[go: up one dir, main page]

WO2017177175A1 - Procédés et compositions ciblant une latence rétrovirale - Google Patents

Procédés et compositions ciblant une latence rétrovirale Download PDF

Info

Publication number
WO2017177175A1
WO2017177175A1 PCT/US2017/026669 US2017026669W WO2017177175A1 WO 2017177175 A1 WO2017177175 A1 WO 2017177175A1 US 2017026669 W US2017026669 W US 2017026669W WO 2017177175 A1 WO2017177175 A1 WO 2017177175A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
cell
antibody
ifitml
hiv
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2017/026669
Other languages
English (en)
Inventor
Rui Andre Saraiva RAPOSO
Miguel de Mulder ROUGVIE
Dominic Paquin PROULX
Phillip BRAILEY
Vincius D. CABIDO
Paul Zdinak
Richard B. Jones
Douglas F. Nixon
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
George Washington University
Original Assignee
George Washington University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by George Washington University filed Critical George Washington University
Publication of WO2017177175A1 publication Critical patent/WO2017177175A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/46—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C12N5/0638—Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells

Definitions

  • This invention relates generally to compositions and methods for reducing and eradicating reservoirs of latent viruses.
  • ART Antiretroviral therapy
  • HAART highly-active antiretroviral therapy
  • Latent viral reservoirs are thought to be maintained by their intrinsic stability and homeostatic proliferation of latent cells, reseeding of newly infected cells by low levels of replicating HIV-1 in lymph nodes and gut-associated lymphoid tissue (GALT) and incomplete penetration of antiretroviral drugs into infected tissues. HIV-1 may continue to replicate and traffic in lymphoid tissue despite potent ART. Viral DNA persists in central memory and stem central memory CD4+ T-cells.
  • Current eradication strategies rely on histone deacetylase inhibitors (HDACi) and related compounds, in conjugation with ART, to reactivate and kill infected cells within the reservoir.
  • HDACi histone deacetylase inhibitors
  • the invention provides novel therapeutic methods and compositions directed to reactivating latent viral reservoirs and eliminating them.
  • one or more biomarkers are provided, including IFITM1, that identify the quantity of HIV latently infected cells during treatment and thus quantify the latent reservoir as a monitoring method that assesses the efficacy of treatment and directs modifications that are able to attain reduced expression of IFITM1.
  • a method of treatment that targets IFITM1+ HIV infected cells by the use of an antibody that binds to IFITM1.
  • the antibody that binds to IFITMl is not cross-reactive with IFITM2 and IFITM3.
  • the antibody is bi-specific.
  • the bi-specific antibody includes an anti-IFITMl moiety and an anti-CD4 moiety.
  • the antibody is co-administered with an effective amount of an NK cell stimulator.
  • the NK cell stimulator is selected from one or more of type I IFNs (mainly IFN-a and IFN- ⁇ ), IL-2/anti-IL-2 monoclonal antibody complexes, IL-12, IL-15 and IL-18.
  • the antibody to IFITM1 and the NK cell stimulator synergistically kill cells latently infected with the latent immunodeficiency virus.
  • the NK cell stimulator is an IL-15 superagonist.
  • An example of an IL-15 superagonist is an IL-15 mutant bound to an IL-15 receptor a/IgGl Fc fusion protein.
  • the antibody including specificity for IFITM1 is a chimeric molecule and is conjugated to a therapeutic agent.
  • the therapeutic agent may be a cytotoxic agent selected from the group consisting of paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, a glucocorticoid, procaine, tetracaine, lidocaine, propranolol, puromycin, epirubicin, and cyclophosphamide.
  • a treatment regimen for reducing a pool of latently infected cells in a patient infected with a latent immunodeficiency virus comprising co-administering an anti-IFITMl specific monoclonal antibody and one or more anti-viral agents selected from viral entry inhibitors, nucleotide/side reverse transcription inhibitors (NRTI), non-nucleoside reverses transcriptase inhibitors (NNRTI), integrase inhibitors and protease inhibitors.
  • NRTI nucleotide/side reverse transcription inhibitors
  • NRTI non-nucleoside reverses transcriptase inhibitors
  • integrase inhibitors and protease inhibitors selected from viral entry inhibitors, nucleotide/side reverse transcription inhibitors (NRTI), non-nucleoside reverses transcriptase inhibitors (NNRTI), integrase inhibitors and protease inhibitors.
  • the treatment regimen may further comprise co-administering a drug that targets an immune checkpoint (IC) molecule, wherein the IC molecule is selected from Programed Cell Death- 1 (PD-1), Programed Cell Death Ligand-1 (PD-L1), T-cell Immunoregulator with Ig and ITEVI domains (TIGIT), Lymphocyte Activation Gene 3 (LAG-3), and combinations thereof.
  • IC immune checkpoint
  • the treatment regimen may include co-administering and effective amount of an NK cell stimulator selected from one or more of type I IFNs (mainly IFN-a and IFN- ⁇ ), IL- 2/anti-IL-2 monoclonal antibody complexes, IL-12, IL-15 and IL-18, and wherein the antibody to IFITM1 and the NK cell stimulator synergistically kill cells latently infected with the latent immunodeficiency virus.
  • an NK cell stimulator selected from one or more of type I IFNs (mainly IFN-a and IFN- ⁇ ), IL- 2/anti-IL-2 monoclonal antibody complexes, IL-12, IL-15 and IL-18, and wherein the antibody to IFITM1 and the NK cell stimulator synergistically kill cells latently infected with the latent immunodeficiency virus.
  • a cocktail of drugs is provided that is directed to killing cells latently infected with HIV wherein the cocktail comprises anti-IFITMl specific monoclonal antibody and a monoclonal antibody directed to an IC molecule selected from one or more of Programed Cell Death- 1 (PD-1), Programed Cell Death Ligand-1 (PD-L1), T-cell Immunoregulator with Ig and ITIM domains (TIGIT), and Lymphocyte Activation Gene 3 (LAG-3).
  • PD-1 Programed Cell Death- 1
  • PD-L1 Programed Cell Death Ligand-1
  • TAGIT T-cell Immunoregulator with Ig and ITIM domains
  • LAG-3 Lymphocyte Activation Gene 3
  • a CAR T cell is provided that has specificity for IFITM1.
