CN110016073A - A kind of XAGE-1b nonapeptide and its application - Google Patents

A kind of XAGE-1b nonapeptide and its application Download PDF

Info

Publication number
CN110016073A
CN110016073A CN201910233750.6A CN201910233750A CN110016073A CN 110016073 A CN110016073 A CN 110016073A CN 201910233750 A CN201910233750 A CN 201910233750A CN 110016073 A CN110016073 A CN 110016073A
Authority
CN
China
Prior art keywords
xage
cell
peptide
ctl
small peptide
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
CN201910233750.6A
Other languages
Chinese (zh)
Inventor
张蓓
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.)
Guangzhou Biological Technology Co Ltd
Original Assignee
Guangzhou Biological Technology 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 Guangzhou Biological Technology Co Ltd filed Critical Guangzhou Biological Technology Co Ltd
Publication of CN110016073A publication Critical patent/CN110016073A/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4748—Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
    • 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]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57423—Specifically defined cancers of lung
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5154—Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/11—Coculture with; Conditioned medium produced by blood or immune system cells
    • C12N2502/1121—Dendritic cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • General Physics & Mathematics (AREA)
  • Genetics & Genomics (AREA)
  • Pathology (AREA)
  • Oncology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Hospice & Palliative Care (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Veterinary Medicine (AREA)
  • Toxicology (AREA)
  • General Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Mycology (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)

Abstract

The invention discloses a kind of XAGE-1b nonapeptide and its applications.The sequence of XAGE-1b small peptide is GSRQKKIRI.The CTL that XAGE-1b antigen inducing peptide of the invention is established will not generate immune response to testicular cell, killing tumor cell.Induce obtained ctl clone that there is good tumor cell specific lethal effect.Simultaneously during establishing CTL, the XAGE-1b Antigenic Peptide that filters out of inventor's discovery has and HLA affinity appropriate on DC cell and can effectively stimulate, inducing producing specificity CTLs, illustrates that it has the potentiality of good polypeptide vaccine and DC vaccine.By cloning XAGE-1b Antigenic Peptide specificity TCR gene, construct viral vectors, transduction peripheral circulation CD8+T cell, obtain T cell (TCR-T) clone of tcr gene modification, equally there is tumor cell specific lethal effect similar with parental generation CTL, prompt it with good clinical conversion and actual application prospect.

