates (Perkin Elmer). The quantity of released 51Cr was measured on a Microbeta counter (Perkin Elmer). The percentage of specific lysis was calculated using the formula ((experimental 51Cr release — average spontaneous 51Cr release) / (average maximal 51Cr release — average spontaneous 51Cr release)) x 100.

The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.

All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive.

Each feature disclosed in this specification (including any accompanying claims, abstract and drawings), may be replaced by alternative features serving the same, equivalent, or similar purpose, unless expressly stated otherwise. Thus, unless expressly stated otherwise, each feature disclosed is one example only of a generic series of equivalent or similar features.

The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

SEQUENCES

Table 1: CHAR HLA-DQ2.5-glia-ala-glia-a2 molecule

SEQID | CHAR Amino Acid (AA), Sequence

NO POLYPEPTIDE Nucleotide (NT) or

Nucleotide Codon

Optimised {NT OPT} 1 CHAR_HLA- AA 867 MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGQLQP

DQ2.5-glia- FPQPELPYPQPQLGSGSGSLGSGSGSGSGSRDSPEDF ala-glia-a2 VYQFKGMCYFTNGTERVRLVSRSIYNREEIVRFDSDVG molecule (a- EFRAVTLLGLPAAEYWNSQKDILERKRAAVDRVCRHN gliadin) YOLELRTTLQRRVEPTVTISPSRTEALNHHNLLVCSVTD

FYPAQIKVRWFRNDQEETAGVVSTPLIRNGDWTFQIL

VMLEMTPQRGDVYTCHVEHPSLQSPITVEWRAQSES

AQSKFWVLVVVGGVLACYSLLVTVAFHFWVRSKRSRL

LHSDYMNMTPRRPGPTRKHYOQPYAPPRDFAAYRSRV

KFSRSADAPAYQOQGONOQLYNELNLGRREEYDVLDKR

RGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEI

GMKGERRRGKGHDGLYQGLSTATKDTYDALHMOQAL

PPRGSGATNFSLLKQAGDVEENPGPMILNKALMLGAL

ALTTVMSPCGGEDIVADHVASYGVNLYQSYGPSGQYT

HEFDGDEQFYVDLGRKETVWCLPVLRQFRFDPQFALT

NIAVLKHNLNSLIKRSNSTAATNEVPEVTVFSKSPVTLG

QPNILICLVDNIFPPVVNITWLSNGHSVTEGVSETSFLS

KSDHSEFKISYLTLLPSAEESYDCKVEHWGLDKPLLKH

WEPEIPAPMSELTETVVCEWVLVVVGGVLACYSLLVT

VAFHFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQP

YAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNEL

NLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYN

ELOKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTA

TKDTYDALHMQALPPR* 2 Signal AA 32 MSWKKALRIPGGLRAATVTLMLSMLSTPVAEG peptide HLA-

DQB1*02:01 5' Flanking AA 3 QLQ ee 3 HLA-DQ2- AA PFPOPELPY

Ce CP 4 HLA-DQ2- AA PQPELPYPQ me CP

Complete AA PFPQPELPYPQ alpha gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2) 3'Flanking PQL mn

6 AA GSGSGSLGSGSGSGSGS 7 HLA- AA 198 RDSPEDFVYOQOFKGMCYFTNGTERVRLVSRSIYNREEIV

DQB1*02:01 RFDSDVGEFRAVTLLGLPAAEYWNSOKDILERKRAAV

DRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHN

LLVCSVTDFYPAQIKVRWFRNDQEETAGVVSTPLIRNG

DWTFQILVMLEMTPQRGDVYTCHVEHPSLOSPITVE

WRAQSESAQSK

FWVLVVVGGVLACYSLLVTVAFIIFW

CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRD stimulatory FAAYRS domain

CD3 zeta AA 112 RVKESRSADAPAYQQGONOQLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELOQKDKMAEAY signaling SEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ domain ALPPR 11 viral self- AA 19 ATNFSLLKQAGDVEENPGP cleaving 2A polypeptide 12 Signal AA 23 MILNKALMLGALALTTVMSPCGG peptide HLA-

DQA1*05:01 13 HLA- AA 197 EDIVADHVASYGVNLYQSYGPSGQYTHEFDGDEQFYV

DQA1*05:01 DLGRKETVWCLPVLROFRFDPOFALTNIAVLKHNLNSL

IKRSNSTAATNEVPEVTVFSKSPVTLGOPNILICLVDNIF

PPVVNITWLSNGHSVTEGVSETSFLSKSDHSFFKISYLT

LLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSELT

ETVVC

CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRD stimulatory FAAYRS domain 10 CD3 zeta AA 112 RVKFSRSADAPAYQQGONOLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELOKDKMAEAY signaling SEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ fee rT 14 CHAR _HLA- NT WT 2601 ATGTCTTGGAAAAAGGCTTTGCGGATCCCCGGAGG

DQ2.5-glia- CCTTCGGGCAGCAACTGTGACCTTGATGCTGTCGAT ala-glia-02 GCTGAGCACCCCAGTGGCTGAGGGCCAACTTCAAC molecule (a- CTTTTCCTCAACCTGAACTTCCTTATCCTCAACCTCAA gliadin) CTTGGCTCTGGATCTGGGTCCCTGGGATCTGGCTCT

GGATCTGGCTCTGGATCTAGAGACTCTCCCGAGGAT

TTCGTGTACCAGTTTAAGGGCATGTGCTACTTCACC

AACGGGACAGAGCGCGTGCGTCTTGTGAGCAGAAG

CATCTATAACCGAGAAGAGATCGTGCGCTTCGACAG

CGACGTGGGGGAGTTCCGGGCGGTGACGCTGCTG

GGGCTGCCTGCCGCCGAGTACTGGAACAGCCAGAA

GGACATCCTGGAGAGGAAACGGGCGGCGGTGGAC

AGGGTGTGCAGACACAACTACCAGTTGGAGCTCCG

CACGACCTTGCAGCGGCGAGTGGAGCCCACAGTGA

CCATCTCCCCATCCAGGACAGAGGCCCTCAACCACC

ACAACCTGCTGGTCTGCTCGGTGACAGATTTCTATC

CAGCCCAGATCAAAGTCCGGTGGTTTCGGAATGACC

AGGAGGAGACAGCTGGCGTTGTGTCCACCCCCCTTA

TTAGGAATGGTGACTGGACCTTCCAGATCCTGGTGA

TGCTGGAAATGACTCCCCAGCGTGGAGACGTCTACA

CCTGCCACGTGGAGCACCCCAGCCTCCAGAGCCCCA

TCACCGTGGAGTGGCGGGCTCAATCTGAATCTGCCC

AGAGCAAGTTTTGGGTGCTGGTGGTGGTTGGGGGA

GTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCC

TTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAG

GCTCCTGCACAGTGACTACATGAACATGACTCCCCG

CCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTA

TGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAG

AGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGT

ACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTC

AATCTAGGACGAAGAGAGGAGTACGATGTTTTGGA

CAAGAGACGTGGCCGGGACCCTGAGATGGGGGGA

AAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTA

CAATGAACTGCAGAAAGATAAGATGGCGGAGGCCT

ACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAG

GGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCA

GTACAGCCACCAAGGACACCTACGACGCCCTTCACA

TGCAGGCCCTGCCCCCTCGCGGCAGCGGCGCCACC

AACTTCAGCCTGCTGAAGCAGGCCGGCGACGTGGA

GGAAAACCCTGGGCCCATGATCCTAAACAAAGCTCT

GATGCTGGGGGCCCTTGCECTGACCACCGTGATGA

GCCCCTGTGGAGGTGAAGACATTGTGGCTGACCAC

GTCGCCTCTTATGGTGTAAACTTGTACCAGTCTTACG

GTCCCTCTGGCCAGTACACCCATGAATTTGATGGAG

ATGAGCAGTTCTACGTGGACCTGGGGAGGAAGGAG

ACTGTCTGGTGTTTGCCTGTTCTCAGACAATTTAGAT

TTGACCCGCAATTTGCACTGACAAACATCGCTGTCCT

AAAACATAACTTGAACAGTCTGATTAAACGCTCCAA

CTCTACCGCTGCTACCAATGAGGTTCCTGAGGTCAC

AGTGTTTTCCAAGTCTCCCGTGACACTGGGTCAGCC

CAACATCCTCATCTGTCTTGTGGACAACATCTTTCCT

CCTGTGGTCAACATCACATGGCTGAGCAATGGGCAC

TCAGTCACAGAAGGTGTTTCTGAGACCAGCTTCCTC

TCCAAGAGTGATCATTCCTTCTTCAAGATCAGTTACC

TCACCCTCCTCCCTTCTGCTGAGGAGAGTTATGACT

GCAAGGTGGAGCACTGGGGCCTGGACAAGCCTCTT

CTGAAACACTGGGAGCCTGAGATTCCAGCCCCTATG

TCAGAGCTCACAGAGACTGTGGTCTGCTTTTGGGTG

CTGGTGGTGGTTGGGGGAGTCCTGGCTTGCTATAG

CTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTG

AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTA

CATGAACATGACTCCCCGCCGCCCCGGGCCCACCCG

CAAGCATTACCAGCCCTATGCCCCACCACGCGACTT

CGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGA

GCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAAC

CAGCTCTATAACGAGCTCAATCTAGGACGAAGAGA

GGAGTACGATGTTTTGGACAAGAGACGTGGCCGGG

ACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAA

CCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAG

ATAAGATGGCGGAGGCCTACAGTGAGATTGGGATG

AAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATG

GCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACA

CCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTC

GCTGA

Signal NT WT 96 ATGTCTTGGAAAAAGGCTTTGCGGATCCCCGGAGG peptide HLA- CCTTCGGGCAGCAACTGTGACCTTGATGCTGTCGAT

DQB1*02:01 GCTGAGCACCCCAGTGGCTGAGGGC 5' Flanking NT WT CAACTTCAA ee PT 16 HLA-DQ2- NT WT 26 CCTTTTCCTCAACCTGAACTTCCTTA

Fe 17 HLA-DQ2- NT WT 27 CCTCAACCTGAACTTCCTTATCCTCAA ie 18 Complete NT WT 33 CCTTTTCCTCAACCTGAACTTCCTTATCCTCAA gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2) 3'Flanking NT WT CCTCAACTT ee TTT 19 Linker NTWT 51 GGCTCTGGATCTGGGTCCCTGGGATCTGGCTCTGGA ee

HLA- NT WT 594 AGAGACTCTCCCGAGGATTTCGTGTACCAGTTTAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGGACAGAGCGCGT

GCGTCTTGTGAGCAGAAGCATCTATAACCGAGAAG

AGATCGTGCGCTTCGACAGCGACGTGGGGGAGTTC

CGGGCGGTGACGCTGCTGGGGCTGCCTGCCGCCGA

GTACTGGAACAGCCAGAAGGACATCCTGGAGAGGA

AACGGGCGGCGGTGGACAGGGTGTGCAGACACAA

CTACCAGTTGGAGCTCCGCACGACCTTGCAGCGGCG

AGTGGAGCCCACAGTGACCATCTCCCCATCCAGGAC

AGAGGCCCTCAACCACCACAACCTGCTGGTCTGCTC

GGTGACAGATTTCTATCCAGCCCAGATCAAAGTCCG

GTGGTTTCGGAATGACCAGGAGGAGACAGCTGGCG

TTGTGTCCACCCCCCTTATTAGGAATGGTGACTGGA

CCTTCCAGATCCTGGTGATGCTGGAAATGACTCCCC

AGCGTGGAGACGTCTACACCTGCCACGTGGAGCAC

CCCAGCCTCCAGAGCCCCATCACCGTGGAGTGGCG

GGCTCAATCTGAATCTGCCCAGAGCAAG

21 CD28 TM NT WT 78 TITTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATT

TICTGG

22 CD28 co- NT WT 126 GTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGA stimulatory CTACATGAACATGACTCCCCGCCGCCCCGGGCCCAC domain CCGCAAGCATTACCAGCCCTATGCCCCACCACGCGA

CTTCGCAGCCTATCGCTCC

23 CD3 zeta NT WT 336 AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGC primary GTACCAGCAGGGCCAGAACCAGCTCTATAACGAGC signaling TCAATCTAGGACGAAGAGAGGAGTACGATGITTIG domain GACAAGAGACGTGGCCGGGACCCTGAGATGGGGG

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAGG

CCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGG

AGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT

CAGTACAGCCACCAAGGACACCTACGACGCCCTTCA

CATGCAGGCCCTGCCCCCTCGC

24 viral self- NT WT 57 GCCACCAACTTCAGCCTGCTGAAGCAGGCCGGCGA cleaving 2A CGTGGAGGAAAACCCTGGGCCC polypeptide

Signal NT WT ATGATCCTAAACAAAGCTCTGATGCTGGGGGCCCTT peptide HLA- GCCCTGACCACCGTGATGAGCECCTGTGGAGGT

DQA1*05:01 26 HLA- NT WT 591 GAAGACATTGTGGCTGACCACGTCGCCTCTTATGGT

DQA1*05:01 GTAAACTTGTACCAGTCTTACGGTCCCTCTGGCCAG

TACACCCATGAATTTGATGGAGATGAGCAGTTCTAC

GTGGACCTGGGGAGGAAGGAGACTGTCTGGTGTTT

GCCTGTTCTCAGACAATTTAGATTTGACCCGCAATTT

GCACTGACAAACATCGCTGTCCTAAAACATAACTTG

AACAGTCTGATTAAACGCTCCAACTCTACCGCTGCT

ACCAATGAGGTTCCTGAGGTCACAGTGTTTTCCAAG

TCTCCCGTGACACTGGGTCAGCCCAACATCCTCATCT

GTCTTGTGGACAACATCTTTCCTCCTGTGGTCAACAT

CACATGGCTGAGCAATGGGCACTCAGTCACAGAAG

GTGTTTCTGAGACCAGCTTCCTCTCCAAGAGTGATC

ATTCCTTCTTCAAGATCAGTTACCTCACCCTCCTCCCT

TCTGCTGAGGAGAGTTATGACTGCAAGGTGGAGCA

CTGGGGCCTGGACAAGCCTCTTCTGAAACACTGGG

AGCCTGAGATTCCAGCCCCTATGTCAGAGCTCACAG

AGACTGTGGTCTGC

21 CD28 TM NT WT 78 TTTTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATT

TTCTGG

CTTCGCAGCCTATCGCTCC

23 CD3 zeta NT WT 336 AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCECGC primary GTACCAGCAGGGCCAGAACCAGCTCTATAACGAGC signaling TCAATCTAGGACGAAGAGAGGAGTACGATGTTTTG domain GACAAGAGACGTGGCCGGGACCCTGAGATGGGGG

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAGG

CCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGG

AGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCT

CAGTACAGCCACCAAGGACACCTACGACGCCCTTCA

CATGCAGGCCCTGCCCCCTCGC he TGA 27 CHAR_HLA- | NT OPT 2601 | ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG

DQ2.5-glia- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT ala-glia-a2 GCTGAGCACCCCGGTGGCGGAGGGTCAGCTGCAAC molecule {o- CGTTTCCGCAGCCGGAACTGCCGTACCCGCAGCCGC gliadin) AACTGGGTAGCGGTAGCGGTAGCCTGGGCAGCGGC