  • a CAR T cell is bi-specific and includes an IFITM1 specific scFv joined in tandem to a CD4 specific scFv in the CAR extracellular domain. Whether mono or bi-specific, the exodomain is linked through a transmembrane domain to a signaling endodomain including one or more of signaling and co-stimulatory domains derived from CD2, CD 3, CD 27, CD28, CD30 (aka TNFRSF8), CD40 (aka TNFRSF5), CD!
  • the IFITM1 specific CAR T cells are expanded ex vivo using IL-15 prior to administering the CAR modified T cells to the immunodeficiency virus-infected host.
  • the mono or bi-specific CAR T cell that includes IFITM1 specificity is administered to HIV infected patients to reduce the load of latently infected cells.
  • the patient is co-administered a drug that targets an immune checkpoint (IC) molecule selected from Programed Cell Death- 1 (PD-1), Programed Cell Death Ligand-1 (PD-L1), T- cell Immunoregulator with Ig and ITIM domains (TIGIT), Lymphocyte Activation Gene 3 (LAG-3), and combinations thereof.
  • IC immune checkpoint
  • the patient may be co-administered one or more antiviral agents selected from viral entry inhibitors, nucleotide/side reverse transcription inhibitors ( RTI), non-nucleoside reverses transcriptase inhibitors (N RTI), integrase inhibitors and protease inhibitors.
  • RTI nucleotide/side reverse transcription inhibitors
  • N RTI non-nucleoside reverses transcriptase inhibitors
  • integrase inhibitors integrase inhibitors and protease inhibitors.
  • a method for treatment of a human individual infected with Human Immunodeficiency Virus comprising the steps of: (a) obtaining a sample of peripheral blood from the individual; (b) determining an expression level of IFITMl on T cells in the sample; (c) administering a treatment regimen to the individual; (d) periodically repeating steps (a) and (b) to determine efficacy of the administered treatment; and (e) adjusting the treatment regimen if needed to attain a reducing in IFITMl expression of T cells from the individual.
  • the treatment regimen may include administering one or more of: a cocktail of antiviral drugs, anti-IFITMl antibodies, IC antagonists, anti-IFITMl CAR T cells, and cytokines directed to maximizing NK function or CAR T cell activity.
  • Fig. 1A illustrates a scheme for generating latently infected CD4+ T-cells according to one model.
  • CD4+ T-cells were negatively selected from PBMC and conditioned for 3 days with CCL19.
  • CCL19-treated cells were infected with HIV-1NL4-3 for 6 days and resting CD4+ T-cells (CD25-/CD69-/HLA-DR-) were negatively selected by magnetic cell sorting. Resting CD4+ T-cells were reactivated with anti-CD3/CD28 for three days.
  • FIG. 2A shows quantification of HIV-1 gag by real-time qPCR.
  • Fig. 2B shows quantification of HIV-1 mRNA transcripts by real-time qPCR.
  • HIV negative cells are also stained for SSC-A.
  • Fig. 3B the HIV negative cells are co-stained with anti-CD3/CD28.
  • Fig. 3C resting latent cells are also stained for SSC-A.
  • Fig. 3D the resting latent cells are co- stained with anti-CD3/CD28.
  • Figs. 4A - 4F represent ADCC of latently infected ACH-2.
  • ACH-2 (target) cells were incubated with anti-IFITMl or an isotype control prior to addition of effector cells.
  • Target cell apoptosis was detected using the FLICA assay and a live/dead marker.
  • FIG. 4C depicts the overall percentage of killed (dead/FLICA+) ACH-2 target cells.
  • Fig. 4D depicts IFN- g+CD107a+NK cells.
  • Fig. 4E depicts IFN-g+NK cells and
  • Data are plotted as mean ⁇ SD and significance was determined using a non- parametric two-tailed t-test. P ⁇ 0.05 was considered significant.
  • Figs. 5 A - 5C show ADCC of IFITM1+CD4+ T-cells from ART-suppressed patients.
  • Naive, Stem central memory (TSCM), Central memory (TCM), Transitional memory (TTM) and Effector memory (TEM) cells are shown.
  • Fig. 5B depicts purified CD4+ T-cells from ART-suppressed patients incubated with anti-IFITMl or isotype control prior to the addition of autologous effector cells at an E:T ratio of 10: 1.
  • FIG. 5C depicts the results when cells were washed and stained with anti-CD3, CD56, CD107a and IFN-g to detect NK cell degranulation and production of intracellular IFN-g.
  • FIGs. 6A - 6G depict the results of reactivation of HIV-1 in ACH-2 cells.
  • ACH-2 cells were treated for 5h with increasing concentrations of SAHA, panobinostat and romidepsin. After the initial pulse, cells were washed twice in complete media and cultured for an additional 18h. The frequency of reactivated cells was determined by intracellular HIV-1 Gag staining. Representative flow plots of three independent experiments are shown.
  • FIG. 6E summarizing the results of Figs. 6B - 6D.
  • Reactivated ACH-2 cells were calculated by subtracting the percentage of HIV-1 Gag+ cells by the percentage of HIV- 1 Gag+ cells in untreated control. Data are plotted as mean ⁇ SEM.
  • Fig. 6G shows the effect of different HDACi on ACH-2 cell viability. Representative flow plots of three independent experiments are shown.
  • Figs. 7A - 7H show gating strategy for the detection of IFITM1 in different CD4+ T cell subsets.
  • Fig. 7A shows the side scatter area (SSC-A) plotted against the Forward Scatter area (FSC-A) by flow cytometry with the cells to be analyzed within the drawn area.
  • Fig. 7B shows the Forward Scatter area (FSC-A) plotted against the Forward Scatter height (FSC-H).
  • PBMC were stained with antibodies against CD3 (Fig. 7C), CD4 (Fig. 7D and F), CD 8 (Fig. 7D), CD45RA (Fig. 7F), CCR7 (Figs. 7E and G), CD27 (Figs. 7E and G), CD95 (Fig. 7H) and IFITMl .
  • Dead cells were excluded using a live/dead marker.
  • HIV-1 persistence in latent reservoirs during antiretroviral therapy is the main obstacle to virus eradication.
  • ART antiretroviral therapy
  • Latency models have predominantly been used as means to study HIV-1 reactivation in hopes of providing data towards "shock-and-kill" strategies.
  • IFITMl Interferon Induced Transmembrane Protein 1
  • IFITMl is used as a marker for natural HIV-1 reservoirs and provides an immune target for killing of latently infected cells through IFITMl -antibody-dependent cell-mediated cytolysis (ADCC) alone and combination with other agents.
  • IFITMl is used as a bio-marker for detecting and quantitating latently infected T cells.
  • antiviral regimens are tailored to maximize the efficacy of treatment in individual patients.
