[go: up one dir, main page]

CN111574615A - TCR enrichment clone type and acquisition method and application thereof - Google Patents

TCR enrichment clone type and acquisition method and application thereof Download PDF

Info

Publication number
CN111574615A
CN111574615A CN202010445718.7A CN202010445718A CN111574615A CN 111574615 A CN111574615 A CN 111574615A CN 202010445718 A CN202010445718 A CN 202010445718A CN 111574615 A CN111574615 A CN 111574615A
Authority
CN
China
Prior art keywords
tcr
clonotype
enriched
enrichment
obtaining
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.)
Pending
Application number
CN202010445718.7A
Other languages
Chinese (zh)
Inventor
伊宁
薛志刚
薛金锋
李婵艺
齐凌彬
吕波
李伟林
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.)
Hunan Yuanpin Cell Biotechnology Co ltd
Original Assignee
Hunan Yuanpin Cell Biotechnology Co ltd
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 Hunan Yuanpin Cell Biotechnology Co ltd filed Critical Hunan Yuanpin Cell Biotechnology Co ltd
Priority to CN202010445718.7A priority Critical patent/CN111574615A/en
Priority to PCT/CN2020/105806 priority patent/WO2021237932A1/en
Publication of CN111574615A publication Critical patent/CN111574615A/en
Priority to CN202110564597.2A priority patent/CN113214383A/en
Priority to US17/475,686 priority patent/US12252526B2/en
Pending legal-status Critical Current

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503Immunoglobulin superfamily
    • C07K14/7051T-cell receptor (TcR)-CD3 complex
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06Animal cells or tissues; Human cells or tissues
    • C12N5/0602Vertebrate cells
    • C12N5/0634Cells from the blood or the immune system
    • C12N5/0636T lymphocytes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2509/00Methods for the dissociation of cells, e.g. specific use of enzymes

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Toxicology (AREA)
  • General Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a TCR enrichment clonotype and an acquisition method and application thereof, wherein the amino acid sequence of the TCR enrichment clonotype is CAVNSYNTDKLIF _ CATSREEDNTYEQYF, which can provide antigen epitope for vaccine development and specific recognition, so that the TCR enrichment clonotype can be used for guiding vaccine development and providing direction and theoretical basis for vaccine development and research.