AGCGGTAGCGGTAGCGGCAGCCGTGACAGCCCGG

AGGATTTCGTTTACCAATTCAAGGGCATGTGCTACT

TCACCAACGGCACCGAACGTGTGCGTCTGGTTAGCC

GTAGCATCTACAACCGTGAGGAAATTGTGCGTTTCG

ACAGCGATGTTGGCGAGTTTCGTGCGGTGACCCTG

CTGGGTCTGCCGGCGGCGGAGTACTGGAACAGCCA

GAAGGACATCCTGGAACGTAAACGTGCGGCGGTGG

ATCGTGTTTGCCGTCACAACTATCAGCTGGAGCTGC

GTACCACCCTGCAACGTCGTGTGGAACCGACCGTTA

CCATCAGCCCGAGCCGTACCGAAGCGCTGAACCACC

ACAACCTGCTGGTGTGCAGCGTTACCGACTTCTACC

CGGCGCAGATTAAAGTTCGTTGGTTTCGTAACGATC

AAGAGGAAACCGCGGGTGTGGTTAGCACCCCGCTG

ATCCGTAACGGCGACTGGACCTTCCAGATTCTGGTT

ATGCTGGAGATGACCCCGCAACGTGGTGATGTGTA

CACCTGCCACGTTGAACACCCGAGCCTGCAGAGCCC

GATTACCGTGGAGTGGCGTGCGCAGAGCGAAAGCG

CGCAAAGCAAGTTTTGGGTTCTGGTGGTTGTGGGT

GGCGTGCTGGCGTGCTACAGCCTGCTGGTGACCGT

TGCGTTCATCATCTTCTGGGTGCGTAGCAAACGTAG

CCGTCTGCTGCACAGCGACTATATGAACATGACCCC

GCGTCGTCCGGGTCCGACCCGTAAGCACTACCAACC

GTATGCGCCGCCGCGTGACTTTGCGGCGTACCGTA

GCCGTGTTAAATTTAGCCGTAGCGCGGATGCGCCG

GCGTACCAGCAGGGTCAGAACCAACTGTATAACGA

GCTGAACCTGGGCCGTCGTGAGGAATATGACGTGC

TGGATAAGCGTCGTGGTCGTGATCCGGAAATGGGT

GGCAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCT

GTACAACGAACTGCAAAAGGACAAAATGGCGGAG

GCGTATAGCGAAATTGGTATGAAGGGCGAGCGTCG

TCGTGGTAAAGGCCACGATGGTCTGTACCAGGGCC

TGAGCACCGCGACCAAAGACACCTATGATGCGCTG

CACATGCAAGCGCTGCCGCCGCGTGGTAGCGGTGC

GACCAACTTCAGCCTGCTGAAGCAGGCGGGTGACG

TTGAGGAAAACCCGGGCCCGATGATCCTGAACAAA

GCGCTGATGCTGGGTGCGCTGGCGCTGACCACCGT

TATGAGCCCGTGCGGTGGCGAGGACATTGTGGCGG

ATCACGTTGCGAGCTACGGCGTGAACCTGTACCAGA

GCTATGGTCCGAGCGGCCAATACACCCACGAGTTCG

ACGGTGATGAACAATTITATGTTGACCTGGGCCGTA

AGGAAACCGTGTGGTGCCTGCCGGTTCTGCGTCAG

TTCCGTTTTGATCCGCAATTCGCGCTGACCAACATCG

CGGTGCTGAAGCACAACCTGAACAGCCTGATTAAAC

GTAGCAACAGCACCGCGGCGACCAACGAGGTTCCG

GAAGTGACCGTTTTCAGCAAAAGCCCGGTGACCCTG

GGTCAGCCGAACATCCTGATTTGCCTGGTTGACAAC

ATCTTTCCGCCGGTTGTGAACATTACCTGGCTGAGC

AACGGTCACAGCGTGACCGAGGGCGTTAGCGAAAC

CAGCTTCCTGAGCAAGAGCGATCACAGCTTCTTTAA

AATCAGCTATCTGACCCTGCTGCCGAGCGCGGAGG

AAAGCTATGACTGCAAGGTGGAGCACTGGGGTCTG

GATAAGCCGCTGCTGAAACACTGGGAGCCGGAAAT

TCCGGCGCCGATGAGCGAGCTGACCGAAACCGTTG

TGTGCTTTTGGGTTCTGGTTGTGGTTGGTGGCGTGT

TAGCTTGCTATAGCCTGCTGGTTACCGTGGCGTTTA

TTATCTTCTGGGTTCGCAGCAAGCGTAGCCGTCTGC

TGCATAGCGATTACATGAATATGACCCCGCGTCGTC

CTGGCCCGACCCGCAAACATTATCAACCGTACGCGC

CGCCGCGTGACTTTGCAGCGTATCGTAGCCGTGTTA

AGTTTAGCCGTAGCGCGGACGCGCCGGCGTATCAA

CAGGGCCAAAATCAGCTGTACAATGAACTGAATCTG

GGTCGTCGTGAAGAGTACGATGTTCTGGACAAACG

TCGTGGTCGTGACECGGAGATGGGTGGCAAACCGC

GTCGTAAGAACCCGCAGGAAGGTTTATATAATGAG

CTGCAGAAAGATAAGATGGCGGAAGCGTATAGCGA

AATCGGTATGAAGGGCGAACGTCGTCGTGGCAAGG

GTCATGACGGCCTGTATCAAGGTCTGAGCACCGCG

ACCAAGGATACCTACGACGCGCTGCATATGCAGGC

GCTGCCGCCGCGTTAA

28 Signal NT OPT ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG peptide HLA- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT

DQB1*02:01 GCTGAGCACCCCGGTGGCGGAGGGT 5' Flanking NT OPT CAGCTGCAA ee TT 29 HLA-DQ2- NT OPT 27 CCGTTTCCGCAGCCGGAACTGCCGTAC rn

HLA-DQ2- NT OPT 27 CCGCAGCCGGAACTGCCGTACCCGCAG ee [TTT 31 Complete NT OPT 33 CCGTTTCCGCAGCCGGAACTGCCGTACCCGCAG gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2) 3'Flanking NT OPT 9 CCGCAACTG ee TTT 32 Linker NT OPT 51 GGTAGCGGTAGCGGTAGCCTGGGCAGCGGCAGCG

TOP ee 33 HLA- NT OPT 594 CGTGACAGCCCGGAGGATTTCGTTTACCAATTCAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGCACCGAACGTGTG

CGTCTGGTTAGCCGTAGCATCTACAACCGTGAGGAA

ATTGTGCGTTTCGACAGCGATGTTGGCGAGTTTCGT

GCGGTGACCCTGCTGGGTCTGCCGGCGGCGGAGTA

CTGGAACAGCCAGAAGGACATCCTGGAACGTAAAC

GTGCGGCGGTGGATCGTGTTTGCCGTCACAACTATC

AGCTGGAGCTGCGTACCACCCTGCAACGTCGTGTG

GAACCGACCGTTACCATCAGCCCGAGCCGTACCGAA

GCGCTGAACCACCACAACCTGCTGGTGTGCAGCGTT

ACCGACTTCTACCCGGCGCAGATTAAAGTTCGTTGG

TTTCGTAACGATCAAGAGGAAACCGCGGGTGTGGT

TAGCACCCCGCTGATCCGTAACGGCGACTGGACCTT

CCAGATTCTGGTTATGCTGGAGATGACCCCGCAACG

TGGTGATGTGTACACCTGCCACGTTGAACACCCGAG

CCTGCAGAGCCCGATTACCGTGGAGTGGCGTGCGC

AGAGCGAAAGCGCGCAAAGCAAG

34 CD28 TM NT OPT 78 TTTTGGGTTCTGGTGGTTGTGGGTGGCGTGCTGGC

GTGCTACAGCCTGCTGGTGACCGTTGCGTTCATCAT

CTTCTGG

CD28 co- NT OPT 126 GTGCGTAGCAAACGTAGCCGTCTGCTGCACAGCGA stimulatory CTATATGAACATGACCCCGCGTCGTCCGGGTCCGAC domain CCGTAAGCACTACCAACCGTATGCGCCGCCGCGTGA

CTTTGCGGCGTACCGTAGC

CD3 zeta NT OPT 336 CGTGTTAAATTTAGCCGTAGCGCGGATGCGCCGGC primary GTACCAGCAGGGTCAGAACCAACTGTATAACGAGC signaling TGAACCTGGGCCGTCGTGAGGAATATGACGTGCTG domain GATAAGCGTCGTGGTCGTGATCCGGAAATGGGTGG

CAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCTGT

ACAACGAACTGCAAAAGGACAAAATGGCGGAGGC

GTATAGCGAAATTGGTATGAAGGGCGAGCGTCGTC

GTGGTAAAGGCCACGATGGTCTGTACCAGGGCCTG

AGCACCGCGACCAAAGACACCTATGATGCGCTGCA

CATGCAAGCGCTGCCGCCGCGT he NT OPT 9 GGTAGCGGT 37 viral self- NT OPT 57 | GCGACCAACTTCAGCCTGCTGAAGCAGGCGGGTGA cleaving 2A CGTTGAGGAAAACCCGGGCCCG polypeptide

38 Signal NT OPT ATGATCCTGAACAAAGCGCTGATGCTGGGTGCGCT peptide HLA- GGCGCTGACCACCGTTATGAGCCCGTGCGGTGGC

DQA1*05:01 42 HLA- NT OPT 591 GAGGACATTGTGGCGGATCACGTTGCGAGCTACGG

DQA1*05:01 CGTGAACCTGTACCAGAGCTATGGTCCGAGCGGCC

AATACACCCACGAGTTCGACGGTGATGAACAATTTT

ATGTTGACCTGGGCCGTAAGGAAACCGTGTGGTGC

CTGCCGGTTCTGCGTCAGTTCCGTTTTGATCCGCAAT

TCGCGCTGACCAACATCGCGGTGCTGAAGCATAACC

TGAACAGCCTGATTAAACGTAGCAACAGCACCGCG

GCGACCAACGAGGTTCCGGAAGTGACCGTTTTCAG

CAAAAGCCCGGTGACCCTGGGTCAGCCGAACATCCT

GATTTGCCTGGTTGACAACATCTTTCCGCCGGTTGT

GAACATTACCTGGCTGAGCAACGGTCACAGCGTGA

CCGAGGGCGTTAGCGAAACCAGCTTCCTGAGCAAG

AGCGATCACAGCTTCTTTAAAATCAGCTATCTGACCC

TGCTGCCGAGCGCGGAGGAAAGCTATGACTGCAAG

GTGGAGCACTGGGGTCTGGATAAGCCGCTGCTGAA

ACACTGGGAGCCGGAAATTCCGGCGCCGATGAGCG

AGCTGACCGAAACCGTTGTGTGC

39 CD28 TM NT OPT 78 TTTTGGGTTCTGGTTGTGGTTGGTGGCGTGTTAGCT

TGCTATAGCCTGCTGGTTACCGTGGCGTTTATTATCT

TCTGG

CD28 co- NT OPT 126 GTTCGCAGCAAGCGTAGCCGTCTGCTGCATAGCGAT stimulatory TACATGAATATGACCCCGCGTCGTCCTGGCCCGACC domain CGCAAACATTATCAACCGTACGCGCCGCCGCGTGAC

TTTGCAGCGTATCGTAGC

41 CD3 zeta NT OPT 336 CGTGTTAAGTTTAGCCGTAGCGCGGACGCGCCGGC primary GTATCAACAGGGCCAAAATCAGCTGTACAATGAACT signaling GAATCTGGGTCGTCGTGAAGAGTACGATGTTCTGG domain ACAAACGTCGTGGTCGTGACCCGGAGATGGGTGGC

AAACCGCGTCGTAAGAACCCGCAGGAAGGTTTATA

TAATGAGCTGCAGAAAGATAAGATGGCGGAAGCGT

ATAGCGAAATCGGTATGAAGGGCGAACGTCGTCGT

GGCAAGGGTCATGACGGCCTGTATCAAGGTCTGAG

CACCGCGACCAAGGATACCTACGACGCGCTGCATAT

GCAGGCGCTGCCGCCGCGT

Table 2: CHAR HLA-DQ2.5-glia-Q1-glia-Q2 molecule

CHAR Amino Acid (AA), Length | Sequence

NO POLYPEPTIDE Nucleotide (NT) or

Nucleotide Codon

Optimised (NT OPT} 43 CHAR _HLA- AA 866 MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGPGOP

DQ2.5-glia- FPOPEQPFPWOPQGSGSGSLGSGSGSGSGSRDSPED

Q1-glia-02 FVYQFKGMCYFTNGTERVRLVSRSIYNREEIVREDSDV molecule (Q- GEFRAVTLLGLPAAEYWNSQKDILERKRAAVDRVCRH gliadin) NYOQLELRTTLORRVEPTVTISPSRTEALNHHNLLVCSVT

DFYPAQIKVRWERNDOQEETAGVVSTPLIRNGDWTFQI

LVMLEMTPOQRGDVYTCHVEHPSLQSPITVEWRAGSE

SAQSKFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSR

LLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSR

VKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDK

RRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYS

EIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ

ALPPRGSGATNFSLLKQAGDVEENPGPMILNKALML

GALALTTVMSPCGGEDIVADHVASYGVYNLYQSYGPS

GQYTHEFDGDEQFYVDLGRKETVWCLPVLRQFRFDP

QFALTNIAVLKHNLNSLIKRSNSTAATNEVPEVTVFSKS

PVTLGOPNILICLVDNIFPPVVNITWLSNGHSVTEGVS

ETSFLSKSDHSFFKISYLTLLPSAEESYDCKVEHWGLDK

PLLKHWEPEIPAPMSELTETVVCFWVLVVVGGVLACY

SLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRK

HYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGONQL

YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQE

GLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQG

LSTATKDTYDALHMOQALPPR

2 Signal AA 32 MSWKKALRIPGGLRAATVTLMLSMLSTPVAEG peptide HLA-

DQB1*02:01 5' Flanking AA 3 PQQ sequence BN 44 HLA-DQ2- AA PFPQPEQPF

Ce 45 HLA-DQ2- AA PQPEQPFPW ie 46 Complete AA 11 PFPQPEQPFPW gliadin omega peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) 3'Flanking AA 3 en 7 HLA- AA 198 RDSPEDFVYQOFKGMCYFTNGTERVRLVSRSIYNREEIV

DQB1*02:01 RFDSDVGEFRAVTLLGLPAAEYWNSOQKDILERKRAAV

DRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHN

LLVCSVTDFYPAQIKVRWFRNDQEETAGVVSTPLIRN

GDWTFQILVMLEMTPQRGDVYTCHVEHPSLQSPITV

EWRAQSESAQSK

CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYOQPYAPPRD stimulatory FAAYRS domain

CD3 zeta AA 112 RVKFSRSADAPAYQQGONOLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAY signaling SEIGMKGERRRGKGHDGLYOGLSTATKDTYDALHMQ domain ALPPR

11 viral self- AA 19 ATNFSLLKQAGDVEENPGP cleaving 2A polypeptide 12 Signal AA 23 MILNKALMLGALALTTVMSPCGG peptide HLA-

DQA1*05:01 13 HLA- AA 197 EDIVADHVASYGVNLYQSYGPSGQYTHEFDGDEQFYV

DQA1*05:01 DLGRKETVWCLPVLRQFRFDPQFALTNIAVLKHNLNS

LIKRSNSTAATNEVPEVTVFSKSPVTLGQPNILICLVDNI

FPPVVNITWLSNGHSVTEGVSETSFLSKSDHSFFKISYL

TLLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSEL

TETVVC

Te | a | ae 9 CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRD stimulatory FAAYRS domain

CD3 zeta AA 112 | RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELOKDKMAEAY signaling SEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ domain ALPPR 47 CHAR _HLA- NT WT 2601 | ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG

DQ2.5-glia- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT

Q1-glia-Q2 GCTGAGCACCCCGGTGGCGGAGGGTCCACAACAAC molecule (Q- CTTTTCCACAGCCGGAACAACCATTTCCCTGGCAAC gliadin) CACAAGGTAGCGGTAGCGGTAGCCTGGGCAGCGG