  • monitoring of the efficacy of an anti-HIV regimen in a patient comprises: determining levels of expression of IFITMl on T cells from the patient, treating the patient with a treatment regimen, periodically re-determining levels of expression of IFITMl on T cells from the patient, and adjusting the therapeutic regimen to maximally affect reduced levels of IFITMl expression.
  • the therapeutic regimen may include administering one or more of: a cocktail of antiviral drugs, anti-IFITMl antibodies, IC antagonists, anti-IFITMl CAR T cells, and cytokines directed to maximizing NK function or CAR T cell activity.
  • ELC ELC
  • MTP-3-beta MTP-3-beta
  • SAHA suberanilohydroxamic acid, a.k.a vorinostat is a histone deacetylase inhibitor marketed by Merck as ZOLINZA ®
  • SAMHD1 SAM domain and HD domain-containing protein 1, cellular enzyme that blocks HIV replication by converting dNTPs to inorganic phosphate and 2'-deoxynucleoside thus depelted available pool for RT
  • the phrase "at least one of when combined with a list of items, means a single item from the list or any combination of items in the list.
  • the phrase "at least one of A, B and C,” means “at least one from the group A, B, C, or any combination of A, B and C.”
  • the phrase requires one or more, and not necessarily not all, of the listed items.
  • immunodeficiency virus includes human immunodeficiency virus (HIV) as well as feline immunodeficiency virus and simian immunodeficiency virus.
  • HIV as used herein includes human immunodeficiency virus- 1 (HIV-1), human immunodeficiency virus-2 (HIV-2), and other HIV subtypes.
  • antibody includes both intact immunoglobulin molecules as well as portions, fragments, and derivatives thereof, such as, for example, Fab, Fab', F(ab')2, Fv, Fsc, CDR regions, or any portion of an antibody that is capable of binding an antigen or epitope including chimeric antibodies that are bi-specific or that combine an antigen binding domain originating with an antibody with another type of polypeptide.
  • a bi-specific antibody is utilized that includes antigen specific binding for IFITMl on one Fab domain and antigen specific binding for CD4 on the other Fab domain.
  • antibody includes monoclonal antibodies (mAb), chimeric antibodies, humanized antibodies, as well as fragments, portions, regions, or derivatives thereof, provided by any known technique including but not limited to, enzymatic cleavage and recombinant techniques.
  • antibody as used herein also includes single-domain antibodies (sdAb) and fragments thereof that have a single monomelic variable antibody domain (VH) of a heavy-chain antibody.
  • sdAb which lack variable light (VL) and constant light (CL) chain domains are natively found in camelids (VHH) and cartilaginous fish (VNAR) and are sometimes referred to as "Nanobodies” by the pharmaceutical company Ablynx who originally developed specific antigen binding sdAb in llamas.
  • VHH camelids
  • VNAR cartilaginous fish
  • the modifier "monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • co-administered and “in combination with” include administration with two or more therapeutic agents either simultaneously, concurrently or sequentially in any order without specific time limits so long as the two or more "co-administered” drugs are present in measureable amounts in a single patient at a given time.
  • the therapeutic agents are in the same composition or unit dosage form while in other embodiments, the therapeutic agents are in separate compositions or unit dosage forms.
  • active agent and synonymous terms according to those of skill in the art are used interchangeably herein.
  • An effective amount is an amount that reduces the reservoir of latent virus in an individual by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%.
  • Reductions in the reservoir of latently infected cells is can be determined using any known method, including by polymerase chain reaction (PCR), detecting and/or measuring viral polypeptides and antigens and measuring the CD4 + T cell count in the individual.
  • PCR polymerase chain reaction
  • an eradication strategy is provided selectively targeting IFITM1 in conjugation with targeting immune checkpoint molecules shown to contribute to HIV persistence during ART to the end of effectively targeting and eliminate latently infected cells in vivo.
  • Immune checkpoint molecules act to up or down-modulate immune responses.
  • Down-regulating (inhibitory) immune checkpoint molecules act normally to prevent hyper-immune activation, minimize collateral damage, and maintain peripheral self- tolerance.
  • ICs are normally up-regulated with T-cell activation and act to limit the effector response by feedback inhibition.
  • IC overexpression is associated with T-cell exhaustion and dysfunction in cancer and chronic viral infections, including HIV. Examples of immune check point molecules that may be targeted include those identified as expressed on cells harboring latent virus.
  • PD-1 Programed Cell Death- 1
  • PD-L1 Programed Cell Death Ligand-1
  • TAGIT T-cell Immunoregulator with Ig and ITEVI domains
  • LAG-3 Lymphocyte Activation Gene 3
  • PD-1 is a cell surface receptor that is a member of the CD28 family of T-cell regulators.
  • the human PD-1 gene is on chromosome 2q37 and the full-length PD-1 cDNA encodes a 288 amino acid protein.
  • Several monoclonal antibodies targeting PD-1 have been developed to reverse PD-1 mediated T-cell down-regulation of antitumor responses.
  • Anti PD-1 monoclonal developed to date include Nivolumab (previously termed BMS-936558, now FDA approved for certain cancers and marketed by Bristol-Myers Squibb as OPDIVO 3 ⁇ 4 ), Pembrolizumab (previously termed MK-3475 now FDA approved for certain cancers and marketed by Merck as EYTRUDA*), and Pidilizumab (previously termed CT- 01 1 by CureTech and MDV9300 by Mediation).
  • PD-1 has two ligands, PD-L1 and PD-L2, which are also transmembrane cell surface molecules that are typically expressed on antigen presenting cells and serve as immune system inhibitors. Expression of these ligands on tumor cells help the tumor ceils avoid immune mediated control.
  • Programed Death Ligand-1 (PD-L1) is also known as cluster of differentiation (CD) 274 or B7 homolog 1 is encoded in humans by the CD274 gene.