Description

A kind of XAGE-1b nonapeptide and its application
Technical field
Tumor associated antigen peptide of the present invention and its application, in particular to tumor associated antigen XAGE-1b small peptide and its application.
Background technique
It is well known that T cell is adopted, to treat tumour be that the technology that most attracts people's attention, especially receptor engineering T are thin at present The application of born of the same parents (TCR-T) just becomes soul-stirring cancer treatment method, equally has to virus infection and autoimmune disease Have wide practical use.
Human immunity monitoring system is resisting the important function in Tumor Growth, and immune surveillance system is each by coordinating Para-immunity cell is achieved the purpose for inhibiting tumour growth, wherein cytotoxic T cell (cytotoxic T Lymphocyte, CTLs) play the part of extremely important role, it is the lethal effect cell of most critical.Specific for tumour antigen CD8+T Cell can be with specific recognition and direct killing tumour cell is without damaging normal cell.Therefore T cell treatment becomes Optimal ideas of cancer therapy.U.S. NIH once foretold that cell therapy, which may become, " can thoroughly cure the unique of tumour Means ".Therefore, obtain can specific recognition, killing tumor cell CTL be treat tumour core.
Tumor antigen peptide is established using the induction of tumor associated antigen (Tumor-associated antigen, TAA) small peptide Specific CTL can be obtained the ctl clone with specific killing tumour cell, and have clinical practice for exploitation in next step The T cell of application value, which is adopted, treats tumour and has established important foundation, while herein on basis, can also obtain as follows at Fruit:
1) these pass through the tumor associated antigen peptide amino acid sequence that evaluation and screening goes out, and it is effective anti-will to can be used for building Tumour peptide vaccine, DC vaccine；
2) inventor can be made to obtain the explicitly TCR with specific recognition tumour antigen by molecule clone technology to compile Code gene, the foundation for tcr gene modification T cell；
3) by the fluorescence probe of synthesis and small peptide specific binding, molecular probe with independent intellectual property rights can be developed Kit, early diagnosis and treatment for non-small cell lung cancer.
Currently, T cell receptor (T cell receptor, TCR) gene modification T cell (TCR-T), such as chimeric antigen Receptor (chimeric antigen receptor, CAR) modification T cell (CAR-T) technology equally receives much attention.CAR-T application The curative effect to attract people's attention has been obtained in neoplastic hematologic disorder treatment.CAR merges CD3 signal element by scFv, and functional activity relies on In the sensibility of signal element, signal element is made of costimulatory molecules and (or) cell factor.However, there is studies have shown that table It is not so good as the T cell of express alpha β TCR heterodimer to the sensibility of polypeptide up to the T cell of CAR.TCR-T is repaired by tcr gene T cell technology is adornd, by the acquired TCR encoding gene transduction circulation CD8 with specific recognition tumor-antigen peptide+T is thin Born of the same parents, can be obtained in a very short period of time by this method largely stablize high expression specificity TCR, have and parental generation CTL The gene modification T cell of same tumour-specific identification and lethal effect, to meet needed for practical clinical.Inventor The induction for beginning to be engaged in tumor associated antigen peptide specific CTL from 2005 is established, TCR correlative study (Int J Hematol.2011,93:176-185) and tcr gene modification T cell research, respectively successfully by tumor associated antigen WT1 and Aurora kinase A specificity TCR transduction circulation CD8+T cell (Blood, 2011,118:1495-1503；Blood, 2012,119:368-376) TCR-T clone, is obtained, through external, internal identification, it was confirmed that its anti-lung cancer or leukaemia cell's Validity.
Tcr gene modification T cell technology increasingly show its important value in immunotherapy of tumors and it is fine before Scape, and the key precondition for generating TCR-T is to obtain the TCR with specific recognition tumour antigen, establishes tumour by external evoked Associated antigen polypeptide specific CTL is an important channel for realizing this target.