CAGCGGTAGCGGTAGCGGCAGCCGTGACAGCCCG

GAGGATTTCGTTTACCAATTCAAGGGCATGTGCTAC

TTCACCAACGGCACCGAACGTGTGCGTCTGGTTAGC

CGTAGCATCTACAACCGTGAGGAAATTGTGCGTTTC

GACAGCGATGTTGGCGAGTTTCGTGCGGTGACCCT

GCTGGGTCTGCCGGCGGCGGAGTACTGGAACAGCC

AGAAGGACATCCTGGAACGTAAACGTGCGGCGGTG

GATCGTGTTTGCCGTCACAACTATCAGCTGGAGCTG

CGTACCACCCTGCAACGTCGTGTGGAACCGACCGTT

ACCATCAGCCCGAGCCGTACCGAAGCGCTGAACCA

CCACAACCTGCTGGTGTGCAGCGTTACCGACTTCTA

CCCGGCGCAGATTAAAGTTCGTTGGTTTCGTAACGA

TCAAGAGGAAACCGCGGGTGTGGTTAGCACCCCGC

TGATCCGTAACGGCGACTGGACCTTCCAGATTCTGG

TTATGCTGGAGATGACCCCGCAACGTGGTGATGTGT

ACACCTGCCACGTTGAACACCCGAGCCTGCAGAGCC

CGATTACCGTGGAGTGGCGTGCGCAGAGCGAAAGC

GCGCAAAGCAAGTTTTGGGTTCTGGTGGTTGTGGG

TGGCGTGCTGGCGTGCTACAGCCTGCTGGTGACCG

TTGCGTTCATCATCTTCTGGGTGCGTAGCAAACGTA

GCCGTCTGCTGCACAGCGACTATATGAACATGACCC

CGCGTCGTCCGGGTCCGACCCGTAAGCACTACCAAC

CGTATGCGCCGCCGCGTGACTTTGCGGCGTACCGTA

GCCGTGTTAAATTTAGCCGTAGCGCGGATGCGCCG

GCGTACCAGCAGGGTCAGAACCAACTGTATAACGA

GCTGAACCTGGGCCGTCGTGAGGAATATGACGTGC

TGGATAAGCGTCGTGGTCGTGATCCGGAAATGGGT

GGCAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCT

GTACAACGAACTGCAAAAGGACAAAATGGCGGAG

GCGTATAGCGAAATTGGTATGAAGGGCGAGCGTCG

TCGTGGTAAAGGCCACGATGGTCTGTACCAGGGCC

TGAGCACCGCGACCAAAGACACCTATGATGCGCTG

CACATGCAAGCGCTGCCGCCGCGTGGTAGCGGTGC

GACCAACTTCAGCCTGCTGAAGCAGGCGGGTGACG

TTGAGGAAAACCCGGGCCCGATGATCCTGAACAAA

GCGCTGATGCTGGGTGCGCTGGCGCTGACCACCGT

TATGAGCCCGTGCGGTGGCGAGGACATTGTGGCGG

ATCACGTTGCGAGCTACGGCGTGAACCTGTACCAG

AGCTATGGTCCGAGCGGCCAATACACCCACGAGTTC

GACGGTGATGAACAATTTTATGTTGACCTGGGCCGT

AAGGAAACCGTGTGGTGCCTGCCGGTTCTGCGTCA

GTTCCGTTTTGATCCGCAATTCGCGCTGACCAACATC

GCGGTGCTGAAGCACAACCTGAACAGCCTGATTAA

ACGTAGCAACAGCACCGCGGCGACCAACGAGGTTC

CGGAAGTGACCGTTTTCAGCAAAAGCCCGGTGACC

CTGGGTCAGCCGAACATCCTGATTTGCCTGGTTGAC

AACATCTTTCCGCEGGTTGTGAACATTACCTGGCTG

AGCAACGGTCACAGCGTGACCGAGGGCGTTAGCGA

AACCAGCTTCCTGAGCAAGAGCGATCACAGCTTCTT

TAAAATCAGCTATCTGACCCTGCTGCCGAGCGCGGA

GGAAAGCTATGACTGCAAGGTGGAGCACTGGGGTC

TGGATAAGCCGCTGCTGAAACACTGGGAGCCGGAA

ATTCCGGCGCCGATGAGCGAGCTGACCGAAACCGT

TGTGTGCTTTTGGGTTCTGGTTGTGGTTGGTGGCGT

GTTAGCTTGCTATAGCCTGCTGGTTACCGTGGCGTT

TATTATCTTCTGGGTTCGCAGCAAGCGTAGCCGTCT

GCTGCATAGCGATTACATGAATATGACCCCGCGTCG

TCCTGGCCCGACCCGCAAACATTATCAACCGTACGC

GCCGCCGCGTGACTTTGCAGCGTATCGTAGCCGTGT

TAAGTTTAGCCGTAGCGCGGACGCGCCGGCGTATC

AACAGGGCCAAAATCAGCTGTACAATGAACTGAAT

CTGGGTCGTCGTGAAGAGTACGATGTTCTGGACAA

ACGTCGTGGTCGTGACCCGGAGATGGGTGGCAAAC

CGCGTCGTAAGAACCCGCAGGAAGGTTTATATAAT

GAGCTGCAGAAAGATAAGATGGCGGAAGCGTATA

GCGAAATCGGTATGAAGGGCGAACGTCGTCGTGGC

AAGGGTCATGACGGCCTGTATCAAGGTCTGAGCAC

CGCGACCAAGGATACCTACGACGCGCTGCATATGC

AGGCGCTGCCGCCGCGTTAA

Signal NT WT ATGTCTTGGAAAAAGGCTTTGCGGATCCCCGGAGG peptide HLA- CCTTCGGGCAGCAACTGTGACCTTGATGCTGTCGAT

DQB1*02:01 GCTGAGCACCCCAGTGGCTGAGGGC 5' Flanking NT WT CCACAACAA sequence BN

48 HLA-DQ2- NT WT CCTTTTCCACAGCCGGAACAACCATTT ln | TTT 49 HLA-DQ2- CCACAGCCGGAACAACCATTTCCCTGG an em 50 Complete NT WT 33 CCTTTTCCACAGCCGGAACAACCATTTCCCTGG gliadin peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) 3'Flanking NT WT CAACCACAA le | TT 19 Linker NT WT 51 GGCTCTGGATCTGGGTECCTGGGATCTGGCTCTGG

HLA- NT WT 594 | AGAGACTCTCCCGAGGATTTCGTGTACCAGTTTAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGGACAGAGCGCGT

GCGTCTTGTGAGCAGAAGCATCTATAACCGAGAAG

AGATCGTGCGCTTCGACAGCGACGTGGGGGAGTTIC

CGGGCGGTGACGCTGCTGGGGCTGCCTGCCGCCGA

GTACTGGAACAGCCAGAAGGACATCCTGGAGAGGA

AACGGGCGGCGGTGGACAGGGTGTGCAGACACAA

CTACCAGTTGGAGCTCCGCACGACCTTGCAGCGGC

GAGTGGAGCCCACAGTGACCATCTCCCCATCCAGG

ACAGAGGCCCTCAACCACCACAACCTGCTGGTCTGC

TCGGTGACAGATTTCTATCCAGCCCAGATCAAAGTC

CGGTGGTTTCGGAATGACCAGGAGGAGACAGCTGG

CGTTGTGTCCACCCCCCTTATTAGGAATGGTGACTG

GACCTTCCAGATCCTGGTGATGCTGGAAATGACTCC

CCAGCGTGGAGACGTCTACACCTGCCACGTGGAGC

ACCCCAGCCTCCAGAGCCCCATCACCGTGGAGTGGC

GGGCTCAATCTGAATCTGCCCAGAGCAAG

21 CD28 TM NT WT 78 TTTTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATT

TTCTGG

22 CD28 co- NT WT 126 | GTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTG stimulatory ACTACATGAACATGACTCCCCGCCGCCCCGGGCCCA domain CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCG

ACTTCGCAGCCTATCGCTCC

23 CD3 zeta NT WT 336 | AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGC primary GTACCAGCAGGGCCAGAACCAGCTCTATAACGAGC signaling TCAATCTAGGACGAAGAGAGGAGTACGATGTTTTG domain GACAAGAGACGTGGCCGGGACCCTGAGATGGGGG

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAG

GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCG

GAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTC

TCAGTACAGCCACCAAGGACACCTACGACGCCCTTC

ACATGCAGGCCCTGCCCCCTCGC

24 viral self- NT WT 57 GCCACCAACTTCAGCCTGCTGAAGCAGGCCGGCGA cleaving 2A CGTGGAGGAAAACCCTGGGLCC polypeptide

Signal NT WT ATGATCCTAAACAAAGCTCTGATGCTGGGGGCCCTT peptide HLA- GCCCTGACCACCGTGATGAGCCCCTGTGGAGGT

DQA1*05:01 26 HLA- GAAGACATTGTGGCTGACCACGTCGCCTCTTATGGT

DQA1*05:01 GTAAACTTGTACCAGTCTTACGGTCCCTCTGGCCAG

TACACCCATGAATTTGATGGAGATGAGCAGTTCTAC

GTGGACCTGGGGAGGAAGGAGACTGTCTGGTGTTT

GCCTGTTCTCAGACAATTTAGATTTGACCCGCAATTIT

GCACTGACAAACATCGCTGTCCTAAAACATAACTTG

AACAGTCTGATTAAACGCTCCAACTCTACCGCTGCT

ACCAATGAGGTTCCTGAGGTCACAGTGTTTTCCAAG

TCTCCCGTGACACTGGGTCAGCCCAACATCCTCATCT

GTCTTGTGGACAACATCTTTCCTCCTGTGGTCAACAT

CACATGGCTGAGCAATGGGCACTCAGTCACAGAAG

GTGTTTCTGAGACCAGCTTECTCTCCAAGAGTGATC

ATTCCTTCTTCAAGATCAGTTACCTCACCCTCCTCCCT

TCTGCTGAGGAGAGTTATGACTGCAAGGTGGAGCA

CTGGGGCCTGGACAAGCCTCTTCTGAAACACTGGG

AGCCTGAGATTCCAGCCCCTATGTCAGAGCTCACAG

AGACTGTGGTCTGC

21 CD28 TM NT WT 78 TTTTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATT

TTCTGG

22 CD28 co- NT WT 126 | GTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTG stimulatory ACTACATGAACATGACTCCCCGCCGCCECGGGCCCA domain CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCG

ACTTCGCAGCCTATCGCTCC

23 CD3 zeta NT WT 336 | AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGC primary GTACCAGCAGGGCCAGAACCAGCTCTATAACGAGC signaling TCAATCTAGGACGAAGAGAGGAGTACGATGTITTG domain GACAAGAGACGTGGCCGGGACCCTGAGATGGGGG

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAG

GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCG

GAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTC

TCAGTACAGCCACCAAGGACACCTACGACGCCCTIC

ACATGCAGGCCCTGCCCCCTEGC

51 CHAR _HLA- NT OPT 2601 | ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG

DQ2.5-glia- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT

Q1-glia-Q2 GCTGAGCACCCCGGTGGCGGAGGGTCCACAGCAGC molecule {Q- CCTTCCCTCAGCCAGAGCAGCCCTTTCCTTGGCAGC gliadin) CCCAGGGTAGCGGTAGCGGTAGCCTGGGCAGCGG

CAGCGGTAGCGGTAGCGGCAGCCGTGACAGCCCG

GAGGATTTCGTTTACCAATTCAAGGGCATGTGCTAC

TTCACCAACGGCACCGAACGTGTGCGTCTGGTTAGC

CGTAGCATCTACAACCGTGAGGAAATTGTGCGTTTC

GACAGCGATGTTGGCGAGTTTCGTGCGGTGACCCT

GCTGGGTCTGCCGGCGGCGGAGTACTGGAACAGCC

AGAAGGACATCCTGGAACGTAAACGTGCGGCGGTG

GATCGTGTTTGCCGTCACAACTATCAGCTGGAGCTG

CGTACCACCCTGCAACGTCGTGTGGAACCGACCGTT

ACCATCAGCCCGAGCCGTACCGAAGCGCTGAACCA

CCACAACCTGCTGGTGTGCAGCGTTACCGACTTCTA

CCCGGCGCAGATTAAAGTTCGTTGGTTTCGTAACGA

TCAAGAGGAAACCGCGGGTGTGGTTAGCACCCCGC

TGATCCGTAACGGCGACTGGACCTTCCAGATTCTGG

TTATGCTGGAGATGACCCCGCAACGTGGTGATGTGT

ACACCTGCCACGTTGAACACCCGAGCCTGCAGAGCC

CGATTACCGTGGAGTGGCGTGCGCAGAGCGAAAGC

GCGCAAAGCAAGTTTTGGGTTCTGGTGGTTGTGGG

TGGCGTGCTGGCGTGECTACAGCCTGCTGGTGACCG

TTGCGTTCATCATCTTCTGGGTGCGTAGCAAACGTA

GCCGTCTGCTGCACAGCGACTATATGAACATGACCC

CGCGTCGTCEGGGTCCGACCCGTAAGCACTACCAAC

CGTATGCGCCGCCGCGTGACTTTGCGGCGTACCGTA

GCCGTGTTAAATTTAGCCGTAGCGCGGATGCGCCG

GCGTACCAGCAGGGTCAGAACCAACTGTATAACGA

GCTGAACCTGGGCCGTCGTGAGGAATATGACGTGC

TGGATAAGCGTCGTGGTCGTGATCCGGAAATGGGT

GGCAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCT

GTACAACGAACTGCAAAAGGACAAAATGGCGGAG

GCGTATAGCGAAATTGGTATGAAGGGCGAGCGTCG

TCGTGGTAAAGGCCACGATGGTCTGTACCAGGGCC

TGAGCACCGCGACCAAAGACACCTATGATGCGCTG

CACATGCAAGCGCTGCCGCCGCGTGGTAGCGGTGC

GACCAACTTCAGCCTGCTGAAGCAGGCGGGTGACG

TTGAGGAAAACCCGGGCCCGATGATCCTGAACAAA

GCGCTGATGCTGGGTGCGCTGGCGCTGACCACCGT

TATGAGCCCGTGCGGTGGCGAGGACATTGTGGCGG

ATCACGTTGCGAGCTACGGCGTGAACCTGTACCAG

AGCTATGGTCCGAGCGGCCAATACACCCACGAGTTC

GACGGTGATGAACAATTTTATGTTGACCTGGGCCGT

AAGGAAACCGTGTGGTGCCTGCCGGTTCTGCGTCA

GTTCCGTTTTGATCCGCAATTCGCGCTGACCAACATC

GCGGTGCTGAAGCACAACCTGAACAGCCTGATTAA

ACGTAGCAACAGCACCGCGGCGACCAACGAGGTTC

CGGAAGTGACCGTTTTCAGCAAAAGCCCGGTGACC

CTGGGTCAGCCGAACATCCTGATTTGCCTGGTTGAC

AACATCTTTCCGCCGGTTGTGAACATTACCTGGCTG

AGCAACGGTCACAGCGTGACCGAGGGCGTTAGCGA

AACCAGCTTCCTGAGCAAGAGCGATCACAGCTTCTT

TAAAATCAGCTATCTGACCCTGCTGCCGAGCGCGGA

GGAAAGCTATGACTGCAAGGTGGAGCACTGGGGTC

TGGATAAGCCGCTGCTGAAACACTGGGAGCCGGAA

ATTCCGGCGCCGATGAGCGAGCTGACCGAAACCGT

TGTGTGCTTTTGGGTTCTGGTTGTGGTTGGTGGCGT

GTTAGCTTGCTATAGCCTGCTGGTTACCGTGGCGTT

TATTATCTTCTGGGTTCGCAGCAAGCGTAGCCGTCT

GCTGCATAGCGATTACATGAATATGACCCCGCGTCG

TCCTGGCCCGACCCGCAAACATTATCAACCGTACGC

GCCGCCGCGTGACTTTGCAGCGTATCGTAGCCGTGT

TAAGTTTAGCCGTAGCGCGGACGCGCCGGCGTATC

AACAGGGCCAAAATCAGCTGTACAATGAACTGAAT

CTGGGTCGTCGTGAAGAGTACGATGTTCTGGACAA

ACGTCGTGGTCGTGACCCGGAGATGGGTGGCAAAC

CGCGTCGTAAGAACCCGCAGGAAGGTTTATATAAT

GAGCTGCAGAAAGATAAGATGGCGGAAGCGTATA

GCGAAATCGGTATGAAGGGCGAACGTCGTCGTGGC

AAGGGTCATGACGGCCTGTATCAAGGTCTGAGCAC

CGCGACCAAGGATACCTACGACGCGCTGCATATGC

AGGCGCTGCCGCCGCGTTAA

DQB1*02:01 GCTGAGCACCCCGGTGGCGGAGGGT

5' Flanking NT OPT CCACAGCAG le TT 52 HLA-DQ2- CCCTTCCCTCAGCCAGAGCAGCCCTTT

Cen ee 53 HLA-DQ2- NT OPT 27 CCTCAGCCAGAGCAGCCCTTTCCTTGG glia-Q2 7 54 Complete NT OPT 33 CCCTTCCCTCAGCCAGAGCAGCCCTTTCCTTGG gliadin peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) 3'Flanking NT OPT CAGCCCCAG ee 32 Linker NT OPT 51 GGTAGCGGTAGCGGTAGCCTGGGCAGCGGCAGCG vO GTAGCGGTAGCGGCAGC 33 HLA- NT OPT 594 | CGTGACAGCCCGGAGGATTTCGTTTACCAATTCAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGCACCGAACGTGTG

CGTCTGGTTAGCCGTAGCATCTACAACCGTGAGGAA

ATTGTGCGTTTCGACAGCGATGTTGGCGAGTTTCGT

GCGGTGACCCTGCTGGGTCTGCCGGCGGCGGAGTA

CTGGAACAGCCAGAAGGACATCCTGGAACGTAAAC

GTGCGGCGGTGGATCGTGTTTGCCGTCACAACTATC

AGCTGGAGCTGCGTACCACCCTGCAACGTCGTGTG

GAACCGACCGTTACCATCAGCCCGAGCCGTACCGA

AGCGCTGAACCACCACAACCTGCTGGTGTGCAGCG

TTACCGACTTCTACCCGGCGCAGATTAAAGTTCGTT

GGTTTCGTAACGATCAAGAGGAAACCGCGGGTGTG

GTTAGCACCCCGCTGATCCGTAACGGCGACTGGACC

TTCCAGATTCTGGTTATGCTGGAGATGACCCCGCAA

CGTGGTGATGTGTACACCTGCCACGTTGAACACCCG

AGCCTGCAGAGCCCGATTACCGTGGAGTGGCGTGC

GCAGAGCGAAAGCGCGCAAAGCAAG

34 CD28 TM NT OPT 78 TTTTGGGTTCTGGTGGTTGTGGGTGGCGTGCTGGC

GTGCTACAGCCTGCTGGTGACCGTTGCGTTCATCAT

CTTCTGG

CD28 co- NT OPT 126 | GTGCGTAGCAAACGTAGCCGTCTGCTGCACAGCGA stimulatory CTATATGAACATGACCCCGCGTCGTCCGGGTCCGAC domain CCGTAAGCACTACCAACCGTATGCGCCGCCGCGTGA