  • the IC molecule PD-L! may be targeted by Atezolizumab (tradename TECENTRTG® by Genentech) and Avelumab (currently co-developed by Pfizer and Merck and also known as MSB0G1Q718C).
  • LAG-3, CD designated as C ' 0223. is an immunoglobulin superfamiiy 503 -ami no acid transmembrane protein encoded on human chromosome 12 i 3 adjacent to the gene encoding CD4.
  • BMS- 986016 for tumor treatment.
  • GlaxoSmith iine has also developed an anti -LAG-3 monoclonal antibody (GSK283178I) but for the indication of treating psoriasis.
  • TIGIT is a member of CD28 family and includes an extracellular IgV domain, a transmembrane domain, and a cytoplasmic tail containing two-immimoreceptor tyrosine- based inhibitor ⁇ ' motifs (ITIM).
  • ITIM two-immimoreceptor tyrosine- based inhibitor ⁇ ' motifs
  • TIGIT and PD-1 have been shown to be overexpressed on viral iy exhausted T cells. See Chew, GM, et al. "TIGIT Marks Exhausted T Cells, Correlates with Disease Progression, and Serves as a Target for Immune Restoration in HIV and SIV infection" PLoS Pathog 12(1) (2016) 1-28.
  • Antibodies to TIGIT for immunomodulation are under development by Genentech for reversal of T-cell exhaustion in cancer (MTIG7192A, RG6058)
  • the IFITM 1 gene encodes the Leu-13 antigen, designated as CD225. As their name suggests, IFITM are strongly upregulated by both type I and II interferons.
  • IFITM family of proteins act to uniquely inhibit replication of members of the following diverse group of membrane-enveloped viruses: retroviruses (HIV), filoviruses (Marburg and Ebola), coronavirus (Severe Acute Respiratory Syndrome - SARS-CoV), phlebovirus (Rift Valley Fever - RVF), paramyxovirus (respiratory syncytial virus - RSV), flaviviruses (including Dengue (DenV), West Nile Virus (WNV), Zika, Yellow Fever Virus, and several hemorrhagic viruses)), influenza A virus and Hepatitis type C (HCV).
  • retroviruses HIV
  • filoviruses Marburg and Ebola
  • coronavirus severe Acute Respiratory Syndrome - SARS-CoV
  • phlebovirus Rospiratory syncytial virus - RSV
  • flaviviruses including Dengue (DenV), West Nile Virus (WNV), Zi
  • restriction factors possess potent anti- HIV-1 activities by interfering with the virus life cycle. Restriction factor expression varies between cells and activation states, and in addition to acting directly against HIV-1, they elicit immune responses to accelerate the clearance of infected cells. Such roles have been described for BST2/tetherin and TRIM5. Type-I interferons, such as IFNa, are able to strongly induce the expression of most, but not all, restriction factors to further curb recurrent cycles of infection. Cell-specific intrinsic immune mechanisms that orchestrate the interferon response are likely to play a key role in the prevention of HIV- 1 infection in long- lived memory CD4+ T-cell subsets.
  • TCM central memory compartment
  • IFITM1 expressing CD4+ T cells are enriched in the central memory compartment (TCM) of ART-suppressed patients.
  • TCM are the main source of latent virus in natural reservoirs.
  • IFITM1 antibody-dependent cytolysis we were able to obtain efficient killing of latently infected CD4+ T cells by IFITM1 antibody-dependent cytolysis.
  • selective targeting of IFITM1 in the TCM reservoir is employed to identify and destroy latently infected cells.
  • HIV/AIDS antivirals include drugs in several categories that affect different aspects of the viral life cycle: viral entry inhibitors, nucleotide/side reverse transcription inhibitors (NRTI), non-nucleoside reverses transcriptase inhibitors (NNRTI), integrase inhibitors and protease inhibitors.
  • NRTI nucleotide/side reverse transcription inhibitors
  • NRTI non-nucleoside reverses transcriptase inhibitors
  • integrase inhibitors integrase inhibitors and protease inhibitors.
  • Combination therapies are variously called anti-retroviral therapy (ART), combination anti- retroviral therapy (cART) or highly active anti-retroviral therapy (HAART).
  • Drugs affecting viral entry include maraviroc (SELZENTRY®), which blocks the CCR5 receptor on helper T-cells, and enfuvirtide (FUZEON®), which is an injectable peptide drug that prevents infection of host cells by interacting with the N-terminal heptad repeat of HIV gp41 to form an inactive hetero six-helix bundle.
  • maraviroc SELZENTRY®
  • FUZEON® enfuvirtide
  • Nucleoside (tide) reverse transcriptase inhibitors act as substrates and competitively inhibit the action of the viral reverse transcriptase and include zidovudine, a.k.a. azidothymidine (RETROVIR®), abacavir (ZIAGEN®), lamivudine (EPIVIR®), emtricitabine (EMTRIVA®), and tenofovir (VIREAD®).
  • Non-nucleoside reverse transcriptase inhibitors bind to the RT and thus inhibit its action.
  • NNRTI include nevirapine (VIRAMUNE®), efavirenz (SUSTIVA®), etravirine (INTELENCE®) and rilpivirine (EDURANT®).
  • Drugs that block HIV integration into the host chromosome act by inhibiting the viral integrase enzyme and include raltegravir (ISENTRESS ® ), elvitegravir (VITEKTA ® ) and dolutegravir (TIVICAY ® ).
  • HIV protease inhibitors include ritonavir (NORVIR ® ), tipranavir (APTIVUS ® ), atazanavir (REYATAZ ® ), darunavir (PREZISTA ® ), indinavir (CRIXIVAN ® ), nelfmavir (VIRACEPT ® ), saquinavir (INVIRASE ® ), lopinavir, amprenavir (AGENERASE ® ) and its prodrug fosamprenavir (LEXIVA ® ).
  • NK cells are large CD3 " CD56 + lymphocytes that are further segregated into CD56 bri ht and CD56 dim subsets. Approximately 90% of peripheral blood and spleen NK cells are CD56 dim and CD16 + with marked cytotoxic activity against target cells. The majority of NK cells found in lymph nodes and tonsils are CD56 bnght CD16 " and perform immune regulation through production of cytokines such as interferon (IFN)-y in response to IL-12, IL-15 and IL-18 stimulation.
  • IFN interferon
  • NK cells are activated and contribute to the initial control of HIV- 1 replication.
  • autologous NK cells from ART- suppressed HIV-1 patients showed limited capacity to produce IFN- ⁇ and to upregulate CD 107a in the presence of anti-IFITMl antibody.
  • the function of NK cells in the presence of an anti-IFITMl antibody is enhanced using stimulation with one or more of type I IFNs (mainly IFN-a and IFN- ⁇ ), IL-2/anti-IL-2 monoclonal antibody complexes, IL-12, IL-15 and IL-18.
  • the function of NK cells in the presence of an anti-IFITMl antibody is enhanced by administration of IL-15 superagonists.