Description

TCR enrichment clone type and acquisition method and application thereof
Technical Field
The invention relates to the technical field of biomedicine, in particular to a TCR enrichment clonotype and an acquisition method and application thereof.
Background
The outbreak of new coronavirus (2019-novel coronavirus, 2019-nCoV) infection poses a serious threat to global public health. The disease caused by 2019-nCoV is formally named COVID-19(coronavirus disease 2019) by World Health Organization (WHO). COVID-19 clinically manifests as pneumonia, fever, cough, muscle pain, fatigue, diarrhea, and death in severe cases. The WHO report shows that 2019-nCoV has caused a world pandemic. Vaccination is an effective means for preventing viral infection epidemics, but there is no new coronavirus vaccine (hereinafter referred to as new coronavirus vaccine) at present. At present, the treatment means mainly supports nursing aiming at the COVID-19 virus which is rapidly spread by public, and antiviral drugs, antibodies, vaccines and the like aiming at the COVID-19 are still in the clinical experiment stage and are not successfully developed. The same method as SARS in the current year is mostly adopted for symptomatic treatment, and the inflammatory reaction caused by the immune system is suppressed by hormone, but the human body is also very fragile and difficult to be protected from injury. There is an urgent need for effective vaccine development to contain the outbreaks of viruses worldwide.
T cell (antigen) receptors (TCRs), which are characteristic markers of all T cell surfaces, bind to CD3 by non-covalent bonds, forming a TCR-CD 3 complex. The role of the TCR is to recognize an antigen. The TCR is a heterodimer formed by two different peptide chains and consists of alpha and beta peptide chains, and each peptide chain can be divided into a variable region (V region), a constant region (C region), a transmembrane region, a cytoplasmic region and the like; characterized by a short cytoplasmic domain. The TCR molecule belongs to the immunoglobulin superfamily, and the antigen specificity exists in the V region; the V regions (V α, V β) in turn have three hypervariable regions CDR1, CDR2, CDR3, of which the largest variation in CDR3 directly determines the antigen binding specificity of the TCR. When the TCR recognizes the MHC-antigen peptide complex, the CDRs 1, 2 recognize and bind to the side walls of the antigen binding groove of the MHC molecule, while the CDR3 binds directly to the antigen peptide. The TCR enrichment clonotypes provided by the invention have positive effects on specific recognition of antigen epitopes through the common CDR3 variable region, and can be used for guiding vaccine development and providing direction and theoretical basis for vaccine development and research.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a TCR enrichment clonotype and an acquisition method and application thereof.
In order to achieve the above object, the present invention discloses a TCR-enriched clonotype having an amino acid sequence of CAVNSYNTDKLIF _ CATSREEDNTYEQYF.
The invention also discloses a method for acquiring the TCR enrichment clonotype, which comprises the following steps:
s1, collecting a peripheral blood mononuclear cell sample of the cured virus infected patient;
s2, carrying out TCR/BCR V (D) J immune repertoire sequencing analysis on the collected sample, and finding out a common CDR3 variable region;
s3, detecting the cell clonotype distribution with high enrichment degree of the cured virus infected patient, thereby obtaining the TCR enrichment clonotype.
Wherein the TCR-enriched clonotypes are enriched in cell clusters of effector CD8+ T cells.
The invention also discloses application of the TCR enrichment clonotype in guiding vaccine development.
Wherein the vaccine is 2019-nCoV vaccine.
The beneficial technical effects are as follows: the amino acid sequence of the TCR enrichment clonotype is CAVNSYNTDKLIF-CATSREEDNTYEQYF, and the TCR enrichment clonotype has positive significance on the antigen epitope specifically identified through a common CDR3 variable region, so that the TCR enrichment clonotype can be used for guiding vaccine development and provides a direction and theoretical basis for vaccine development and research.
Drawings
FIG. 1 is a flow chart of a method for obtaining TCR-enriched clonotypes of the invention,
FIG. 2 is a schematic representation of the location of TCR-enriched clonotypes of the invention and their degree of enrichment in TCR cells.
Detailed Description
In order to make the technical solutions of the present invention better understood, the present invention is further described in detail below with reference to the accompanying drawings.
The invention provides a TCR enrichment clonotype, and the amino acid sequence of the TCR enrichment clonotype is CAVNSYNTDKLIF-CATSREEDNTYEQYF.
As shown in fig. 1 and fig. 2, in this embodiment, the method for obtaining a TCR-enriched clonotype specifically includes the following steps:
s1, collecting a peripheral blood mononuclear cell sample of the cured virus infected patient;
s2, carrying out TCR/BCR V (D) J immune repertoire sequencing analysis on the collected sample, and finding out a common CDR3 variable region;
s3, detecting the distribution of cell clonotypes with high enrichment degree of the cured virus infected patient, thereby obtaining the TCR enrichment clonotypes.
In the embodiment, single nuclear cells of peripheral blood of four 2019-nCoV virus infected patients are selected as sampling samples, then TCR/BCR V (D) J immune repertoire sequencing analysis is carried out on the sampling samples, although the sequencing analysis result has no common cell clonotypes, a common CDR3 variable region exists, and the TCR enrichment clonotypes enriched in effector CD8+ T cell clusters are obtained by detecting the cell clonotype distribution with high enrichment degree of the virus infected patients, so that the TCR enrichment clonotypes have positive effect on specific recognition antigen epitopes through the common CDR3 variable region.
In this embodiment, the TCR-enriched clonotypes described above are applied to guide vaccine development, particularly to guide 2019-nCoV vaccine development, and since the TCR-enriched clonotypes have a positive effect on specific recognition of antigenic epitopes through the common CDR3 variable region, the TCR-enriched clonotypes can be used for guiding vaccine development and provide a direction and theoretical basis for vaccine development and research.
The TCR-enriched clonotypes provided by the invention, the method for obtaining the same and the application thereof are described in detail above. The principles and embodiments of the present invention are explained herein using specific examples, which are presented only to assist in understanding the core concepts of the present invention. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.

Claims (5)

1. A TCR-enriched clonotype having the amino acid sequence CAVNSYNTDKLIF _ CATSREEDNTYEQYF.
2. A method of obtaining a TCR-enriched clonotype as claimed in claim 1, comprising the steps of:
s1, collecting a peripheral blood mononuclear cell sample of the cured virus infected patient;
s2, carrying out TCR/BCR V (D) J immune repertoire sequencing analysis on the collected sample, and finding out a common CDR3 variable region;
s3, detecting the distribution of cell clonotypes with high enrichment degree of the cured virus infected patient, and further obtaining the TCR enrichment clonotypes.
3. A method of obtaining a TCR-enriched clonotype as claimed in claim 2 wherein the TCR-enriched clonotype is enriched in a cell cluster of effector CD8+ T cells.
4. Use of a TCR-enriched clonotype according to claim 1 or a TCR-enriched clonotype obtained by the method of obtaining a TCR-enriched clonotype according to claim 2 or 3 for directing vaccine development.
5. Use of a TCR-enriched clonotype as claimed in claim 4 for directing vaccine development, wherein the vaccine is a 2019-nCoV vaccine.
CN202010445718.7A 2020-05-23 2020-05-23 TCR enrichment clone type and acquisition method and application thereof Pending CN111574615A (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
CN202010445718.7A CN111574615A (en) 2020-05-23 2020-05-23 TCR enrichment clone type and acquisition method and application thereof
PCT/CN2020/105806 WO2021237932A1 (en) 2020-05-23 2020-07-30 Tcr enriched clonotypes and acquisition method and application therefor
CN202110564597.2A CN113214383A (en) 2020-05-23 2021-05-24 TCR enrichment clone type and acquisition method and application thereof
US17/475,686 US12252526B2 (en) 2020-05-23 2021-09-15 TCR-enriched clonotype, acquisition method and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010445718.7A CN111574615A (en) 2020-05-23 2020-05-23 TCR enrichment clone type and acquisition method and application thereof