CTTTGCGGCGTACCGTAGC

36 CD3 zeta NT OPT 336 | CGTGTTAAATTTAGCCGTAGCGCGGATGCGCCGGC primary GTACCAGCAGGGTCAGAACCAACTGTATAACGAGC signaling TGAACCTGGGCCGTCGTGAGGAATATGACGTGCTG domain GATAAGCGTCGTGGTCGTGATCCGGAAATGGGTGG

CAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCTGT

ACAACGAACTGCAAAAGGACAAAATGGCGGAGGC

GTATAGCGAAATTGGTATGAAGGGCGAGCGTCGTC

GTGGTAAAGGCCACGATGGTCTGTACCAGGGCCTG

AGCACCGCGACCAAAGACACCTATGATGCGCTGCA

CATGCAAGCGCTGCCGCCGCGT

37 viral self- NT OPT 57 GCGACCAACTTCAGCCTGCTGAAGCAGGCGGGTGA cleaving 2A CGTTGAGGAAAACCCGGGCCCG polypeptide 38 Signal NT OPT 69 ATGATCCTGAACAAAGCGCTGATGCTGGGTGCGCT peptide HLA- GGCGCTGACCACCGTTATGAGCCCGTGCGGTGGC

DQA1*05:01 42 HLA- NT OPT 591 | GAGGACATTGTGGCGGATCACGTTGCGAGCTACGG

DQA1*05:01 CGTGAACCTGTACCAGAGCTATGGTCCGAGCGGCC

AATACACCCACGAGTTCGACGGTGATGAACAATTTT

ATGTTGACCTGGGCCGTAAGGAAACCGTGTGGTGC

CTGCCGGTTCTGCGTCAGTTCCGTTTTGATCCGCAAT

TCGCGCTGACCAACATCGCGGTGCTGAAGCACAAC

CTGAACAGCCTGATTAAACGTAGCAACAGCACCGC

GGCGACCAACGAGGTTCCGGAAGTGACCGTTTTCA

GCAAAAGCCCGGTGACCCTGGGTCAGCCGAACATC

CTGATTTGCCTGGTTGACAACATCTTTCCGCCGGTT

GTGAACATTACCTGGCTGAGCAACGGTCACAGCGT

GACCGAGGGCGTTAGCGAAACCAGCTTCCTGAGCA

AGAGCGATCACAGCTTCTTTAAAATCAGCTATCTGA

CCCTGCTGCCGAGCGCGGAGGAAAGCTATGACTGC

AAGGTGGAGCACTGGGGTCTGGATAAGCCGCTGCT

GAAACACTGGGAGCCGGAAATTCCGGCGCCGATGA

GCGAGCTGACCGAAACCGTTGTGTGC

39 CD28 TM NT OPT 78 TTTTGGGTTCTGGTTGTGGTTGGTGGCGTGTTAGCT

TGCTATAGCCTGCTGGTTACCGTGGCGTTTATTATCT

TCTGG

CD28 co- NT OPT 126 | GTTCGCAGCAAGCGTAGCCGTCTGCTGCATAGCGAT stimulatory TACATGAATATGACCCCGCGTCGTCCTGGCCCGACC domain CGCAAACATTATCAACCGTACGCGCCGCCGCGTGAC

TTTGCAGCGTATCGTAGC

41 CD3 zeta NT OPT 336 | CGTGTTAAGTTTAGCCGTAGCGCGGACGCGCCGGC primary GTATCAACAGGGCCAAAATCAGCTGTACAATGAACT signaling GAATCTGGGTCGTCGTGAAGAGTACGATGTTCTGG domain ACAAACGTCGTGGTCGTGACCCGGAGATGGGTGGC

AAACCGCGTCGTAAGAACCCGCAGGAAGGTTTATA

TAATGAGCTGCAGAAAGATAAGATGGCGGAAGCGT

ATAGCGAAATCGGTATGAAGGGCGAACGTCGTCGT

GGCAAGGGTCATGACGGCCTGTATCAAGGTCTGAG

CACCGCGACCAAGGATACCTACGACGCGCTGCATAT

GCAGGCGCTGCCGCCGCGT

Table 3: CHAR HLA-DQ2.5-gliadin-Q-gliadin-a molecule

POLYPEPTIDE Nucleotide (NT) or

Nucleotide Codon

Optimised (NT OPT)

CHAR_HLA- AA 888 | MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGPQQP

DQ2.5-glia- FPQPEQPFPWOPOQGGGGGOLOPFPOPELPYPQPQL

Q102-glia- GSGSGSLGSGSGSGSGSRDSPEDFVYQFKGMCYFTN alo? GTERVRLVSRSIYNREEIVREDSDVGEFRAVTLLGLPAA molecule (Q- EYWNSQKDILERKRAAVDRVCRHNYQLELRTTLQRRY gliadin and EPTVTISPSRTEALNHHNLLVCSVTDFYPAQIKVRWER a-gliadin) NDQEETAGVVSTPLIRNGDWTFQILVMLEMTPQRGD

VYTCHVEHPSLOSPITVEWRAQSESAQSKFWVLVVVG

GVLACYSLLVTVAFIIFFWVRSKRSRLLHSDYMNMTPRR

PGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQ

GQONQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRR

KNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHD

GLYQGLSTATKDTYDALHMOALPPRGSGATNFSLLKQ

AGDVEENPGPMILNKALMLGALALTTVMSPCGGEDI

VADHVASYGVNLYQSYGPSGOYTHEFDGDEQFYVDL

GRKETVWCLPVLROFRFDPOFALTNIAVLKHNLNSLIK

RSNSTAATNEVPEVTVFSKSPVTLGOPNILICLVDNIFP

PVVNITWLSNGHSVTEGVSETSFLSKSDHSFFKISYLTLL

PSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSELTE

TVVCFWVLVVVGGVLACYSLLVTVAFIJFWVRSKRSRL

LHSDYMNMTPRRPGPTRKHYQOPYAPPRDFAAYRSRV

KFSRSADAPAYQQGONQLYNELNLGRREEYDVLDKR

RGRDPEMGGKPRRKNPQEGLYNELOQKDKMAEAYSEI

GMKGERRRGKGHDGLYQGLSTATKDTYDALHMOAL

PPR

2 Signal AA 32 MSWKKALRIPGGLRAATVTLMLSMLSTPVAEG peptide HLA-

DQB1*02:01 5' Flanking AA 3 et 44 HLA-DQ2- AA PFPQPEQPF ee

HLA-DQ2- AA PQPEQPFPW lee |" 46 Complete AA 11 PFPQPEQPFPW gliadin omega peptide

(HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) 3'Flanking AA ee | CT = 5' Flanking AA 3 QLa er | CO 3 HLA-DQ2- AA PFPQPELPY

Cee 4 HLA-DQ2- AA PQPELPYPQ

BEES

Complete AA 11 PFPQPELPYPQ alpha gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2) 3'Flanking AA 3 PQL ee eee] 7 HLA- AA 198 RDSPEDFVYQFKGMCYFTNGTERVRLVSRSIYNREEIV

DQB1*02:01 RFDSDVGEFRAVTLLGLPAAEYWNSQKDILERKRAAV

EC DRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHN

LLVCSVTDFYPAQIKVRWFRNDOEETAGVVSTPLIRN

GDWTFQILVMLEMTPQRGDVYTCHVEHPSLQSPITV

EWRAQSESAQSK

8 (CBM | AM 26 FWVLVVVGGVLACYSLLVTVAFIIFW

CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRD stimulatory FAAYRS domain

CD3 zeta RVKFSRSADAPAYQQGQONQLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAY signaling SEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ domain ALPPR a 11 viral self- AA 19 ATNFSLLKOAGDVEENPGP cleaving 2A polypeptide 12 Signal AA 23 MILNKALMLGALALTTVMSPCGG peptide HLA-

DQA1*05:01 13 HLA- AA 197 EDIVADHVASYGVNLYQSYGPSGQYTHEFDGDEQFYV

DQA1*05:01 DLGRKETVWCLPVLRQFRFDPQFALTNIAVLKHNLNS

LIKRSNSTAATNEVPEVTVFSKSPVTLGOPNILICLVDNI

FPPVVNITWLSNGHSVTEGVSETSFLSKSDHSFEKISYL

TLLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSEL

TETVVC

CD28 co- AA 42 VRSKRSRLLHSDYMNMTPRRPGPTRKHYOPYAPPRD stimulatory FAAYRS domain 10 CD3 zeta RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLD primary KRRGRDPEMGGKPRRKNPQEGLYNELOQKDKMAEAY signaling SEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ, domain ALPPR mw 57 CHAR_HLA- NT WT 2667 | ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG

DQ2.5-glia- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT

Q102-glia- GCTGAGCACCCCGGTGGCGGAGGGTCCACAACAAC ala2 CTTTTCCACAGCCGGAACAACCATTTCCCTGGCAAC molecule (Q- CACAAGGTGGTGGTGGTGGTCAACTTCAACCITTTC gliadin and CTCAACCTGAACTTCCTTATCCTCAACCTCAACTTGG a-gliadin) TAGCGGTAGCGGTAGCCTGGGCAGCGGCAGCGGT

AGCGGTAGCGGCAGCCGTGACAGCCCGGAGGATIT

CGTTTACCAATTCAAGGGCATGTGCTACTTCACCAA

CGGCACCGAACGTGTGCGTCTGGTTAGCCGTAGCA

TCTACAACCGTGAGGAAATTGTGCGTTTCGACAGCG

ATGTTGGCGAGTTTCGTGCGGTGACCCTGCTGGGTC

TGCCGGCGGCGGAGTACTGGAACAGCCAGAAGGA

CATCCTGGAACGTAAACGTGCGGCGGTGGATCGTG

TTTGCCGTCACAACTATCAGCTGGAGCTGCGTACCA

CCCTGCAACGTCGTGTGGAACCGACCGTTACCATCA

GCCCGAGCCGTACCGAAGCGCTGAACCACCACAAC

CTGCTGGTGTGCAGCGTTACCGACTTCTACCCGGCG

CAGATTAAAGTTCGTTGGTTTCGTAACGATCAAGAG

GAAACCGCGGGTGTGGTTAGCACCCCGCTGATCCG

TAACGGCGACTGGACCTTCCAGATTCTGGTTATGCT

GGAGATGACCCCGCAACGTGGTGATGTGTACACCT

GCCACGTTGAACACCCGAGCCTGCAGAGCCCGATT

ACCGTGGAGTGGCGTGCGCAGAGCGAAAGCGCGC

AAAGCAAGTTTTGGGTTCTGGTGGTTGTGGGTGGC

GTGCTGGCGTGCTACAGCCTGCTGGTGACCGTTGC

GTTCATCATCTTCTGGGTGCGTAGCAAACGTAGCCG

TCTGCTGCACAGCGACTATATGAACATGACCCCGCG

TCGTCCGGGTCEGACCCGTAAGCACTACCAACCGTA

TGCGCCGECGCGTGACTTTGCGGCGTACCGTAGCE

GTGTTAAATTTAGCCGTAGCGCGGATGCGCCGGCG

TACCAGCAGGGTCAGAACCAACTGTATAACGAGCT

GAACCTGGGCCGTCGTGAGGAATATGACGTGCTGG

ATAAGCGTCGTGGTCGTGATCCGGAAATGGGTGGC

AAGCCGCGTCGTAAAAACCCGCAGGAAGGTCTGTA

CAACGAACTGCAAAAGGACAAAATGGCGGAGGCGT

ATAGCGAAATTGGTATGAAGGGCGAGCGTCGTCGT

GGTAAAGGCCACGATGGTCTGTACCAGGGCCTGAG

CACCGCGACCAAAGACACCTATGATGCGCTGCACAT

GCAAGCGCTGCCGCCGCGTGGTAGCGGTGCGACCA

ACTTCAGCCTGCTGAAGCAGGCGGGTGACGTTGAG

GAAAACCCGGGCCCGATGATCCTGAACAAAGCGCT

GATGCTGGGTGCGCTGGCGCTGACCACCGTTATGA

GCCCGTGCGGTGGCGAGGACATTGTGGCGGATCAC

GTTGCGAGCTACGGCGTGAACCTGTACCAGAGCTA

TGGTCCGAGCGGCCAATACACCCACGAGTTCGACG

GTGATGAACAATTTTATGTTGACCTGGGCCGTAAGG

AAACCGTGTGGTGCCTGCCGGTTCTGCGTCAGTTCC

GTTTTGATCCGCAATTCGCGCTGACCAACATCGCGG

TGCTGAAGCACAACCTGAACAGCCTGATTAAACGTA

GCAACAGCACCGCGGCGACCAACGAGGTTCCGGAA

GTGACCGTTTTCAGCAAAAGCCCGGTGACCCTGGGT

CAGCCGAACATCCTGATTTGCCTGGTTGACAACATC

TTTCCGCCGGTTGTGAACATTACCTGGCTGAGCAAC

GGTCACAGCGTGACCGAGGGCGTTAGCGAAACCAG

CTTCCTGAGCAAGAGCGATCACAGCTTCTTTAAAAT

CAGCTATCTGACCCTGCTGCCGAGCGCGGAGGAAA

GCTATGACTGCAAGGTGGAGCACTGGGGTCTGGAT

AAGCCGCTGCTGAAACACTGGGAGCCGGAAATTCC

GGCGCCGATGAGCGAGCTGACCGAAACCGTTGTGT

GCTTTTGGGTTCTGGTTGTGGTTGGTGGCGTGTTAG

CTTGCTATAGCCTGCTGGTTACCGTGGCGTTTATTAT

CTTCTGGGTTCGCAGCAAGCGTAGCCGTCTGCTGCA

TAGCGATTACATGAATATGACCCCGCGTCGTCCTGG

CCCGACCCGCAAACATTATCAACCGTACGCGCCGCC

GCGTGACTTTGCAGCGTATCGTAGCCGTGTTAAGTT

TAGCCGTAGCGCGGACGCGCCGGCGTATCAACAGG

GCCAAAATCAGCTGTACAATGAACTGAATCTGGGTC

GTCGTGAAGAGTACGATGTTCTGGACAAACGTCGT

GGTCGTGACCCGGAGATGGGTGGCAAACCGCGTCG

TAAGAACCCGCAGGAAGGTTTATATAATGAGCTGC

AGAAAGATAAGATGGCGGAAGCGTATAGCGAAATC

GGTATGAAGGGCGAACGTCGTCGTGGCAAGGGTCA

TGACGGCCTGTATCAAGGTCTGAGCACCGCGACCA

AGGATACCTACGACGCGCTGCATATGCAGGCGCTG

CCGCCGCGTTAA

DQB1*02:01 GCTGAGCACCCCAGTGGCTGAGGGC 5' Flanking NT WT CCACAACAA eee | TT 48 HLA-DQ2- CCTTTTCCACAGCCGGAACAACCATTT en 49 HLA-DQ2- 27 CCACAGCCGGAACAACCATTTCCCTGG lea

Complete NT WT 33 CCTTTTCCACAGCCGGAACAACCATTTCCCTGG gliadin omega peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-02) 3'Flanking NT WT CAACCACAA

A

5' Flanking NT WT CAACTTCAA ee TT 16 HLA-DQ2- NT WT 26 CCTTTTCCTCAACCTGAACTTCCTTA

Cen | Tm 17 HLA-DQ2- CCTCAACCTGAACTTCCTTATCCTCAA laa EE 18 Complete NT WT CCTTTTCCTCAACCTGAACTTCCTTATCCTCAA alpha gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2)

3'Flanking NT WT 9 CCTCAACTT oe 19 Linker NT WT 51 GGCTCTGGATCTGGGTCCCTGGGATCTGGCTCTGG

Te

HLA- NT WT 594 | AGAGACTCTCCCGAGGATTTCGTGTACCAGTTTAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGGACAGAGCGCGT

EC GCGTCTTGTGAGCAGAAGCATCTATAACCGAGAAG

AGATCGTGCGCTTCGACAGCGACGTGGGGGAGTTC

CGGGCGGTGACGCTGCTGGGGCTGCCTGCCGCCGA

GTACTGGAACAGCCAGAAGGACATCCTGGAGAGGA

AACGGGCGGCGGTGGACAGGGTGTGCAGACACAA

CTACCAGTTGGAGCTCCGCACGACCTTGCAGCGGC

GAGTGGAGCCCACAGTGACCATCTCCCCATCCAGG

ACAGAGGCCCTCAACCACCACAACCTGCTGGTCTGC

TCGGTGACAGATTTCTATCCAGCCCAGATCAAAGTC

CGGTGGTTTCGGAATGACCAGGAGGAGACAGCTGG

CGTTGTGTCCACCCCCCTTATTAGGAATGGTGACTG

GACCTTCCAGATCCTGGTGATGCTGGAAATGACTCC

CCAGCGTGGAGACGTCTACACCTGCCACGTGGAGC

ACCCCAGCCTCCAGAGCCCCATCACCGTGGAGTGGC

GGGCTCAATCTGAATCTGCCCAGAGCAAG

21 CD28 TM NT WT 78 TTTTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATT

TTCTGG

ACTTCGCAGCCTATCGCTCC

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAG

GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCG

GAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTC

TCAGTACAGCCACCAAGGACACCTACGACGCCCTTC

ACATGCAGGCCCTGCCCCCTCGC

Re 24 viral self- NT WT 57 GCCACCAACTTCAGCCTGCTGAAGCAGGCCGGCGA cleaving 2A CGTGGAGGAAAACCCTGGGCCC polypeptide