  • IL-15 superagonist is ALT-803, which is an IL-15 mutant (IL-1 5N72D) bound to an IL-15 receptor ⁇ /IgGI Fc fusion protein developed by Altor Bioscience, and has been shown to potently activate NK cells, induce XFN- ⁇ secretion by NK cells and enhance antibody-dependent cell-mediated cytotoxicity (A.DCC) of antibodies in various experimental models.
  • Another II.- 15 super agonist is the fusion protein RLI, composed of the Li-terminal (amino acids 1-77, sushi+) domain of IL-15 receptor a coupled via a linker to IL-15.
  • RLI fusion protein
  • an antibody directed to IFITM1 and including an IgG Fc domain that binds to FcyRl receptors on K cells is administered to an HIV-1 positive patient.
  • the IFITM1 specific antibody supports ADCC mediated killing of cells bearing latent HIV.
  • the antibody is humanized.
  • the antibody is specific to IFITM1 and is not cross reactive with IFITM2 or IFITM3.
  • a monoclonal antibody to IFITM1 is utilized that includes an Fc domain that is selected for its effectiveness in fixing complement and promoting complement-dependent cytotoxicity (CDC) of antibody opsinized cells expressing IFITM1.
  • CDC complement-dependent cytotoxicity
  • antibodies of the IgG3 and IgGl classes are preferred for complement activation through the classic cascade.
  • a monoclonal antibody to IFITM1 is utilized that is optimized for ADCC by directed evolution with selection for increased killing of latently infected CD4+ cells.
  • the antibody is a bi-specific antibody that includes an anti- IFITMl moiety in addition to another moiety having a different target.
  • the different target is CD4.
  • the antibody is a chimeric molecule and is conjugated to a heterologous polypeptide.
  • the IFITM1 antigen binding aspect of the antibody may be a Fab, Fv or antibody half molecule.
  • the heterologous protein is a therapeutic agent.
  • the therapeutic agent is a cytotoxic agent selected from the group consisting of paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, a glucocorticoid, procaine, tetracaine, lidocaine, propranolol, puromycin, epirubicin, or cyclophosphamide.
  • the IFITM1 expressing cells are targeted for cytotoxic killing.
  • an anti-IFITMl specific monoclonal antibody is administered in addition to one or more anti-HIV agents selected from viral entry inhibitors, nucleotide/side reverse transcription inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI), integrase inhibitors and protease inhibitors.
  • an anti-IFITMl specific monoclonal antibody is administered in addition to one or more of type I IFNs, IL-2/anti-IL-2 monoclonal antibody complexes, IL- 12, IL-15 and IL-18.
  • an anti-IFITMl specific monoclonal antibody is administered in addition to an IL-15 superagonist.
  • the IL-15 superagonist is selected from the group cons sting of RLI and ALT-803.
  • an infusion comprising an anti-IFITMl specific monoclonal antibody and an IL-15 superagonist is administered to an HIV-1 patient on ART or HAART therapy.
  • IFITM1 Specific Chimeric Antigen Receptor (CAR) T cells in which immune effector T cells are modified to express an engineered T cell receptor having IFITMl specificity.
  • the CAR includes an IFITM1 specific exodomain formed by antibody VH and VL antigen binding domains separated by a linker to form a fusion protein termed a single-chain variable fragment (scFv) that is specific for binding to IFITMl .
  • scFv single-chain variable fragment
  • a bi-specific CAR T cell that includes an IFITMl specific scFv joined in tandem to a CD4 specific scFv in the CAR exo (extracellular) domain.
  • scFv exodomain(s) are linked by a spacer to a transmembrane domain and an endo (intracellular) signaling domain. See e.g. G Dotti, et al. "Design and Development of Therapies using Chimeric Antigen Receptor-Expressing T cells” Immunol Rev 257(1) (2014) 1-35.
  • the signaling endodomain may include one or more of signaling and co-stimulatory domains derived from CD2, CD3, CD 27, CD28, CD30 (aka TNFRSF8), CD40 (aka TNFRSF5), CD] 34 (aka TNFRSF4 and OX40), CD137 (4- 1BB), CD278 (ICOS, Inducible T-Cell CoStimulator), glucocorticoid-induced TNFR-related protein (GITR; also known as T FRSF18) and combinations thereof.
  • signaling and co-stimulatory domains derived from CD2, CD3, CD 27, CD28, CD30 (aka TNFRSF8), CD40 (aka TNFRSF5), CD] 34 (aka TNFRSF4 and OX40), CD137 (4- 1BB), CD278 (ICOS, Inducible T-Cell CoStimulator), glucocorticoid-induced TNFR-related protein (GITR; also known as T FRSF18) and combinations thereof.
  • the exodomain(s) may be linked to a CD3 ⁇ (CD3-zeta) signaling domain of the T Cell Receptor complex, while in other embodiments the exodomain(s) may be linked to a CD28 ⁇ (zeta) signaling endodomain, in addition to co-stimulatory domains.
  • the CAR T-cell signaling endodomain includes an intracellular signaling domain (CD3 ⁇ ) of the native T cell receptor complex including either intracellular signaling domains of CD28 or 4- IBB as costimulatory domains.
  • the CAR T-Cell endodomain combines CD3 ⁇ to provide cytotoxicity, a Pi3K binding domain derived from a CD28 ⁇ (zeta) signaling endodomain to confer proliferation and cytokine production, and a TRAF2 binding site from a 4-1BB or OX40 intracellular domain to augment survival of the CAR T cell.
  • IL-15 is used to expand the engineered CAR T-cells ex vivo and may be further administered to the recipient patient to support growth and expansion of the engineered CAR T-cells once administered to the patient. Further support for the CAR T-cells may be provided by co-administering a drug that targets one or more immune checkpoint (IC) molecules selected from Programed Cell Death- 1 (PD-1), Programed Cell Death Ligand-1 (PD-L1), T-cell Immunoregulator with Ig and ITIM domains (TIGIT), and Lymphocyte Activation Gene 3 (LAG-3).
  • IC immune checkpoint
  • PD-1 Programed Cell Death- 1
  • PD-L1 Programed Cell Death Ligand-1
  • TAGIT T-cell Immunoregulator with Ig and ITIM domains
  • LAG-3 Lymphocyte Activation Gene 3
  • the IC molecule PD-1 may be inhibited by ivolumab, Pembrolizumab, and Pidilizumab.
  • the IC molecule PD-L1 may be targeted by Atezolizumab and Aveluniab.
  • the IC molecule LAG-3 may be targeted by BMS-986016 and GSK2831781
  • the IC molecule TIGIT may be targeted by Genentech's MTIG7192A.
  • CD4 + T-cells were generated beginning with PBMCs from healthy individuals isolated from buffy-coats as shown figuratively in Fig. 1A.