XAGE-1 (X antigen family member 1) be Cancer-testis antigen (Cancer-Testis antigen, CTA) one of family member, CTA are also known as tumour-hair tonic system (Tumor-Germline, TG) antigen.Such antigen, is only expressed in Testicle spermatogonia, without being expressed in normal tissue cell, meanwhile, research finds that CTA is expressed in a variety of histological types Tumour, thus also known as tomour specific shares antigen (tumor-specific shared antigens, TSSA).Due to testis Spermatogonium does not express major histocompatibility antigen compound (major histocompatibility complex, MHC) I Class molecule, therefore the immune response of the CTL of CTA induction is to testicular cell fanout free region, killing tumor cell.Exactly because this Characteristic makes ideal antigen of the CTA as immunotherapy of tumors.Mankind XAGE-1 belongs to GAGE/PAGE family, XAGE-1 gene It is positioned at Xp11.21-Xp11.22, including tetra- kinds of spliceosomes of XAGE-1a, XAGE-1b, XAGE-1c, XAGE-1d, XAGE-1b (X Antigen family, member 1b) full length gene 622bp, encode the amyloid protein precursor being made of 81 amino acid.XAGE-1b Kinds of tumors include breast cancer, prostate cancer, various types of lung cancer (gland cancer, squamous carcinoma and Small Cell Lung Cancer etc.), oophoroma, There is expression in melanoma, glioblastoma, lymthoma and leukaemia etc..The gene has XAGE1a, XAGE-1b, XAGE- 1c and XAGE-1d4 kind transcribes isomers, and wherein XAGE-1b is the dominant antigen that can be identified by immunocyte, and immunogenicity is very By force, high expression in adenocarcinoma of lung is especially in NSCLC, it has also become a novel targets of lung cancer immunization therapy.However, at present not yet There is the report that XAGE-1b Antigenic Peptide specific CTL has been established.Although some CTA polypeptid specificity CTL have built up and pass through mirror It is fixed, such as: NY-ESO-1 (New York esophageal squamous cell carcinoma 1) Peptide-specific CTL, MAGE antigen (melanoma-associated antigens), MAGE-1 specific CTL etc..But due to tumour it is heterogeneous with And the individual difference between tumor patient, obtaining enough tumor antigen peptide specific CTL clones is very important.Generally recognize Shorter for the length of polypeptide, the specific immunity ability that can induce is weaker, or even can not induce with therapeutic value completely Ctl clone；Conversely, polypeptide length is longer, specific CTL clone more can induce out.However the length of polypeptide is longer, synthesis is difficult Spend bigger, be more difficult to obtain pure polypeptide, the presence of impurity peptide on the formation of clone there are unpredictable influence, and polypeptide Purifying cost is high but is still difficult to avoid that the presence of impurity peptide, and the cost of this scheme is extremely high.Accordingly it is desirable to Shorten the length of polypeptide in the case where retaining polypeptide antigen as far as possible.It is attempted in technical solution disclosed in CN102428102A A part in XAGE-1b antigen is intercepted to induce humoral immunity or cellular immunity to tumour, but its result can not enable People is satisfied, and the length of polypeptide is not shorter than 16AA still.Small peptide the problem of there is likely to be poor specificities, how to obtain specificity Good and with good immunogenicity small peptide is an extremely challenging job.
CN106243213A, CN106279391A and CN106279392A disclose a series of XAGE-1b small peptides, illustrate short Peptide is feasible for inducing producing specificity immune response, but experimental data shows that it needs further to be improved.
Summary of the invention
The purpose of the present invention is to provide a kind of XAGE-1b nonapeptide and its applications.
The technical solution used in the present invention is:
XAGE-1b small peptide, sequence GSRQKKIRI.
XAGE-1b small peptide prepares the application in tumor-specific cytotoxicity T cell clone in induction, wherein XAGE-1b Short peptide sequence is GSRQKKIRI.
The abductive approach of tumor-specific cytotoxicity T cell, sequence are the XAGE-1b small peptide of GSRQKKIRI through dendron Shape cell offers to co-culture with CD8+T cell, and induction screening obtains tumor-specific cytotoxicity T cell.
A kind of tumour polypeptide vaccine is made of active antigens ingredient and adjuvant, and active antigens ingredient is that sequence is The XAGE-1b small peptide of GSRQKKIRI.
A kind of DC vaccine for oncotherapy is mainly thin for the XAGE-1b small peptide and dendron shape of GSRQKKIRI by sequence Born of the same parents load to obtain.
XAGE-1b small peptide is preparing the application in Diagnosis of Non-Small Cell Lung kit, wherein the sequence of XAGE-1b small peptide It is classified as GSRQKKIRI.
The beneficial effects of the present invention are:
The CTL that XAGE-1b antigen inducing peptide of the invention is established will not generate immune response to testicular cell, only kill swollen Oncocyte.Induce obtained ctl clone that there is good tumor cell specific lethal effect.Simultaneously during establishing CTL, The XAGE-1b Antigenic Peptide that filters out of inventor's discovery has and HLA affinity appropriate on DC cell and can effectively stimulate, lures Specific CTL s is given birth in artificial delivery, illustrates that it has the potentiality of good polypeptide vaccine and DC vaccine.By cloning XAGE-1b Antigenic Peptide Specificity TCR gene constructs viral vectors, and peripheral circulation CD8+T cell of transduceing obtains the T cell (TCR- of tcr gene modification T it) clones, equally there is tumor cell specific lethal effect similar with parental generation CTL, prompt it with good clinical conversion And actual application prospect.