DQA1*05:01 26 HLA- NT WT 591 | GAAGACATTGTGGCTGACCACGTCGCCTCTTATGGT

DQA1*05:01 GTAAACTTGTACCAGTCTTACGGTCCCTCTGGCCAG

TACACCCATGAATTTGATGGAGATGAGCAGTTCTAC

GTGGACCTGGGGAGGAAGGAGACTGTCTGGTGTTT

GCCTGTTCTCAGACAATTTAGATTTGACCCGCAATTT

GCACTGACAAACATCGCTGTCCTAAAACATAACTTG

AACAGTCTGATTAAACGCTCCAACTCTACCGCTGCT

ACCAATGAGGTTCCTGAGGTCACAGTGTTTTCCAAG

TCTCCCGTGACACTGGGTCAGCCCAACATCCTCATCT

GTCTTGTGGACAACATCTTTCCTCCTGTGGTCAACAT

CACATGGCTGAGCAATGGGCACTCAGTCACAGAAG

GTGTTTCTGAGACCAGCTTCCTCTCCAAGAGTGATC

ATTCCTTCTTCAAGATCAGTTACCTCACCCTCCTCCCT

TCTGCTGAGGAGAGTTATGACTGCAAGGTGGAGCA

CTGGGGCCTGGACAAGCCTCTTCTGAAACACTGGG

AGCCTGAGATTCCAGCCCCTATGTCAGAGCTCACAG

AGACTGTGGTCTGC

21 CD28 TM NT WT 78 TITTGGGTGCTGGTGGTGGTTGGGGGAGTCCTGGC

TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATIATT

TTCTGG

22 CD28 co- NT WT 126 GTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTG stimulatory ACTACATGAACATGACTCCCCGCCGCCCCGGGCCCA domain

CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCG

Mn 23 CD3 zeta NT WT 336 | AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGC primary GTACCAGCAGGGCCAGAACCAGCTCTATAACGAGC signaling TCAATCTAGGACGAAGAGAGGAGTACGATGTTTTG domain GACAAGAGACGTGGCCGGGACCCTGAGATGGGGG

GAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCT

GTACAATGAACTGCAGAAAGATAAGATGGCGGAG

GCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCG

GAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTC

TCAGTACAGCCACCAAGGACACCTACGACGCCCTTC

ACATGCAGGCCCTGCCCCCTCGC

59 CHAR_HLA- NT OPT 2667 | ATGAGCTGGAAGAAAGCGCTGCGTATCCCGGGTGG

DQ2.5-glia- CCTGCGTGCGGCGACCGTTACCCTGATGCTGAGCAT

Q102-glia- GCTGAGCACCCCGGTGGCGGAGGGTCCACAGCAGC ala2 CCTTCCCTCAGCCAGAGCAGCCCTTTCCTTGGCAGC molecule (Q- CCCAGGGCGGCGGCGGCGGCCAGCTGCAACCGTIT gliadin and CCGCAGCCGGAACTGCCGTACCCGCAGCCGCAACT a-gliadin) GGGTAGCGGTAGCGGTAGCCTGGGCAGCGGCAGC

GGTAGCGGTAGCGGCAGCCGTGACAGCCCGGAGG

ATTTCGTTTACCAATTCAAGGGCATGTGCTACTTCAC

CAACGGCACCGAACGTGTGCGTCTGGTTAGCCGTA

GCATCTACAACCGTGAGGAAATTGTGCGTTTCGACA

GCGATGTTGGCGAGTTTCGTGCGGTGACCCTGCTG

GGTCTGCCGGCGGCGGAGTACTGGAACAGCCAGAA

GGACATCCTGGAACGTAAACGTGCGGCGGTGGATC

GTGTITGCCGTCACAACTATCAGCTGGAGCTGCGTA

CCACCCTGCAACGTCGTGTGGAACCGACCGTTACCA

TCAGCCCGAGCCGTACCGAAGCGCTGAACCACCAC

AACCTGCTGGTGTGCAGCGTTACCGACTTCTACCCG

GCGCAGATTAAAGTTCGTTGGTTTCGTAACGATCAA

GAGGAAACCGCGGGTGTGGTTAGCACCCCGCTGAT

CCGTAACGGCGACTGGACCTTCCAGATTCTGGTTAT

GCTGGAGATGACCCCGCAACGTGGTGATGTGTACA

CCTGCCACGTTGAACACCCGAGCCTGCAGAGCCEG

ATTACCGTGGAGTGGCGTGCGCAGAGCGAAAGCGC

GCAAAGCAAGTTTTGGGTTCTGGTGGTTGTGGGTG

GCGTGCTGGCGTGCTACAGCCTGCTGGTGACCGTT

GCGTTCATCATCTTCTGGGTGCGTAGCAAACGTAGC

CGTCTGCTGCACAGCGACTATATGAACATGACCCCG

CGTCGTCCGGGTCCGACCCGTAAGCACTACCAACCG

TATGCGCCGCCGCGTGACTTTGCGGCGTACCGTAGC

CGTGTTAAATTTAGCCGTAGCGCGGATGCGCCGGC

GTACCAGCAGGGTCAGAACCAACTGTATAACGAGC

TGAACCTGGGCCGTCGTGAGGAATATGACGTGCTG

GATAAGCGTCGTGGTCGTGATCCGGAAATGGGTGG

CAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCTGT

ACAACGAACTGCAAAAGGACAAAATGGCGGAGGC

GTATAGCGAAATTGGTATGAAGGGCGAGCGTCGTC

GTGGTAAAGGCCACGATGGTCTGTACCAGGGCCTG

AGCACCGCGACCAAAGACACCTATGATGCGCTGCA

CATGCAAGCGCTGCCGCCGCGTGGTAGCGGTGCGA

CCAACTTCAGCCTGCTGAAGCAGGCGGGTGACGTT

GAGGAAAACCCGGGCCCGATGATCCTGAACAAAGC

GCTGATGCTGGGTGCGCTGGCGCTGACCACCGTTAT

GAGCCCGTGCGGTGGCGAGGACATTGTGGCGGATC

ACGTTGCGAGCTACGGCGTGAACCTGTACCAGAGC

TATGGTCCGAGCGGCCAATACACCCACGAGTTCGAC

GGTGATGAACAATTTTATGTTGACCTGGGCCGTAAG

GAAACCGTGTGGTGCCTGCCGGTTCTGCGTCAGTTC

CGTTTTGATCCGCAATTCGCGCTGACCAACATCGCG

GTGCTGAAGCACAACCTGAACAGCCTGATTAAACGT

AGCAACAGCACCGCGGCGACCAACGAGGTTCCGGA

AGTGACCGTTTTCAGCAAAAGCCCGGTGACCCTGG

GTCAGCCGAACATCCTGATTTGCCTGGTTGACAACA

TCTTTCCGCCGGTTGTGAACATTACCTGGCTGAGCA

ACGGTCACAGCGTGACCGAGGGCGTTAGCGAAACC

AGCTTCCTGAGCAAGAGCGATCACAGCTTCTTTAAA

ATCAGCTATCTGACCCTGCTGCCGAGCGCGGAGGA

AAGCTATGACTGCAAGGTGGAGCACTGGGGTCTGG

ATAAGCCGCTGCTGAAACACTGGGAGCCGGAAATT

CCGGCGCCGATGAGCGAGCTGACCGAAACCGTTGT

GTGCTTTTGGGTTCTGGTTGTGGTTGGTGGCGTGTT

AGCTTGCTATAGCCTGCTGGTTACCGTGGCGTTTAT

TATCTTCTGGGTTCGCAGCAAGCGTAGCCGTCTGCT

GCATAGCGATTACATGAATATGACCCCGCGTCGTCC

TGGCCCGACCCGCAAACATTATCAACCGTACGCGCC

GCCGCGTGACTTTGCAGCGTATCGTAGCCGTGTTAA

GTTTAGCCGTAGCGCGGACGCGCCGGCGTATCAAC

AGGGCCAAAATCAGCTGTACAATGAACTGAATCTG

GGTCGTCGTGAAGAGTACGATGTTCTGGACAAACG

TCGTGGTCGTGACCCGGAGATGGGTGGCAAACCGC

GTCGTAAGAACCCGCAGGAAGGTTTATATAATGAG

CTGCAGAAAGATAAGATGGCGGAAGCGTATAGCGA

AATCGGTATGAAGGGCGAACGTCGTCGTGGCAAGG

GTCATGACGGCCTGTATCAAGGTCTGAGCACCGCG

ACCAAGGATACCTACGACGCGCTGCATATGCAGGC

GCTGCCGCCGCGTTAA

DQB1*02:01 GCTGAGCACCCCGGTGGCGGAGGGT 5' Flanking NT OPT CCACAACAA et | TTC 52 HLA-DQ2- NT OPT 27 CCTTTTCCACAGCCGGAACAACCATTT lea TT 53 HLA-DQ2- NT OPT 27 CCACAGCCGGAACAACCATTTCCCTGG lee 54 Complete NT OPT 33 CCTTTTCCACAGCCGGAACAACCATTTCCCTGG gliadin omega peptide

(HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) 3'Flanking NT OPT CAACCACAA en 5' Flanking NT OPT CAGCTGCAA

IEE a 29 HLA-DQ2- NT OPT 27 CCGTTTCCGCAGCCGGAACTGCCGTAC

Ce | TTT

HLA-DQ2- NT OPT 27 CCGCAGCCGGAACTGCCGTACCCGCAG

Cea | TT 31 Complete NT OPT 33 CCGTTTCCGCAGCCGGAACTGCCGTACCCGCAG alpha gliadin peptide (HLA-DQ2- glia-ala and

HLA-DQ2- glia-a2) 3'Flanking NT OPT CCGCAACTG le | TT 32 Linker NT OPT 51 GGTAGCGGTAGCGGTAGCCTGGGCAGCGGCAGCG ee 33 HLA- NT OPT 594 CGTGACAGCCCGGAGGATTTCGTTTACCAATTCAAG

DQB1*02:01 GGCATGTGCTACTTCACCAACGGCACCGAACGTGTG

EC CGTCTGGTTAGCCGTAGCATCTACAACCGTGAGGAA

ATTGTGCGTTTCGACAGCGATGTTGGCGAGTTICGT

GCGGTGACCCTGCTGGGTCTGCCGGCGGCGGAGTA

CTGGAACAGCCAGAAGGACATCCTGGAACGTAAAC

GTGCGGCGGTGGATCGTGTTTGCCGTCACAACTATC

AGCTGGAGCTGCGTACCACCCTGCAACGTCGTGTG

GAACCGACCGTTACCATCAGCCCGAGCCGTACCGA

AGCGCTGAACCACCACAACCTGCTGGTGTGCAGCG

TTACCGACTTCTACCCGGCGCAGATTAAAGTTCGTT

GGTTTCGTAACGATCAAGAGGAAACCGCGGGTGTG

GTTAGCACCCCGCTGATCCGTAACGGCGACTGGACC

TTCCAGATTCTGGTTATGCTGGAGATGACCCCGCAA

CGTGGTGATGTGTACACCTGCCACGTTGAACACCCG

AGCCTGCAGAGCCCGATTACCGTGGAGTGGCGTGC

GCAGAGCGAAAGCGCGCAAAGCAAG

34 CD28 TM NT OPT 78 TTTTGGGTTCTGGTGGTTGTGGGTGGCGTGCTGGC

GTGCTACAGCCTGCTGGTGACCGTTGCGTTCATCAT

CTTCTGG

CTTTGCGGCGTACCGTAGC

CAAGCCGCGTCGTAAAAACCCGCAGGAAGGTCTGT

ACAACGAACTGCAAAAGGACAAAATGGCGGAGGC

GTATAGCGAAATTGGTATGAAGGGCGAGCGTCGTC

GTGGTAAAGGCCACGATGGTCTGTACCAGGGCCTG

AGCACCGCGACCAAAGACACCTATGATGCGCTGCA

CATGCAAGCGCTGCCGCCGCGT

DQA1*05:01 42 HLA- NT OPT 591 GAGGACATTGTGGCGGATCACGTTGCGAGCTACGG

DQA1*05:01 CGTGAACCTGTACCAGAGCTATGGTCCGAGCGGCC

AATACACCCACGAGTTCGACGGTGATGAACAATTTT

ATGTTGACCTGGGCCGTAAGGAAACCGTGTGGTGC

CTGCCGGTTCTGCGTCAGTTCCGTTTTGATCCGCAAT

TCGCGCTGACCAACATCGCGGTGCTGAAGCACAAC

CTGAACAGCCTGATTAAACGTAGCAACAGCACCGC

GGCGACCAACGAGGTTCCGGAAGTGACCGTTTTCA

GCAAAAGCCCGGTGACCCTGGGTCAGCCGAACATC

CTGATTTGCCTGGTTGACAACATCTTTCCGCCGGTT

GTGAACATTACCTGGCTGAGCAACGGTCACAGCGT

GACCGAGGGCGTTAGCGAAACCAGCTTCCTGAGCA

AGAGCGATCACAGCTTCTTTAAAATCAGCTATCTGA

CCCTGECTGCCGAGCGCGGAGGAAAGCTATGACTGC

AAGGTGGAGCACTGGGGTCTGGATAAGCCGCTGCT

GAAACACTGGGAGCCGGAAATTCCGGCGCCGATGA

GCGAGCTGACCGAAACCGTTGTGTGC

39 CD28 TM NT OPT 78 TITTGGGTICTGGTTGTGGTIGGTGGCGTGTTAGCT

TGCTATAGCCTGCTGGTTACCGTGGCGTTTATTATCT

TCTGG

40 CD28 co- NT OPT 126 GTTCGCAGCAAGCGTAGCCGTCTGCTGCATAGCGAT stimulatory TACATGAATATGACCCCGCGTECGTCCTGGCCCGACC domain CGCAAACATTATCAACCGTACGCGCCGCCGCGTGAC

TTTGCAGCGTATCGTAGC

41 CD3 zeta 336 CGTGTTAAGTITAGCCGTAGCGCGGACGCGCCGGC primary GTATCAACAGGGCCAAAATCAGCTGTACAATGAACT signaling GAATCTGGGTCGTCGTGAAGAGTACGATGTTCTGG domain ACAAACGTCGTGGTCGTGACCCGGAGATGGGTGGC

AAACCGCGTCGTAAGAACCCGCAGGAAGGTTTATA

TAATGAGCTGCAGAAAGATAAGATGGCGGAAGCGT

ATAGCGAAATCGGTATGAAGGGCGAACGTCGTCGT

GGCAAGGGTCATGACGGCCTGTATCAAGGTCTGAG

CACCGCGACCAAGGATACCTACGACGCGCTGCATAT

GCAGGCGCTGCCGCCGCGT

Te

Table 4: Additional Sequences u =

POLYPEPTIDE Nucleotide (NT) or

Nucleotide Codon

Optimised (NT OPT) 61 HLA-DQ8- AA 9 | EGSFQPSQE me | CT 62 CLIP NT 28 | CCGCTGCTGATGCAGGCGCTGCCGATG ee 63 Complete AA 14 QLOPFPOPELPYPQ alpha gliadin peptide (HLA-DQ2- glia-ala and HLA-

DQ2-glia- a2) with 5’ flanking 64 Complete AA 17 QLOPFPQPELPYPQPQL alpha gliadin peptide (HLA-DQ2- glia-ala and HLA-

DQ2-glia- a2) with 5’ flanking and 3’ flanking 65 Complete AA 14 PFPQPELPYPQPQL alpha gliadin peptide

(HLA-DQ2- glia-ala and HLA-

DQ2-glia- 02) with 3’ flanking

Complete AA 14 PQQPFPQPEOQPFPW gliadin omega peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) with 5’ flanking 67 Complete AA 17 PQQOPFPQPEQPFPWQPQ gliadin omega peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) with 5’ flanking and 3’ flanking

Complete AA 14 PFPQPEQPFPWQPQ gliadin omega peptide (HLA-DQ2- glia-Q1 and

HLA-DQ2- glia-Q2) with 3’ flanking

HLA-DQ2- AA PYPQPELPY

El I I 70 Complete AA PYPQPELPYPQ gliadin alpha peptide (HLA-DQ2- glia-alb and HLA-

DQ2-glia- a2) 71 Complete QLQPYPQPELPYPQ alpha gliadin peptide (HLA-DQ2- glia-alb and HLA-

DQ2-glia- 02) with 5’ flanking 72 Complete QLQPYPQPELPYPQPQL alpha gliadin peptide (HLA-DQ2- glia-a1b and HLA-

DQ2-glia- 02) with 5’

flanking and 3’ flanking 73 Complete PYPQPELPYPQPQL alpha gliadin peptide (HLA-DQ2- glia-alb and HLA-

DQ2-glia- a2) with 3’ flanking

References 1. Horn, P.A., et al., Highly efficient gene transfer into baboon marrow repopulating cells using GALV-pseudotype oncoretroviral vectors produced by human packaging cells.