  • CD4 + T-cell were isolated by negative selection (StemCell).
  • Purified CD4 + T-cells were incubated with CCL19 (29 nM, R&D) for three days before HIV- 1 infection.
  • Cells were infected with HIV-1NL4-3 for 4h at 37°C. The cells were then cultured in the presence of IL-2 (10 IU/ml, Roche) for six days.
  • CD4 + T-cells were isolated by anti-PE magnetic bead depletion (Miltenyi Biotec) using a cocktail of antibodies to CD25-PE (BD Pharmingen, clone M-A251), CD69-PE (BD Pharmingen, clone FN50) and ULA-DR-PE (BD Pharmingen, clone G46-6). Resting latent CD4+ T-cells were then reactivated with anti-CD3/CD28 (StemCell) for three days in the presence of IL-2 (50 IU/ml), according to the manufacturer's specifications.
  • IL-2 50 IU/ml
  • ACH-2 cell line and reactivation with histone deacetylase inhibitors ACH-2 cells were obtained from the NIH AIDS Reagents (catalogue #349) and were tested negative for mycoplasma contamination. Cells were typically propagated in RPMI1640, 2 mM L- Glutamine, 10% FBS in the presence of penicillin/streptomycin.
  • HDACi histone deacetylase inhibitors
  • SAHA suberanilohydroxamic acid
  • Vorinostat marketed by Merck as ZOLINZA®
  • panobinostat marketed under the tradename FARYDAK®
  • romidepsin marketed under the tradename ISTODAX®
  • Cells were typically plated in 96-well plates (U-shape) and treated with different concentrations of FIDACi for 5h, washed twice and re-cultured for 18h prior to RNA extraction or flow cytometry.
  • CD4 + T-cells were washed in FACS buffer (2 mM EDTA, 0.5% BSA in ice-cold PBS) and surface stained with antibodies against CD3 (1 : 100, ECD, clone UCHT1, Beckman Coulter), CD4 (1 :500, Alexa700, clone RPA-T4, Becton Dickinson), CD 8 (1 :400, BV605, clone RPA-T8, Biolegend), and a live/dead marker (1 :200, Invitrogen) emitting in the aqua wavelength, for 30 min at 4°C.
  • PBMCs were washed in FACS buffer and surface stained with monoclonal anti-IFITMl antibody (1 :200, 60074-1-Ig, monoclonal mouse anti-human, Proteintech) for 30 min at 4°C. After staining, cells were washed three times and stained with anti-mouse IgG2a-PE (1 :200, Biolegend).
  • PBMCs were washed and stained for CD3 (1 : 100, ECD, clone UCHT1, Beckman Coulter), CD4 (1 :500, Alexa700, clone RPA-T4, Becton Dickinson), CD8 (1 :400, BV605, clone RPA-T8, Biolegend), CD45RA (1 : 100, BV650, clone HI100, BD Pharmingen), CCR7 (1 : 100, PE-Cy7, cloneG043H7, Biolegend), CD27 (1 : 100, APC-Cy7, clone 0323, Biolegend) and CD95 (1 :50, PacificBlue, clone DX2, Biolegend) and a live/dead marker (1 :200, Invitrogen) for 30 min at 4°C.
  • CD3 1 : 100, ECD, clone UCHT1, Beckman Coulter
  • CD4 (1 :500, Alexa700,
  • CD4+ T cells were incubated for 30min at 4°C with anti-IFITMl antibody (1 :200, 60074-1 -Ig, Proteintech) and after staining, cells were washed three times and stained with anti-mouse IgG2a-PE (1 :200, Biolegend) for an additional 30min at 4°C in the dark.
  • Anti-PE microbeads (Miltenyi Biotec) were used to deplete IFITM1+ cells from bulk CD4+ T cells.
  • IFITM1-CD4+ T cells were collected and genomic DNA was isolated using DNeasy Blood & Tissue kit (Qiagen), according to the manufacture's recommendations.
  • ADCC and functional assays ACH-2 cells (NIH AIDS Reagents) or CD4 + T-cells, isolated from PBMC of ART-suppressed patients by negative selection (StemCell), were incubated with monoclonal anti-IFITMl antibody (1 :200, 60074-1-Ig, Proteintech) or IgG2a isotype control (Biolegend) prior to the addition of PBMC effector cells for 5h at the indicated effector to target ratio (20: 1 and 10: 1, in a 96-well U-shaped plate in a final volume of 200 ⁇ ). Autologous PBMCs or heterologous PBMCs from healthy donors were used as effector cells.
  • Monensin (Golgi Stop, BD Biosciences) was added at the beginning of the assay.
  • Target cell apoptosis was detected using the fluorescent inhibitor of caspases (FLIC A) flow cytometry -based assay as described by Leeanshya, E. et al. Arming of MAIT Cell Cytolytic Antimicrobial Activity Is Induced by IL-7 and Defective in HIV-1 Infection. PLoS Pathog. 11(8) (2015) el005072. Briefly, the FLICA reagent (Vybrant FAM Poly-Caspases Assay Kit, Thermofisher) was added in the last hour to the cell culture media.
  • FLICA reagent Vybrant FAM Poly-Caspases Assay Kit, Thermofisher
  • CD4+ T cells were isolated from PBMC of a ART- suppressed patient by negative selection (StemCell) and incubated with monoclonal anti- IFITM1 antibody (1 :200, 60074-1-Ig, Proteintech) or IgG2a isotype control (Biolegend) prior to the addition of autologous PBMC as effector cells for three days at an effector to target ratio of 10.
  • CD4+ T cells were re-purified by negative selection and stimulated with PHA (lOug/ml) and IL-2 (50U/ml) for an addition three days. Cells were harvested and cell-associated RNA was extracted using TRIzol reagent.
  • RNA from CD4 + T-cells was extracted using TRIzol reagent, followed by RNACleanup with in solution DNAasel treatment option using Qiagen RNase-Free DNase Set.
  • DNase-treated clean RNA was transcribed into cDNA using random primers and the Superscript VILO cDNA Synthesis Kit (Invitrogen), according to manufacturer's instructions.
  • Quantitative real-time PCR on blinded samples utilized custom-made TaqMan Low Density Array (TLDA) from Applied Biosystems and gene cards used for quantification as first developed in our laboratory. See Abdel-Mohsen M, et al. Expression profile of host restriction factors in HIV-1 elite controllers.
  • HIV-1 Gag and mRNA transcripts were detected using the primers and probe described in Shan L, et al. A novel PCR assay for quantification of HIV-1 RNA. J Virol. 87(11) (2013) 6521-6525. CCR5 (Life Technologies, Hs99999149_sl) was used as endogenous control. Real-time PCR was performed in duplicate using TaqMan Universal PCR Master Mix (Applied Biosystems) on an ABI ViiA7 Real-Time PCR machine. Fold induction was determined using the delta delta Ct method. [0079] Statistical analysis: Statistical comparisons were performed using non-parametric two-tailed t-tests in GraphPad Prism. Data are plotted as means ⁇ SEM or SD and P ⁇ 0.05 were considered significant.
  • HIV-1 latent cells were generated using an ex vivo model as depicted generally in Fig. 1A using the HIV-1+ molecular clone HIV-1.
  • CD4+ T-cells were isolated from healthy donors, conditioned for three days in the presence of CCL19 and then infected with HIV-1.