Publications (1)

Publication Number Publication Date
CN111574615A true CN111574615A (en) 2020-08-25

Family

ID=72121291

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202010445718.7A Pending CN111574615A (en) 2020-05-23 2020-05-23 TCR enrichment clone type and acquisition method and application thereof
CN202110564597.2A Pending CN113214383A (en) 2020-05-23 2021-05-24 TCR enrichment clone type and acquisition method and application thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN202110564597.2A Pending CN113214383A (en) 2020-05-23 2021-05-24 TCR enrichment clone type and acquisition method and application thereof

Country Status (1)

Country Link
CN (2) CN111574615A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023138658A1 (en) * 2022-01-24 2023-07-27 卡瑞济(北京)生命科技有限公司 2019-ncov-specific t cell receptor and use thereof

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1609196A (en) * 2003-10-17 2005-04-27 上海普泛生物科技有限公司 High-yield T lymphocyte cloning technology for finding tumor specific antigen and tumor specific antigen determinant
CN101200767B (en) * 2007-12-05 2012-08-22 浙江大学 Method for detecting peripheral blood specific T lymphocyte
CN102212888A (en) * 2011-03-17 2011-10-12 靳海峰 High throughput sequencing-based method for constructing immune group library
US11001830B2 (en) * 2015-03-16 2021-05-11 Max-Delbrück-Centrum Für Molekulare Medizin In Der Helmholtz-Gemeinschaft Method of detecting new immunogenic T cell epitopes and isolating new antigen-specific T cell receptors by means of an MHC cell library
CN106632659A (en) * 2016-08-05 2017-05-10 武汉赛云博生物科技有限公司 EB (Epstein-Barr) virus specific TCR (T cell receptor) and recombinant lentiviral vector and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023138658A1 (en) * 2022-01-24 2023-07-27 卡瑞济(北京)生命科技有限公司 2019-ncov-specific t cell receptor and use thereof

Also Published As

Publication number Publication date
CN113214383A (en) 2021-08-06

Similar Documents

Publication Publication Date Title
Miller Effect of thymectomy in adult mice on immunological responsiveness
Mousavi et al. Recent biotechnological approaches for treatment of novel COVID-19: From bench to clinical trial
CN111574614A (en) TCR enrichment clone type and acquisition method and application thereof
RU2012127380A (en) HUMANIZED ANTIBODIES AGAINST IL-10 FOR TREATMENT OF SYSTEM RED Lupus erythematosus (SLT)
Koponen et al. T cell reactivity to penicillin: phenotypic analysis of in vitro activated cell subsets
Sheng et al. Immune response and safety of heterologous ChAdOx1-nCoV-19/mRNA-1273 vaccination compared with homologous ChAdOx1-nCoV-19 or homologous mRNA-1273 vaccination
Masuko et al. Dynamic changes of accumulated T cell clonotypes during antigenic stimulation in vivo and in vitro
MX2014006248A (en) Monoclonal antibodies specific for the m2-1 antigen of respiratory syncytial virus (rsv).
WO2021237932A1 (en) Tcr enriched clonotypes and acquisition method and application therefor
CN111574615A (en) TCR enrichment clone type and acquisition method and application thereof
Kho et al. Humoral and cellular response after varicella vaccination in VZV IgG seronegative kidney transplant candidates
CN111518195A (en) TCR enrichment clone type and acquisition method and application thereof
CN111574613A (en) TCR enrichment clone type and acquisition method and application thereof
Diamantidis et al. Hemophagocytic lymphohistiocytosis as a manifestation of underlying visceral leishmaniasis
CN111518196A (en) TCR enrichment clone type and acquisition method and application thereof
Davis et al. Immune status of patients with multiple sclerosis: analysis of primary and established immune responses in 24 patients
Kirkpatrick et al. Immunobiology of transfer factor
Al Jadiry et al. High incidence of hepatitis B infection after treatment for paediatric cancer at a teaching hospital in Baghdad
Armistead et al. Mucosal memory T cells in breastmilk are modulated by SARS-CoV-2 mRNA vaccination
CN1480215A (en) Compound antibody vaccine of SARS virus antigen as well as its model of experimental animal and method
Koc et al. Case report of delayed seroprotection rather than non-response after primary three-dose hepatitis B vaccination
CN108314708B (en) Bursal active nonapeptide capable of promoting vaccine immune response and application thereof
Khatoon et al. Prevalence of Transfusion-Transmitted Infections and Its Association With Blood Groups Among Blood Donors at the Blood Bank of a Tertiary Care Center
Doherty The keys to cell-mediated immunity
Gualberto et al. Intermittent rash, lymph node swelling, arthralgia and vaccinal viral detection after rubella immunization

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20200825