Blood, 2002. 100(12): p. 3960-3967. 2. Petersen, J., et al, T-cell receptor recognition of HLA-DQ2-gliadin complexes associated with celiac disease. Nature Structural & Molecular Biology, 2014. 21(5): p. 480-488. 3. Vader, W., et al., The gluten response in children with celiac disease is directed toward multiple gliadin and glutenin peptides. Gastroenterology, 2002. 122(7): p. 1729-1737.

NL P366532NL Academisch Ziekenhuis Leiden (h.o.d.n. LUMC) Agents for treating

Celiac Disease 73 866 AA PAT source 1..866 mol_type protein organism synthetic construct REGION 1..866 note CHAR_HLA-DQ2.5-glia-alphala-glia-alpha2 molecule (alpha-gliadin)

MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGQLQPFPQPELPYPQPQLGSGSGSLGSGSGS

GSGSRDSPEDFVYQFKGMCYFTNGTERVRLVSRSIYNREEIVRFDSDVGEFRAVTLLGLPAAE

YWNSQKDILERKRAAVDRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHNLLVCSVTDF

YPAQIKVRWFRNDQEETAGVVSTPLIRNGDWTFQILVMLEMTPQRGDVYTCHVEHPSLQSPIT

VEWRAQSESAQSKFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGP

TRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDP

EMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDAL

HMQALPPRGSGATNFSLLKQAGDVEENPGPMILNKALMLGALALTTVMSPCGGEDIVADHVA

SYGVNLYQSYGPSGQYTHEFDGDEQFYVDLGRKETVWCLPVLRQFRFDPQFALTNIAVLKHNL

NSLIKRSNSTAATNEVPEVTVFSKSPVTLGQPNILICLVDNIFPPVVNITWLSNGHSVTEGVSET

SFLSKSDHSFFKISYLTLLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSELTETVVCFWVL

VVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYR

SRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYN

ELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR 32 AA PAT source 1..32 mol_type protein organism Homo sapiens

MSWKKALRIPGGLRAATVTLMLSMLSTPVAEG 9 AA PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note HLA-DQ2-glia-alphala PFPQPELPY 9 AA

PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note HLA-

DQ2-glia-alpha2 PQPELPYPQ 11 AA PAT source 1..11 mol_type protein organism unidentified REGION 1..11 note Complete alpha gliadin peptide (HLA-DQ2-glia- alphala and HLA-DQ2-glia-alpha2) PFPQPELPYPQ 17 AA PAT source 1..17 mol_type protein organism synthetic construct REGION 1..17 note Linker

GSGSGSLGSGSGSGSGS 198 AA PAT source 1..198 mol_type protein organism

Homo sapiens

RDSPEDFVYQFKGMCYFTNGTERVRLVSRSIYNREEIVRFDSDVGEFRAVTLLGLPAAEYWNS

QKDILERKRAAVDRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHNLLVCSVTDFYPAQI

KVRWFRNDQEETAGVVSTPLIRNGDWTFQILVMLEMTPQRGDVYTCHVEHPSLQSPITVEWR

AQSESAQSK 26 AA PAT source 1..26 mol_type protein organism Homo sapiens

FWVLVVVGGVLACYSLLVTVAFIIFW 42 AA PAT source 1..42 mol_type protein organism Homo sapiens VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 112 AA PAT source 1..112 mol_type protein organism Homo sapiens

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNE

LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR 19 AA PAT source 1..19 mol_type protein organism porcine teschovirus-1 2A

ATNFSLLKQAGDVEENPGP 23 AA PAT source 1..23 mol_type protein organism Homo sapiens MILNKALMLGALALTTVMSPCGG 197 AA PAT source 1..197 mol_type protein organism Homo sapiens

EDIVADHVASYGVNLYQSYGPSGQYTHEFDGDEQFYVDLGRKETVWCLPVLRQFRFDPQFAL

TNIAVLKHNLNSLIKRSNSTAATNEVPEVTVFSKSPVTLGQPNILICLVDNIFPPVVNITWLSNG

HSVTEGVSETSFLSKSDHSFFKISYLTLLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSEL

TETVVC 2601 DNA PAT source 1..2601 mol_type other DNA organism synthetic construct misc_feature 1..2601 note CHAR_HLA-DQ?2.5-glia-alphala-glia-alpha2 molecule (alpha-gliadin) atgtcttggaaaaaggctttgcggatccccggaggccttcgggcagcaactgtgaccttgatgctgtcgatgctgagc accccagtggctgagggccaacttcaaccttttcctcaacctgaacttccttatcctcaacctcaacttggctctggatctg ggtccctgggatctggctctggatctggctctggatctagagactctcccgaggatttcgtgtaccagtttaagggcatg tgctacttcaccaacgggacagagcgcgtgcgtcttgtgagcagaagcatctataaccgagaagagatcgtgcgcttc gacagcgacgtgggggagttccgggcggtgacgctgctggggctgcctgccgccgagtactggaacagccagaag gacatcctggagaggaaacgggcggcggtggacagggtgtgcagacacaactaccagttggagctccgcacgacc ttgcagcggcgagtggagcccacagtgaccatctccccatccaggacagaggccctcaaccaccacaacctgctggt ctgctcggtgacagatttctatccagcccagatcaaagtccggtggtttcggaatgaccaggaggagacagctggcgt tgtgtccaccccccttattaggaatggtgactggaccttccagatcctggtgatgctggaaatgactccccagcgtgga gacgtctacacctgccacgtggagcaccccagcctccagagccccatcaccgtggagtggcgggctcaatctgaatct gcccagagcaagttttgggtgctggtggtggttgggggagtcctggcttgctatagcttgctagtaacagtggcctttat tattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcc cacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtgaagttcagcaggag cgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagta cgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaag gcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccgga ggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcag gccctgccccctcgcggcagcggcgccaccaacttcagcctgctgaagcaggccggcgacgtggaggaaaaccctg ggcccatgatcctaaacaaagctctgatgctgggggcccttgccctgaccaccgtgatgagcccctgtggaggtgaag acattgtggctgaccacgtcgcctcttatggtgtaaacttgtaccagtcttacggtccctctggccagtacacccatgaat ttgatggagatgagcagttctacgtggacctggggaggaaggagactgtctggtgtttgcctgttctcagacaatttag atttgacccgcaatttgcactgacaaacatcgctgtcctaaaacataacttgaacagtctgattaaacgctccaactcta ccgctgctaccaatgaggttcctgaggtcacagtgttttccaagtctcccgtgacactgggtcagcccaacatcctcatc tgtcttgtggacaacatctttcctcctgtggtcaacatcacatggctgagcaatgggcactcagtcacagaaggtgtttct gagaccagcttcctctccaagagtgatcattccttcttcaagatcagttacctcaccctcetcccttctgctgaggagagtt atgactgcaaggtggagcactggggcctggacaagcctcttctgaaacactgggagcctgagattccagcccctatgt cagagctcacagagactgtggtctgcttttgggtgctggtggtggttgggggagtcctggcttgctatagcttgctagta acagtggcctttattattttctgggtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactcccc gccgccccgggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctccagagtga agttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacg aagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaaga accctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaag gcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacg cccttcacatgcaggccctgccccctcgctga 96 DNA PAT source 1..96 mol_type genomic DNA organism Homo sapiens atgtcttggaaaaaggctttgcggatccccggaggccttcgggcagcaactgtgaccttgatgctgtcgatgctgagc accccagtggctgagggc 26 DNA PAT source 1..26 mol_type unassigned DNA organism unidentified misc_feature 1..26 note HLA-DQ2-glia-alphala ccttttcctcaacctgaacttcctta 27 DNA PAT source 1..27 mol_type unassigned DNA organism unidentified misc_feature 1..27 note HLA-DQ2-glia-alpha2 cctcaacctgaacttccttatcctcaa 33 DNA PAT source 1..33 mol_type unassigned DNA organism unidentified misc_feature 1..33 note Complete gliadin peptide (HLA-DQ2- glia-alphala and HLA-DQ2-glia-alpha2) ccttttcctcaacctgaacttccttatcctcaa 51 DNA

PAT source 1..51 mol_type other DNA organism synthetic construct misc_feature 1..51 note Linker ggctctggatctgggtccctgggatctggctctggatctggctctggatct 594 DNA

PAT source 1..594 mol_type genomic DNA organism Homo sapiens agagactctcccgaggatttcgtgtaccagtttaagggcatgtgctacttcaccaacgggacagagcgcgtgcgtcttg tgagcagaagcatctataaccgagaagagatcgtgcgcttcgacagcgacgtgggggagttccgggcggtgacgct gctggggctgcctgccgccgagtactggaacagccagaaggacatcctggagaggaaacgggcggcggtggaca gggtgtgcagacacaactaccagttggagctccgcacgaccttgcagcggcgagtggagcccacagtgaccatctcc ccatccaggacagaggccctcaaccaccacaacctgctggtctgctcggtgacagatttctatccagcccagatcaaa gtccggtggtttcggaatgaccaggaggagacagctggcgttgtgtccaccccccttattaggaatggtgactggacc ttccagatcctggtgatgctggaaatgactccccagcgtggagacgtctacacctgccacgtggagcaccccagcctc cagagccccatcaccgtggagtggcgggctcaatctgaatctgcccagagcaag 78 DNA PAT source 1..78 mol_type genomic DNA organism Homo sapiens ttttgggtgctggtggtggttgggggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgg 126

DNA PAT source 1..126 mol_type genomic DNA organism Homo sapiens gtgaggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccgggcccacccgca agcattaccagccctatgccccaccacgcgacttcgcagcctatcgctcc 336 DNA PAT source 1..336 mol_type genomic DNA organism Homo sapiens agagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatc taggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgaga aggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattggg atgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacc tacgacgcccttcacatgcaggccctgccccctcgc 57 DNA PAT source 1..57 mol_type other

DNA organism synthetic construct misc_feature 1..57 note viral self-cleaving 2A polypeptide gccaccaacttcagcctgctgaagcaggccggcgacgtggaggaaaaccctgggccc 69 DNA

PAT source 1..69 mol_type genomic DNA organism Homo sapiens atgatcctaaacaaagctctgatgctgggggcccttgccctgaccaccgtgatgagcccctgtggaggt 591 DNA

PAT source 1..591 mol_type genomic DNA organism Homo sapiens gaagacattgtggctgaccacgtcgcctcttatggtgtaaacttgtaccagtcttacggtccctctggccagtacaccca tgaatttgatggagatgagcagttctacgtggacctggggaggaaggagactgtctggtgtttgcctgttctcagaca atttagatttgacccgcaatttgcactgacaaacatcgctgtcctaaaacataacttgaacagtctgattaaacgctcca actctaccgctgctaccaatgaggttcctgaggtcacagtgttttccaagtctcccgtgacactgggtcagcccaacatc ctcatctgtcttgtggacaacatctttcctcctgtggtcaacatcacatggctgagcaatgggcactcagtcacagaagg tgtttctgagaccagcttcctctccaagagtgatcattccttcttcaagatcagttacctcaccctcctcccttctgctgagg agagttatgactgcaaggtggagcactggggcctggacaagcctcttctgaaacactgggagcctgagattccagcc cctatgtcagagctcacagagactgtggtctgc 2601 DNA PAT source 1..2601 mol_type other

DNA organism synthetic construct misc_feature 1..2601 note CHAR_HLA-DQ2.5- glia-alphala-glia-alpha2 molecule (alpha-gliadin) atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggtcagctgcaaccgtttccgcagccggaactgccgtacccgcagccgcaactgggtagc ggtagcggtagcctgggcagcggcagcggtagcggtagcggcagccgtgacagcccggaggatttcgtttaccaatt caagggcatgtgctacttcaccaacggcaccgaacgtgtgcgtctggttagccgtagcatctacaaccgtgaggaaat tgtgcgtttcgacagcgatgttggcgagtttcgtgcggtgaccctgctgggtctgccggcggcggagtactggaacag ccagaaggacatcctggaacgtaaacgtgcggcggtggatcgtgtttgccgtcacaactatcagctggagctgcgtac caccctgcaacgtcgtgtggaaccgaccgttaccatcagcccgagccgtaccgaagcgctgaaccaccacaacctgct ggtgtgcagcgttaccgacttctacccggcgcagattaaagttcgttggtttcgtaacgatcaagaggaaaccgcggg tgtggttagcaccccgctgatccgtaacggcgactggaccttccagattctggttatgctggagatgaccccgcaacgt ggtgatgtgtacacctgccacgttgaacacccgagcctgcagagcccgattaccgtggagtggcgtgcgcagagcga aagcgcgcaaagcaagttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgc gttcatcatcttctgggtgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccg ggtccgacccgtaagcactaccaaccgtatgcgccgccgcgtgactttgcggcgtaccgtagccgtgttaaatttagcc gtagcgcggatgcgccggcgtaccagcagggtcagaaccaactgtataacgagctgaacctgggccgtcgtgagga atatgacgtgctggataagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgtaaaaacccgcaggaa ggtctgtacaacgaactgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatgaagggcgagcgtcgtc gtggtaaaggccacgatggtctgtaccagggcctgagcaccgcgaccaaagacacctatgatgcgctgcacatgcaa gcgctgccgccgcgtggtagcggtgcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgg gcccgatgatcctgaacaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggcgag gacattgtggcggatcacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacaccca cgagttcgacggtgatgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgtcag ttccgttttgatccgcaattcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgtagca acagcaccgcggcgaccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagccgaa catcctgatttgcctggttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtgaccg agggcgttagcgaaaccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgccga gcgcggaggaaagctatgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagccgg aaattccggcgccgatgagcgagctgaccgaaaccgttgtgtgcttttgggttctggttgtggttggtggcgtgttagct tgctatagcctgctggttaccgtggcgtttattatcttctgggttcgcagcaagcgtagccgtctgctgcatagcgattac atgaatatgaccccgcgtcgtcctggcccgacccgcaaacattatcaaccgtacgcgccgccgcgtgactttgcagcgt atcgtagccgtgttaagtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaa ctgaatctgggtcgtcgtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaacc gcgtcgtaagaacccgcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatc ggtatgaagggcgaacgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggata cctacgacgcgctgcatatgcaggcgctgccgccgcgttaa 96 DNA PAT source 1..96 mol_type other DNA organism synthetic construct misc_feature 1..96 note Signal peptide

HLA-DQB1*02:01 atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggt 27 DNA PAT source 1..27 mol_type other DNA organism synthetic construct misc_feature 1..27 note HLA-DQ2-glia-alphala ccgtttccgcagccggaactgccgtac 27 DNA PAT source 1..27 mol_type other DNA organism synthetic construct misc_feature 1..27 note HLA-DQ2-glia-alpha2 ccgcagccggaactgccgtacccgcag 33 DNA PAT source 1..33 mol_type other DNA organism synthetic construct misc_feature 1..33 note Complete gliadin peptide (HLA-DQ2-glia-alphala and HLA-DQ2-glia-alpha2) ccgtttccgcagccggaactgccgtacccgcag 51 DNA PAT source 1..51 mol_type other DNA organism synthetic construct misc_feature 1..51 note Linker ggtagcggtagcggtagcctgggcagcggcagcggtagcggtagcggcagc 594 DNA PAT source 1..594 mol_type other DNA organism synthetic construct misc_feature 1..594 note

HLA-DQB1*02:01 cgtgacagcccggaggatttcgtttaccaattcaagggcatgtgctacttcaccaacggcaccgaacgtgtgcgtctgg ttagccgtagcatctacaaccgtgaggaaattgtgcgtttcgacagcgatgttggcgagtttcgtgcggtgaccctgct gggtctgccggcggcggagtactggaacagccagaaggacatcctggaacgtaaacgtgcggcggtggatcgtgtt tgccgtcacaactatcagctggagctgcgtaccaccctgcaacgtcgtgtggaaccgaccgttaccatcagcccgagc cgtaccgaagcgctgaaccaccacaacctgctggtgtgcagcgttaccgacttctacccggcgcagattaaagttcgtt ggtttcgtaacgatcaagaggaaaccgcgggtgtggttagcaccccgctgatccgtaacggcgactggaccttccag attctggttatgctggagatgaccccgcaacgtggtgatgtgtacacctgccacgttgaacacccgagcctgcagagc ccgattaccgtggagtggcgtgcgcagagcgaaagcgcgcaaagcaag 78 DNA PAT source 1..78 mol_type other DNA organism synthetic construct misc_feature 1..78 note CD28 TM ttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgcgttcatcatcttctgg 126 DNA PAT source 1..126 mol_type other DNA organism synthetic construct misc_feature 1..126 note CD28 co-stimulatory domain gtgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccgggtccgacccgtaag cactaccaaccgtatgcgccgccgcgtgactttgcggcgtaccgtagc 336 DNA PAT source 1..336 mol_type other DNA organism synthetic construct misc_feature 1..336 note CD3 zeta primary signaling domain cgtgttaaatttagccgtagcgcggatgcgccggcgtaccagcagggtcagaaccaactgtataacgagctgaacct gggccgtcgtgaggaatatgacgtgctggataagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgta aaaacccgcaggaaggtctgtacaacgaactgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatga agggcgagcgtcgtcgtggtaaaggccacgatggtctgtaccagggcctgagcaccgcgaccaaagacacctatga tgcgctgcacatgcaagcgctgccgccgcgt 57 DNA PAT source 1..57 mol_type other DNA organism synthetic construct misc_feature 1..57 note viral self-cleaving 2A polypeptide gcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgggcccg 69 DNA