  • the initial absence of HIV-1 Gag protein was confirmed by flow cytometry (KC57 antibody).
  • the KC57 monoclonal antibody available from Becton Coulter identifies the 55, 39, 33 and 24 kD proteins of the core antigens of Human Immunodeficiency Virus Type 1 (HIV-1).
  • the 55 kD protein is the precursor protein for the core antigen.
  • the 39 and 33 kD proteins are intermediate products and the 24 kD protein is the mature core protein that makes up the viral capsids.
  • the 24 kD core protein is also known as the HIV gag (group- specific antigen) p24 protein.
  • Fig. IB shows HIV-1 gag staining at day 6 post-infection in resting (CD25-, CD69- and HLA-DR-) latent CD4 + T-cells. Latent HIV-1 was then reactivated in resting cells with anti-CD3/CD28 for three days with a resulting dramatic increase in HIV-1 gag staining by flow cytometry as shown in Fig. 1C.
  • Figs. 2A-C As sown in Figs. 2A-C, reactivation led to a 10-fold induction in HIV-1 gag (Fig. 2A) and HIV-1 mRNA transcripts (Fig. 2B), including unspliced (US), single spliced (SS) and multiple spliced (MS) mRNA, as measured by real-time qPCR.
  • the expression of selected antiviral genes and host restriction factors was comprehensively analyzed and found that the expression of IFITM1 and SAMHDl were 4-fold and 2-fold increased, respectively, in resting latent cells when compared to reactivated cells (Fig. 2C).
  • Figs. 6A - G depict the results of reactivation of HIV-1 in ACH-2 cells.
  • HDACi histone deacetylase inhibitors
  • SAHA is the acronym for suberanilohydroxamic acid, which is referred to by the generic term Vorinostat and is marketed by Merck under the tradename ZOLINDA®.
  • Vorinostat is FDA approved for treatment of cutaneous T cell lymphoma (CTCL) but was recently shown to have both in vitro and in vivo effects against latently HIV infected T cells by inducing reactivation from latency in chronically infected cell lines and primary cells. See Contreras, X, et al. "Suberoylanilide Hydroxamic Acid Reactivates HIV from Latently Infected Cells" JBC 284 (11) (2009) 6782-6789.
  • Fig. 6G shows the effect of different HDACi on ACH-2 cell viability. The frequency of IFITM1+ CD4 + T-cells was further confirmed to be reduced upon HIV-1 reactivation (Figs. 3A - D).
  • IFITM1 is a transmembrane protein
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • ACH-2 cells are a T-cell clonal line that is latently infected with HIV-1 and available through the NIH AIDS Reagent Program (Division of AIDS, NIAID, NIH: ACH-2 from Dr. Thomas Folks).
  • ACH-2 cells are CD4-, CD5+, transferrin receptor+, Leu-1+, HIV-1+.
  • ACH-2 cells constantly produce low levels of supernatant reverse transcriptase (RT) and p24.
  • ACH-2 cells can be induced with phorbol myristate acetate or T F- ⁇ to secrete high levels of infectious HIV-1.
  • Latently infected ACH-2 cells were labeled with an anti-IFITMl antibody or control antibody and cultured with PBMCs from healthy donors at different effector to target (E:T) ratios.
  • Dead cells were monitored by flow cytometry using a live/dead cell marker while the early stages of apoptosis were detected using the FAM Poly-Caspases assay kit (FLICA).
  • FLICA FAM Poly-Caspases assay kit
  • a FLICA kit is available from Immunochemistry Technologies LLC and the assay employs the fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase-1 enzyme in living cells or tissue samples and thereby quantitate apoptosis.
  • FIG. 4A Representative flow plots of LIVE/DEAD Aqua vs. FLICA+ gated on the target cells (ACH-2) are depicted in Fig. 4A.
  • ACH-2 target cells
  • NK cell defined as CD3- and CD56+ lymphocytes
  • Representative flow plots are depicted in Fig. 4B.
  • Significant killing of ACH-2 cells labeled with anti-IFITMl antibody was detected when compared to control (25 ⁇ 1.3%) (Fig.
  • Fig. 7A- 7H shows the side scatter area (SSC-A) plotted against the Forward Scatter area (FSC-A) by flow cytometry with the cells to be analyzed within the drawn area.
  • Fig. 7B shows the Forward Scatter area (FSC-A) plotted against the Forward Scatter height (FSC-H) to identify single cells and exclude doublets with cells outside of the box excluded from analysis.
  • PBMC were stained with antibodies against CD3 (Fig. 7C), CD4 (Fig. 7D and F), CD 8 (Fig. 7D), CD45RA (Fig. 7F), CCR7 (Figs. 7E and G), CD27 (Figs. 7E and G), CD95 (Fig. 7H) and IFITM1.
  • Dead cells were excluded using a live/dead marker.
  • naive cells are CD3 + CD4 + CD45RA + CD27 + CCR7 + CD95 " .
  • Stem central memory (TSCM) cells are also CD3 + CD4 + CD45RA + CD27 + CCR7 + CD95 " .
  • Central memory (TCM) cells are CD3 + CD4 + CD45RA " CD27 + CCR7 + .
  • Transitional memory (TTM) cells are CD3 + CD4 + CD45RA " CD27 + CCR7 " .
  • Effector memory (TEM) cells are CD3 + CD4 + CD45RA + CD27 " CCR7 " .
  • TABLE 1 presents the population identifiers in tabular form.
  • IFITM1 + CD4 + T-cells were killed by ADCC, CD4 + T-cells from ART-suppressed patients were sorted, labeled with anti-IFITMl antibody, and cultured with effector cells from the same patient for 5h. IFITM1 + T-cells were found almost exclusively in the fraction positive for amine-reactive live/dead marker and FLICA (Fig. 5B), suggesting efficient and specific killing.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Hematology (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • AIDS & HIV (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne des procédés et des compositions pour identifier des cellules infectées de façon latente par et hébergeant un réservoir occulte de virus infectieux. Dans certains modes de réalisation, les taux d'expression d'IFITM1 par les lymphocytes T sont surveillés pour évaluer l'efficacité d'un traitement pour le VIH. Dans d'autres modes de réalisation, le traitement du VIH latent est conduit par ciblage de cellules exprimant IFITM1 pour une cytolyse médiée par anticorps. Dans d'autres modes de réalisation, des lymphocytes T CAR ayant une spécificité monospécifique pour IFITM1 ou bispécifique pour IFITM1 et CD4 sont générés et utilisés pour traiter des individus infectés par le VIH.
PCT/US2017/026669 2016-04-07 2017-04-07 Procédés et compositions ciblant une latence rétrovirale Ceased WO2017177175A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201662319668P 2016-04-07 2016-04-07
US62/319,668 2016-04-07

Publications (1)

Publication Number Publication Date
WO2017177175A1 true WO2017177175A1 (fr) 2017-10-12

Family

ID=60001545

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2017/026669 Ceased WO2017177175A1 (fr) 2016-04-07 2017-04-07 Procédés et compositions ciblant une latence rétrovirale

Country Status (1)

Country Link
WO (1) WO2017177175A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020077180A1 (fr) * 2018-10-11 2020-04-16 Nantcell, Inc. Traitement de sujets immunodéprimés
WO2020228824A1 (fr) * 2019-05-16 2020-11-19 Nanjing Legend Biotech Co., Ltd. Récepteurs de cellules immunitaires comprenant des fractions de liaison à cd4

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070041981A1 (en) * 2005-06-01 2007-02-22 Northwestern University Compositions and methods for altering immune function
US20110275542A1 (en) * 2010-04-21 2011-11-10 MeMed Diagnostics, Ltd. Signatures and determinants for distinguishing between a bacterial and viral infection and methods of use thereof
US20140242075A1 (en) * 2011-05-30 2014-08-28 Genmab B.V. Antibody variants and uses thereof
US20150024482A1 (en) * 2012-02-22 2015-01-22 The Trustees Of The University Of Pennsylvania Compositions and Methods for Generating a Persisting Population of T Cells Useful for the Treatment of Cancer
WO2015121454A1 (fr) * 2014-02-14 2015-08-20 Cellectis Cellules destinées à l'immunothérapie modifiées pour cibler un antigène présent à la fois sur les cellules immunitaires et les cellules pathologiques
US20150374790A1 (en) * 2014-06-30 2015-12-31 Altor Bioscience Corporation Il-15-based molecules and methods of use thereof
US20160030659A1 (en) * 2013-03-15 2016-02-04 Novelogics Biotechnology, Inc. Methods and devices for removal of immunosuppressive ligands

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070041981A1 (en) * 2005-06-01 2007-02-22 Northwestern University Compositions and methods for altering immune function
US20110275542A1 (en) * 2010-04-21 2011-11-10 MeMed Diagnostics, Ltd. Signatures and determinants for distinguishing between a bacterial and viral infection and methods of use thereof
US20140242075A1 (en) * 2011-05-30 2014-08-28 Genmab B.V. Antibody variants and uses thereof
US20150024482A1 (en) * 2012-02-22 2015-01-22 The Trustees Of The University Of Pennsylvania Compositions and Methods for Generating a Persisting Population of T Cells Useful for the Treatment of Cancer
US20160030659A1 (en) * 2013-03-15 2016-02-04 Novelogics Biotechnology, Inc. Methods and devices for removal of immunosuppressive ligands
WO2015121454A1 (fr) * 2014-02-14 2015-08-20 Cellectis Cellules destinées à l'immunothérapie modifiées pour cibler un antigène présent à la fois sur les cellules immunitaires et les cellules pathologiques
US20150374790A1 (en) * 2014-06-30 2015-12-31 Altor Bioscience Corporation Il-15-based molecules and methods of use thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JIA ET AL.: "The C-Terminal Sequence of IFITM1 Regulates Its Anti-HIV-1 Activity", PLOS ONE, vol. 10, no. 3, 4 March 2015 (2015-03-04), pages 1 - 17, XP055430281 *
JOHNSTON ET AL.: "The immunoreceptor TIGIT regulates antitumor and antiviral CD 8(+) T cell effector function", CANCER CELL, vol. 26, no. 6, 8 December 2014 (2014-12-08), pages 923 - 937, XP029111576 *
YU ET AL.: "IFITM Proteins Restrict HIV-1 Infection by Antagonizing the Envelope Glycoprotein", CELL REPORTS, vol. 13, no. 1, 17 September 2015 (2015-09-17), pages 145 - 156, XP055430286 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020077180A1 (fr) * 2018-10-11 2020-04-16 Nantcell, Inc. Traitement de sujets immunodéprimés
US11701408B2 (en) 2018-10-11 2023-07-18 Nantcell, Inc. Treatment of immunosuppressed subjects
US12156901B2 (en) 2018-10-11 2024-12-03 Nantcell, Inc. Treatment of immunosuppressed subjects
US12303552B2 (en) 2018-10-11 2025-05-20 Nantcell, Inc. Treatment of immunosuppressed subjects
US12427184B2 (en) 2018-10-11 2025-09-30 Nantcell, Inc. Treatment of immunosuppressed subjects
WO2020228824A1 (fr) * 2019-05-16 2020-11-19 Nanjing Legend Biotech Co., Ltd. Récepteurs de cellules immunitaires comprenant des fractions de liaison à cd4

Similar Documents

Publication Publication Date Title
Van der Sluis et al. Diverse effects of interferon alpha on the establishment and reversal of HIV latency
US20250041401A1 (en) Methods of identifying hiv patients sensitive to therapy with gp120 v3 glycan-directed antibodies
Ellegård et al. Impaired NK cell activation and chemotaxis toward dendritic cells exposed to complement-opsonized HIV-1
WO2017177175A1 (fr) Procédés et compositions ciblant une latence rétrovirale
Bernstein et al. Immune reconstitution following autologous transfers of CD3/CD28 stimulated CD4+ T cells to HIV-infected persons
JP2024107366A (ja) ｇｐ１２０ ＣＤ４結合部位指向性抗体による治療に感受性のＨＩＶ患者を特定する方法
US20160108122A1 (en) Therapeutic or prophylactic agent for immunodeficiency virus infection
Pan et al. Allogeneic gene-edited HIV-specific CAR-T cells secreting PD-1 blocking scFv enhance anti-HIV activity in vitro
Diaz Impact of early antiretroviral therapy initiation and immunotherapy interventions on the dynamics of regulatory T cells during SIV-HIV infections
Debaveye Generation and efficacy analysis of a bispecific nanobody targeting CD8+ T cells and HIV-infected cells
Laeremans et al. From natural defenders to therapeutic warriors: NK cells in HIV immunotherapy
JP2025175053A (ja) Ｇｐ１２０ ｖ３グリカン指向性抗体による療法に感受性であるｈｉｖ患者を同定する方法
EP4138910A1 (fr) Nouvelle combinaison pour le traitement du sida
HK40070195B (en) Methods of identifying hiv patients sensitive to therapy with gp120 v3 glycan-directed antibodies
HK40070195A (en) Methods of identifying hiv patients sensitive to therapy with gp120 v3 glycan-directed antibodies
HK40061191A (en) Methods of identifying hiv patients sensitive to therapy with gp120 v3 glycan-directed antibodies

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17779934

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17779934

Country of ref document: EP

Kind code of ref document: A1