PAT source 1..69 mol_type other DNA organism synthetic construct misc_feature 1..69 note Signal peptide HLA-DQA1*05:01 atgatcctgaacaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggc 78 DNA

PAT source 1..78 mol_type other DNA organism synthetic construct misc_feature 1..78 note CD28 TM ttttgggttctggttgtggttggtggcgtgttagcttgctatagcctgctggttaccgtggcgtttattatcttctgg 126

DNA PAT source 1..126 mol_type other DNA organism synthetic construct misc_feature 1..126 note CD28 co-stimulatory domain gttcgcagcaagcgtagccgtctgctgcatagcgattacatgaatatgaccccgcgtcgtcctggcccgacccgcaaa cattatcaaccgtacgcgccgccgcgtgactttgcagcgtatcgtagc 336 DNA PAT source 1..336 mol_type other DNA organism synthetic construct misc_feature 1..336 note CD3 zeta primary signaling domain cgtgttaagtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaactgaatctg ggtcgtcgtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaaccgcgtcgtaa gaacccgcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatcggtatgaag ggcgaacgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggatacctacgacg cgctgcatatgcaggcgctgccgccgcgt 591 DNA PAT source 1..591 mol_type other DNA organism synthetic construct misc_feature 1..591 note HLA-DQA1*05:01 gaggacattgtggcggatcacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacac ccacgagttcgacggtgatgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgt cagttccgttttgatccgcaattcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgta gcaacagcaccgcggcgaccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagcc gaacatcctgatttgcctggttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtga ccgagggcgttagcgaaaccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgcc gagcgcggaggaaagctatgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagcc ggaaattccggcgccgatgagcgagctgaccgaaaccgttgtgtgc 866 AA PAT source 1..866 mol_type protein organism synthetic construct REGION 1..866 note CHAR _HLA-

DQ2.5-glia-omega1-glia-omega2 molecule (omega-gliadin)

MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGPQQPFPQPEQPFPWQPQGSGSGSLGSGSG

SGSGSRDSPEDFVYQFKGMCYFTNGTERVRLVSRSIYNREEIVRFDSDVGEFRAVTLLGLPAA

EYWNSQKDILERKRAAVDRVCRHNYQLELRTTLQRRVEPTVTISPSRTEALNHHNLLVCSVTD

FYPAQIKVRWFRNDQEETAGVVSTPLIRNGDWTFQILVMLEMTPQRGDVYTCHVEHPSLQSPI

TVEWRAQSESAQSKFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPG

PTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRD

PEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDA

LHMQALPPRGSGATNFSLLKQAGDVEENPGPMILNKALMLGALALTTVMSPCGGEDIVADHV

ASYGVNLYQSYGPSGQYTHEFDGDEQFYVDLGRKETVWCLPVLRQFRFDPQFALTNIAVLKH

NLNSLIKRSNSTAATNEVPEVTVFSKSPVTLGQPNILICLVDNIFPPVVNITWLSNGHSVTEGV

SETSFLSKSDHSFFKISYLTLLPSAEESYDCKVEHWGLDKPLLKHWEPEIPAPMSELTETVVCF

WVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA

AYRSRVKFSRSADAPAYQQGONQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEG

LYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR 9 AA

PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note HLA-

DQ2-glia-omegal PFPQPEQPF 9 AA PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note HLA-DQ2-glia-omega2 PQPEQPFPW 11 AA PAT source 1..11 mol_type protein organism unidentified REGION 1..11 note Complete gliadin omega peptide (HLA-DQ2-glia-omegal and HLA-DQ2-glia-omega2)

PFPQPEQPFPW 2601 DNA PAT source 1..2601 mol_type other DNA organism synthetic construct misc_feature 1..2601 note CHAR_HLA-DQ2.5-glia-omega1-glia- omega2 molecule (omega-gliadin) atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggtccacaacaaccttttccacagccggaacaaccatttccctggcaaccacaaggtagcg gtagcggtagcctgggcagcggcagcggtagcggtagcggcagccgtgacagcccggaggatttcgtttaccaattc aagggcatgtgctacttcaccaacggcaccgaacgtgtgcgtctggttagccgtagcatctacaaccgtgaggaaatt gtgcgtttcgacagcgatgttggcgagtttcgtgcggtgaccctgctgggtctgccggcggcggagtactggaacagc cagaaggacatcctggaacgtaaacgtgcggcggtggatcgtgtttgccgtcacaactatcagctggagctgcgtacc accctgcaacgtcgtgtggaaccgaccgttaccatcagcccgagccgtaccgaagcgctgaaccaccacaacctgct ggtgtgcagcgttaccgacttctacccggcgcagattaaagttcgttggtttcgtaacgatcaagaggaaaccgcggg tgtggttagcaccccgctgatccgtaacggcgactggaccttccagattctggttatgctggagatgaccccgcaacgt ggtgatgtgtacacctgccacgttgaacacccgagcctgcagagcccgattaccgtggagtggcgtgcgcagagcga aagcgcgcaaagcaagttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgc gttcatcatcttctgggtgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccg ggtccgacccgtaagcactaccaaccgtatgcgccgcegcgtgactttgcggcgtaccgtagccgtgttaaatttagcc gtagcgcggatgcgccggcgtaccagcagggtcagaaccaactgtataacgagctgaacctgggccgtcgtgagga atatgacgtgctggataagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgtaaaaacccgcaggaa ggtctgtacaacgaactgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatgaagggcgagcgtcgtc gtggtaaaggccacgatggtctgtaccagggcctgagcaccgcgaccaaagacacctatgatgcgctgcacatgcaa gcgctgccgccgcgtggtagcggtgcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgg gcccgatgatcctgaacaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggcgag gacattgtggcggatcacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacaccca cgagttcgacggtgatgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgtcag ttccgttttgatccgcaattcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgtagca acagcaccgcggcgaccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagccgaa catcctgatttgcctggttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtgaccg agggcgttagcgaaaccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgccga gcgcggaggaaagctatgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagccgg aaattccggcgccgatgagcgagctgaccgaaaccgttgtgtgcttttgggttctggttgtggttggtggcgtgttagct tgctatagcctgctggttaccgtggcgtttattatcttctgggttcgcagcaagcgtagccgtctgctgcatagcgattac atgaatatgaccccgcgtcgtcctggcccgacccgcaaacattatcaaccgtacgcgccgccgcgtgactttgcagcgt atcgtagccgtgttaagtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaa ctgaatctgggtcgtcgtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaacc gcgtcgtaagaacccgcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatc ggtatgaagggcgaacgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggata cctacgacgcgctgcatatgcaggcgctgccgccgcgttaa 27 DNA PAT source 1..27 mol_type unassigned DNA organism unidentified misc_feature 1..27 note HLA-DQ2-glia- omegal ccttttccacagccggaacaaccattt 27 DNA PAT source 1..27 mol_type unassigned DNA organism unidentified misc_feature 1..27 note HLA-DQ2-glia- omega2 ccacagccggaacaaccatttccctgg 33 DNA PAT source 1..33 mol_type unassigned DNA organism unidentified misc_feature 1..33 note Complete gliadin peptide (HLA-DQ2-glia-omegal and HLA-DQ2-glia-omega2) ccttttccacagccggaacaaccatttccctgg 2601 DNA PAT source 1..2601 mol_type other

DNA organism synthetic construct misc_feature 1..2601 note CHAR_HLA-DQ2.5- glia-omegal-glia-omega2 molecule (omega-gliadin) atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggtccacagcagcccttccctcagccagagcagccctttccttggcagccccagggtagcg gtagcggtagcctgggcagcggcagcggtagcggtagcggcagccgtgacagcccggaggatttcgtttaccaattc aagggcatgtgctacttcaccaacggcaccgaacgtgtgcgtctggttagccgtagcatctacaaccgtgaggaaatt gtgcgtttcgacagcgatgttggcgagtttcgtgcggtgaccctgctgggtctgccggcggcggagtactggaacagc cagaaggacatcctggaacgtaaacgtgcggcggtggatcgtgtttgccgtcacaactatcagctggagctgcgtacc accctgcaacgtcgtgtggaaccgaccgttaccatcagcccgagccgtaccgaagcgctgaaccaccacaacctgct ggtgtgcagcgttaccgacttctacccggcgcagattaaagttcgttggtttcgtaacgatcaagaggaaaccgcggg tgtggttagcaccccgctgatccgtaacggcgactggaccttccagattctggttatgctggagatgaccccgcaacgt ggtgatgtgtacacctgccacgttgaacacccgagcctgcagagcccgattaccgtggagtggcgtgcgcagagcga aagcgcgcaaagcaagttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgc gttcatcatcttctgggtgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccg ggtccgacccgtaagcactaccaaccgtatgcgccgccgcgtgactttgcggcgtaccgtagccgtgttaaatttagcc gtagcgcggatgcgccggcgtaccagcagggtcagaaccaactgtataacgagctgaacctgggccgtcgtgagga atatgacgtgctggataagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgtaaaaacccgcaggaa ggtctgtacaacgaactgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatgaagggcgagcgtcgtc gtggtaaaggccacgatggtctgtaccagggcctgagcaccgcgaccaaagacacctatgatgcgctgcacatgcaa gcgctgccgccgcgtggtagcggtgcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgg gcccgatgatcctgaacaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggcgag gacattgtggcggatcacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacaccca cgagttcgacggtgatgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgtcag ttccgttttgatccgcaattcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgtagca acagcaccgcggcgaccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagccgaa catcctgatttgcctggttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtgaccg agggcgttagcgaaaccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgccga gcgcggaggaaagctatgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagccgg aaattccggcgccgatgagcgagctgaccgaaaccgttgtgtgcttttgggttctggttgtggttggtggcgtgttagct tgctatagcctgctggttaccgtggcgtttattatcttctgggttcgcagcaagcgtagccgtctgctgcatagcgattac atgaatatgaccccgcgtcgtcctggcccgacccgcaaacattatcaaccgtacgcgccgccgcgtgactttgcagcgt atcgtagccgtgttaagtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaa ctgaatctgggtcgtcgtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaacc gcgtcgtaagaacccgcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatc ggtatgaagggcgaacgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggata cctacgacgcgctgcatatgcaggcgctgccgccgcgttaa 27 DNA PAT source 1..27 mol_type other DNA organism synthetic construct misc_feature 1..27 note HLA-DQ2-glia- omegal cccttccctcagccagagcagcccttt 27 DNA PAT source 1..27 mol_type other DNA organism synthetic construct misc_feature 1..27 note HLA-DQ2-glia-omega2 cctcagccagagcagccctttccttgg 33 DNA PAT source 1..33 mol_type other DNA organism synthetic construct misc_feature 1..33 note Complete gliadin peptide (HLA-DQ2-glia-omegal and HLA-DQ2-glia-omega2) cccttccctcagccagagcagccctttccttgg 888 AA PAT source 1..888 mol_type protein organism synthetic construct REGION 1..888 note CHAR_HLA-DQ2.5-glia- omegalomega2-glia-alphalalpha2 molecule (omega-gliadin and alpha-gliadin)

MSWKKALRIPGGLRAATVTLMLSMLSTPVAEGPQQPFPQPEQPFPWQPQGGGGGQLQPFPQ

PELPYPQPQLGSGSGSLGSGSGSGSGSRDSPEDFVYQFKGMCYFTNGTERVRLVSRSIYNRE

EIVRFDSDVGEFRAVTLLGLPAAEYWNSQKDILERKRAAVDRVCRHNYQLELRTTLQRRVEPT

VTISPSRTEALNHHNLLVCSVTDFYPAQIKVRWFRNDQEETAGVVSTPLIRNGDWTFQILVML

EMTPQRGDVYTCHVEHPSLQSPITVEWRAQSESAQSKFWVLVVVGGVLACYSLLVTVAFIIFW

VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQL

YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERR

RGKGHDGLYQGLSTATKDTYDALHMQALPPRGSGATNFSLLKQAGDVEENPGPMILNKALML

GALALTTVMSPCGGEDIVADHVASYGVNLYQSYGPSGQYTHEFDGDEQFYVDLGRKETVWCL

PVLRQFRFDPQFALTNIAVLKHNLNSLIKRSNSTAATNEVPEVTVFSKSPVTLGQPNILICLVDN

IFPPVVNITWLSNGHSVTEGVSETSFLSKSDHSFFKISYLTLLPSAEESYDCKVEHWGLDKPLL

KHWEPEIPAPMSELTETVVCFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMT

PRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDK

RRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATK

DTYDALHMQALPPR 5 AA PAT source 1..5 mol_type protein organism synthetic construct REGION 1..5 note Linker 5xG GGGGG 2667 DNA PAT source 1..2667 mol_type other DNA organism synthetic construct misc_feature 1..2667 note

CHAR_HLA-DQ2.5-glia-omegalomega2-glia-alphalalpha2 molecule (omega-gliadin and alpha-gliadin) atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggtccacaacaaccttttccacagccggaacaaccatttccctggcaaccacaaggtggtg gtggtggtcaacttcaaccttttcctcaacctgaacttccttatcctcaacctcaacttggtagcggtagcggtagcctgg gcagcggcagcggtagcggtagcggcagccgtgacagcccggaggatttcgtttaccaattcaagggcatgtgctac ttcaccaacggcaccgaacgtgtgcgtctggttagccgtagcatctacaaccgtgaggaaattgtgcgtttcgacagc gatgttggcgagtttcgtgcggtgaccctgctgggtctgccggcggcggagtactggaacagccagaaggacatcct ggaacgtaaacgtgcggcggtggatcgtgtttgccgtcacaactatcagctggagctgcgtaccaccctgcaacgtcg tgtggaaccgaccgttaccatcagcccgagccgtaccgaagcgctgaaccaccacaacctgctggtgtgcagcgttac cgacttctacccggcgcagattaaagttcgttggtttcgtaacgatcaagaggaaaccgcgggtgtggttagcacccc gctgatccgtaacggcgactggaccttccagattctggttatgctggagatgaccccgcaacgtggtgatgtgtacacc tgccacgttgaacacccgagcctgcagagcccgattaccgtggagtggcgtgcgcagagcgaaagcgcgcaaagca agttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgcgttcatcatcttctggg tgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccgggtccgacccgtaagc actaccaaccgtatgcgccgccgcgtgactttgcggcgtaccgtagccgtgttaaatttagccgtagcgcggatgcgcc ggcgtaccagcagggtcagaaccaactgtataacgagctgaacctgggccgtcgtgaggaatatgacgtgctggat aagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgtaaaaacccgcaggaaggtctgtacaacgaac tgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatgaagggcgagcgtcgtcgtggtaaaggccacg atggtctgtaccagggcctgagcaccgcgaccaaagacacctatgatgcgctgcacatgcaagcgctgccgccgcgt ggtagcggtgcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgggcccgatgatcctga acaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggcgaggacattgtggcggat cacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacacccacgagttcgacggtga tgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgtcagttccgttttgatccgca attcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgtagcaacagcaccgcggcga ccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagccgaacatcctgatttgcctgg ttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtgaccgagggcgttagcgaaa ccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgccgagcgcggaggaaagct atgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagccggaaattccggcgccgat gagcgagctgaccgaaaccgttgtgtgcttttgggttctggttgtggttggtggcgtgttagcttgctatagcctgctggt taccgtggcgtttattatcttctgggttcgcagcaagcgtagccgtctgctgcatagcgattacatgaatatgaccccgc gtcgtcctggcccgacccgcaaacattatcaaccgtacgcgccgccgcgtgactttgcagcgtatcgtagccgtgttaa gtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaactgaatctgggtcgtc gtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaaccgcgtcgtaagaaccc gcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatcggtatgaagggcga acgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggatacctacgacgcgctgc atatgcaggcgctgccgccgcgttaa 15 DNA PAT source 1..15 mol_type other DNA organism synthetic construct misc_feature 1..15 note Linker 5xG ggcggcggcggcggc 2667 DNA PAT source 1..2667 mol_type other DNA organism synthetic construct misc_feature 1..2667 note CHAR_HLA-DQ2.5-glia-omegalomega2-glia- alphalalpha2 molecule (omega-gliadin and alpha-gliadin) atgagctggaagaaagcgctgcgtatcccgggtggcctgcgtgcggcgaccgttaccctgatgctgagcatgctgag caccccggtggcggagggtccacagcagcccttccctcagccagagcagccctttccttggcagccccagggcggcg gcggcggccagctgcaaccgtttccgcagccggaactgccgtacccgcagccgcaactgggtagcggtagcggtag cctgggcagcggcagcggtagcggtagcggcagccgtgacagcccggaggatttcgtttaccaattcaagggcatgt gctacttcaccaacggcaccgaacgtgtgcgtctggttagccgtagcatctacaaccgtgaggaaattgtgcgtttcga cagcgatgttggcgagtttcgtgcggtgaccctgctgggtctgccggcggcggagtactggaacagccagaaggaca tcctggaacgtaaacgtgcggcggtggatcgtgtttgccgtcacaactatcagctggagctgcgtaccaccctgcaac gtcgtgtggaaccgaccgttaccatcagcccgagccgtaccgaagcgctgaaccaccacaacctgctggtgtgcagc gttaccgacttctacccggcgcagattaaagttcgttggtttcgtaacgatcaagaggaaaccgcgggtgtggttagca ccccgctgatccgtaacggcgactggaccttccagattctggttatgctggagatgaccccgcaacgtggtgatgtgta cacctgccacgttgaacacccgagcctgcagagcccgattaccgtggagtggcgtgcgcagagcgaaagcgcgcaa agcaagttttgggttctggtggttgtgggtggcgtgctggcgtgctacagcctgctggtgaccgttgcgttcatcatcttc tgggtgcgtagcaaacgtagccgtctgctgcacagcgactatatgaacatgaccccgcgtcgtccgggtccgacccgt aagcactaccaaccgtatgcgccgccgcgtgactttgcggcgtaccgtagccgtgttaaatttagccgtagcgcggat gcgccggcgtaccagcagggtcagaaccaactgtataacgagctgaacctgggccgtcgtgaggaatatgacgtgc tggataagcgtcgtggtcgtgatccggaaatgggtggcaagccgcgtcgtaaaaacccgcaggaaggtctgtacaa cgaactgcaaaaggacaaaatggcggaggcgtatagcgaaattggtatgaagggcgagcgtcgtcgtggtaaagg ccacgatggtctgtaccagggcctgagcaccgcgaccaaagacacctatgatgcgctgcacatgcaagcgctgccgc cgcgtggtagcggtgcgaccaacttcagcctgctgaagcaggcgggtgacgttgaggaaaacccgggcccgatgat cctgaacaaagcgctgatgctgggtgcgctggcgctgaccaccgttatgagcccgtgcggtggcgaggacattgtgg cggatcacgttgcgagctacggcgtgaacctgtaccagagctatggtccgagcggccaatacacccacgagttcgac ggtgatgaacaattttatgttgacctgggccgtaaggaaaccgtgtggtgcctgccggttctgcgtcagttccgttttga tccgcaattcgcgctgaccaacatcgcggtgctgaagcacaacctgaacagcctgattaaacgtagcaacagcaccg cggcgaccaacgaggttccggaagtgaccgttttcagcaaaagcccggtgaccctgggtcagccgaacatcctgattt gcctggttgacaacatctttccgccggttgtgaacattacctggctgagcaacggtcacagcgtgaccgagggcgtta gcgaaaccagcttcctgagcaagagcgatcacagcttctttaaaatcagctatctgaccctgctgccgagcgcggagg aaagctatgactgcaaggtggagcactggggtctggataagccgctgctgaaacactgggagccggaaattccggc gccgatgagcgagctgaccgaaaccgttgtgtgcttttgggttctggttgtggttggtggcgtgttagcttgctatagcct gctggttaccgtggcgtttattatcttctgggttcgcagcaagcgtagccgtctgctgcatagcgattacatgaatatga ccccgcgtcgtcctggcccgacccgcaaacattatcaaccgtacgcgccgccgcgtgactttgcagcgtatcgtagccg tgttaagtttagccgtagcgcggacgcgccggcgtatcaacagggccaaaatcagctgtacaatgaactgaatctgg gtcgtcgtgaagagtacgatgttctggacaaacgtcgtggtcgtgacccggagatgggtggcaaaccgcgtcgtaag aacccgcaggaaggtttatataatgagctgcagaaagataagatggcggaagcgtatagcgaaatcggtatgaagg gcgaacgtcgtcgtggcaagggtcatgacggcctgtatcaaggtctgagcaccgcgaccaaggatacctacgacgc gctgcatatgcaggcgctgccgccgcgttaa 15 DNA PAT source 1..15 mol_type other DNA organism synthetic construct misc_feature 1..15 note Linker 5xG ggcggcggcggcggc 9 AA PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note

HLA-DQ8-glia-alphal EGSFQPSQE 27 DNA PAT source 1..27 mol_type unassigned

DNA organism unidentified misc_feature 1..27 note alternative CLIP peptide ccgctgctgatgcaggcgctgccgatg 14 AA PAT source 1..14 mol_type protein organism unidentified REGION 1..14 note Complete alpha gliadin peptide (HLA-DQ2-glia- alphala and HLA-DQ2-glia-alpha2) with 5' flanking QLQPFPQPELPYPQ 17 AA PAT source 1..17 mol_type protein organism unidentified REGION 1..17 note Complete alpha gliadin peptide (HLA-DQ2-glia-alphala and HLA-DQ2-glia-alpha2) with 5' flanking and 3' flanking QLQPFPQPELPYPQPQL 14 AA PAT source 1..14 mol_type protein organism unidentified REGION 1..14 note Complete alpha gliadin peptide (HLA-DQ2-glia-alphala and HLA-DQZ2-glia-alpha2) with 3' flanking

PFPQPELPYPQPQL 14 AA PAT source 1..14 mol_type protein organism unidentified

REGION 1..14 note Complete gliadin omega peptide (HLA-DQ2-glia-omegal and

HLA-DQ2-glia-omega2) with 5' flanking PQQPFPQPEQPFPW 17 AA PAT source 1..17 mol_type protein organism unidentified REGION 1..17 note Complete gliadin omega peptide (HLA-DQ2-glia-omegal and HLA-DQ2-glia-omega2) with 5' flanking and 3' flanking PQQPFPQPEQPFPWQPQ 14 AA PAT source 1..14 mol_type protein organism unidentified REGION 1..14 note Complete gliadin omega peptide (HLA-DQ2-glia- omegal and HLA-DQ2-glia-omega2) with 3' flanking PFPQPEQPFPWQPQ 9 AA PAT source 1..9 mol_type protein organism unidentified REGION 1..9 note HLA-DQ2- glia-alphalb PYPQPELPY 11 AA PAT source 1..11 mol_type protein organism unidentified REGION 1..11 note Complete gliadin alpha peptide (HLA-DQ2-glia- alphalb and HLA-DQZ2-glia-alpha2) PYPQPELPYPQ 14 AA PAT source 1..14 mol_type protein organism unidentified REGION 1..14 note Complete alpha gliadin peptide (HLA-DQ2-glia-alphalb and HLA-DQ2-glia-alpha2) with 5' flanking

QLQPYPQPELPYPQ 17 AA PAT source 1..17 mol_type protein organism unidentified

REGION 1..17 note Complete alpha gliadin peptide (HLA-DQ2-glia-alphalb and

HLA-DQ2-glia-alpha2) with 5' flanking and 3' flanking QLQPYPQPELPYPQPQL 14 AA

PAT source 1..14 mol_type protein organism unidentified REGION 1..14 note

Complete alpha gliadin peptide (HLA-DQ2-glia-alphalb and HLA-DQ2-glia-alpha2) with 3' flanking PYPQPELPYPQPQL

Claims

Conclusions

1. Chimeric HLA class II molecule, comprising:

a. a first polypeptide comprising:

1. a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 8162 chain, and u. a transmembrane domain; and b. a second polypeptide comprising:

i. a second extracellular domain comprising an HLA-DQ ala2 chain, and u. a transmembrane domain, wherein the first and/or second polypeptide further comprises (iii) an immune receptor intracellular signalling domain; wherein the HLA-DQ ala2 and δ162 chains are encoded by the HLA-DQA1*05 and HLA-DQB1*02 genes, respectively; or wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*02 and HLA-DGQGB1*02 genes, respectively, or wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*03 and HLA-DQB1*03 genes, respectively.

2. Fusion polypeptide comprising, at an N-terminus relative to C-terminal orientation:

a. a first extracellular domain comprising a gluten-derived peptide comprising one or more gluten-derived epitopes and an HLA-DQ 5162 chain;

b. a transmembrane domain;

c. a peptide cleavage signal;

d. a second extracellular domain comprising an HLA-DQ ala2 chain;

e. a transmembrane domain; wherein the polypeptide further comprises an immunoreceptor intracellular signalling domain located at the C-terminus of (b) and/or (e), and wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*05 and HLA-DQB1*02 genes, respectively, or wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*02 and HLA-DQB1*02 genes, respectively, or wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*03 and HLA-DQB1*03 genes, respectively.

The fusion polypeptide of claim 2, further comprising an HLA-DQ B182 chain signal sequence located at the N-terminus relative to the first extracellular domain; and an HLA-DQ ala2 chain signal sequence located at the N-terminus relative to the second extracellular domain.

The chimeric HLA class II molecule or fusion polypeptide of any preceding claim, wherein the one or more gluten-derived epitopes are derived from a protein selected from the group consisting of: gliadin, glutenin, hordein, secalin, and avenin.

The chimeric HLA class II molecule or fusion polypeptide of claim 4, wherein the gliadin is alpha-gliadin or omega-gliadin.

The chimeric HLA class II molecule or fusion polypeptide of claim 5, wherein the gliadin is alpha-gliadin.

The chimeric HLA class II molecule or fusion polypeptide of claim 6, wherein the one or more gluten-derived epitopes comprise the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 69.

The chimeric HLA class II molecule or fusion polypeptide of claim 6, wherein the one or more gluten-derived epitopes comprise the amino acid sequence of SEQ ID NO: 61.

9. Chimeric HLA class II molecule or fusion polypeptide according to any one of the preceding claims, wherein the gluten-derived peptide comprises two or more gluten-derived epitopes.

The chimeric HLA class II molecule or fusion polypeptide of claim 9, wherein the gluten-derived peptide comprises the amino acid sequences of SEQ ID NO: 3 and SEQ ID NO: 4, or SEQ ID NO: 69 and SEQ ID NO: 4.

The chimeric HLA class II molecule or fusion polypeptide of claim 1, wherein the gluten-derived peptide comprises the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 70.

The chimeric HLA class II molecule or fusion polypeptide of claim 5, wherein the gliadin is omega-gliadin.

The chimeric HLA class II molecule or fusion polypeptide of claim 12, wherein the one or more gluten-derived epitopes comprise the amino acid sequence of SEQ ID NO: 44.

The chimeric HLA class II molecule or fusion polypeptide of claim 12, wherein the one or more gluten-derived epitopes comprise the amino acid sequence of SEQ ID NO: 45.

The chimeric HLA class II molecule or fusion polypeptide of claim 9, wherein the gluten-derived peptide comprises the amino acid sequence of SEQ ID NO: 44 and SEQ ID NO: 45.

The chimeric HLA class II molecule or fusion polypeptide of claim 15, wherein the gluten-derived peptide comprises the amino acid sequence of SEQ ID NO: 46.

17. Chimeric HLA class II molecule or fusion polypeptide according to any one of the preceding claims, wherein the gluten-derived peptide comprises four or more gluten-derived epitopes.

The chimeric HLA class II molecule or fusion polypeptide of claim 17, wherein the gluten-derived peptides are selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 69, SEQ ID NO: 44, and SEQ ID NO: 45.

The chimeric HLA class II molecule or fusion polypeptide of claim 18, wherein the gluten-derived peptide comprises:

a. the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 46, optionally wherein the gluten-derived peptide comprises a linker between the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 46, or b. an amino acid sequence of SEQ ID NO: 70 and SEQ ID NO: 46, optionally wherein the gluten-derived peptide comprises a linker between the amino acid sequence of SEQ ID NO: 70 and SEQ ID NO: 46.

20. The chimeric HLA class II molecule or fusion polypeptide of any preceding claim, wherein the first extracellular domain further comprises a linker sequence between the gluten-derived peptide and the HAL-DQ B162 chain.

21. Chimeric HLA class II molecule or fusion polypeptide according to any one of the preceding claims, wherein the gluten-derived peptide further comprises adjacent amino acids at the N- and/or C-terminal side of the gluten-derived epitope, optionally wherein the adjacent amino acids are natural adjacent amino acids

22. Chimeric HLA class II molecule or fusion polypeptide according to any one of the preceding claims, wherein the transmembrane domain is selected from the group consisting of: alpha or beta chain of CD28, CD4, CD5, CDS,

CD9, CD16, CD22, CD27, CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137, CD152, CD154, PD1, HLA-DR, HLA-DQ or HLA-DP.

The chimeric HLA class II molecule or fusion polypeptide of claim 22, wherein the transmembrane domain is a CD28 transmembrane domain.

24. The chimeric HLA class II molecule or fusion polypeptide of any preceding claim, wherein the immune receptor intracellular signaling domain comprises one or more costimulatory signaling domains.

The chimeric HLA class II molecule or fusion polypeptide of claim 24, wherein the one or more costimulatory signaling domains are selected from the group consisting of: TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLRS, TLR9, TLR10, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (0X40), CD137 (4-1BB), CD154 (CD40L), CD278 (ICOS), DAP10, LAT, NKD2C, SLP76, TRIM, and ZAP70 costimulatory signaling domain.

The chimeric HLA class II molecule or fusion polypeptide of claim 25, wherein the one or more co-signaling domains is or are a CD28 co-stimulatory signaling domain.

27. The chimeric HLA class II molecule or fusion polypeptide of any preceding claim, wherein the immune receptor intracellular signaling domain comprises a primary signaling domain.

The chimeric HLA class II molecule or fusion polypeptide of claim 27, wherein the primary signaling domain is selected from the group consisting of: FcRγ, FcRB, CD3γ, CD3β, CD3ε, CD3β, CD22, CD79b, and CD66d.

29. The chimeric HLA class II molecule or fusion polypeptide of claim 28, wherein the primary signaling domain is CD3{.

30. Nucleic acid encoding a chimeric HLA class II molecule or fusion polypeptide, according to any one of the preceding claims, optionally wherein the nucleic acid is RNA and/or DNA.

31. Vector comprising a nucleic acid according to claim 30.

32. Cell comprising one or more nucleic acids, vectors, chimeric HLA class II molecules and/or fusion polypeptides according to any of the preceding claims.

The cell of claim 32, wherein the cell is a T cell, optionally wherein the T cell is selected from the group consisting of a CD8+ T cell, an NK T cell, a CD3+ T cell, and a v0 T cell, or a mixture of any of the foregoing.

The cell of claim 33, wherein the cell is an NK cell or a natural lymphoid cell (ILC).

35. Cell, comprising at least two of the following:

a. a first nucleic acid, vector, chimeric HLA class II molecule, or fusion polypeptide according to any preceding claim, wherein the HLA-DQ ala2 and B1B2 chains are encoded by the HLA-DQAI*05 and HLA-DQBI1*02 genes, respectively,

b. a second nucleic acid, vector, chimeric HLA class II molecule, or fusion polypeptide according to any one of the preceding claims, wherein the HLA-DQ ala2 and δ182 chains are encoded by the HLA-DQA1*02 and HLA-DQB1*02 genes, respectively; and/or c. a third nucleic acid, vector, chimeric HLA class II molecule, or fusion polypeptide according to any one of the preceding claims, wherein the HLA-DQ ala2 and δ162 chains are encoded by the HLA-DQA1*03 and HLA-DQB1*03 genes, respectively.

The cell of claim 35, wherein the HLA-DQA1*02 and HLA-DQB1*02 are HLA-DQA1*02:01 and HLA-DQB1*02:02, respectively.

The cell of claim 35 or 36, wherein the HLA-DQA1*05 and HLA-DQBI*02 are HLA-DQA1*05:01 and HLA-DQB1*02:01, respectively.

38. A composition comprising a nucleic acid, a vector, a chimeric HLA class II molecule, a fusion polypeptide and/or a cell according to any one of the preceding claims.

39. A pharmaceutical composition comprising a nucleic acid, a vector, a chimeric HLA class II molecule, a fusion polypeptide, and/or a cell according to any preceding claim.

40. A pharmaceutical composition according to claim 39 for use as a medicament.

41. A pharmaceutical composition according to claim 39 for use in treating or preventing celiac disease in an HLA-DQ positive subject.

42. The pharmaceutical composition for use according to claim 41, wherein the subject is HLA-DQA1*02-HLA-DQB1*02, HLA-DQA1*05-HLA-DQB1*02, and/or HLA-DQA1*03-DQB1*03.