Detailed description of the invention
Fig. 1 is expression of the XAGE-1b in lung adenocarcinoma cell；
Fig. 2 is the representative result of XAGE-1b (19-27) peptide specific CTL cell toxicity test, E: effector cell (Effectcell), T: target cell (Target cell), HLA-A*24 positive human T2 cell；
Fig. 3 be XAGE-1b (19-27) specific CTL HLA I class inhibit test (target cell: T2 cell, E/T=10: 1W6/32: anti-HLA I class molecule neutralizing antibody, the anti-HLA II class molecule neutralizing antibody of L243)；
Fig. 4 is XAGE-1b (31-39) specific CTL IFN-γ release test result；
Fig. 5 is XAGE-1b (55-63) specific CTL IFN-γ release test result.
Specific embodiment
XAGE-1b small peptide, sequence GSRQKKIRI.
XAGE-1b small peptide prepares the application in tumor-specific cytotoxicity T cell clone in induction, wherein XAGE-1b Short peptide sequence is GSRQKKIRI.
The abductive approach of tumor-specific cytotoxicity T cell, sequence are the XAGE-1b small peptide of GSRQKKIRI through dendron Shape cell offers to co-culture with CD8+T cell, and induction screening obtains tumor-specific cytotoxicity T cell.
A kind of tumour polypeptide vaccine is made of active antigens ingredient and adjuvant, and active antigens ingredient is that sequence is The XAGE-1b small peptide of GSRQKKIRI.
A kind of DC vaccine for oncotherapy is mainly thin for the XAGE-1b small peptide and dendron shape of GSRQKKIRI by sequence Born of the same parents load to obtain.
XAGE-1b small peptide is preparing the application in Diagnosis of Non-Small Cell Lung kit, wherein the sequence of XAGE-1b small peptide It is classified as GSRQKKIRI.
Tumour antigen selects XAGE-1b antigen, and XAGE-1b gene is located at X chromosome (Xp11.21-Xp11.22), overall length 622bp encodes the amyloid protein precursor being made of 81 amino acid.Amino acid sequence is following (to be originated from GenBank:NM_ 001097594.2):
MESPKKKNQQLKVGILHLGSRQKKIRIQLRSQCATWKVICKSCISQTPGINLDLGSGVKVKIIPKEEH CKMPEAGEEQPQV (SEQ ID NO:1).
Below with reference to experiment, technical solution of the present invention is further illustrated.Expression of the XAGE-1b in non-small cell lung cancer XAGE-1 is important member in CTA family, in addition to having wide expression in non-small cell lung cancer, in Several Kinds of Malignancy There is expression, establishes XAGE-1b Antigenic Peptide specific CTL clone and be extremely important.Inventor is random in early-stage study Choose surgical resection lung cancer specimen (make a definite diagnosis through pathologic finding and obtain patient's agreement), conventional line RT-PCR detection Expression of the XAGE-1b mRNA in NSCLC.
As a result as Fig. 1, display XAGE-1b have wide expression in lung adenocarcinoma cell.Inventor by have by oneself method, Comprehensive score is carried out to the CTL epitope of XAGE-1b antigen, with two different database (US National Institutes of Health Research Institutes In BIMAS, http://www-bimas.cit.nih.gov/molbio/hla_bind/ and Heidelberg, Germany biomedical information The heart SYFPEITHI, http://www.syfpeithi.de/) scoring product be the prediction epitope overall score, according to general comment Divide the epitope for predicting its CTL.Gained candidate peptide is synthesized by specialized company.
Based on prediction result, inventor randomly chooses 3 progress result verification therein, and specific experiment is as follows:
XAGE-1b (19-27) peptide specific ctl clone killing experiments:
Inventor establishes XAGE-1b (19-27) (GSRQKKIRI, (SEQ ID NO:2)) specific CTL clone, operation It is as follows:
The 10 of same health donor5A CD8+T cell passes through the 10 of load XAGE-1b (10-18) peptide4Between a Mo-DCs After stimulation in 1 week 2 times, then pass through self 105The PBMC of processed load XAGE-1b (19-27) peptide of isotope is stimulated 1 time Afterwards, it is obtained through standard cell poison experiment sieving.
T2 cell loads 5uM XAGE-1b (19-27) peptide as target cell, XAGE-1b (19-27) peptide specific of CTL Cytotoxicity is confirmed by LDH release test.
Experimental result such as Fig. 2, cytotoxicity test result confirm XAGE-1b (19-27) peptide specific CTL to HLA-A*24 sun Property target cell load polypeptide after there is good fragmentation effect, but to the target cell of unsupported polypeptide without obvious cytotoxicity, The preliminary proof CTL is XAGE-1b (19-27) peptide specific.
XAGE-1b (19-27) specific CTL HLA class I inhibits test
It whether is that HLA-Class I is restricted for clear XAGE-1b (19-27) specific CTL, inventor passes through thin in target Anti- HLA-Class I neutralizing antibody is added in born of the same parents HLA-A24+T2, repeats above-mentioned Antigenic Peptide Cytotoxicity assays, as a result such as Fig. 3, test result tentatively illustrate that the cytotoxicity of XAGE-1b (19-27) specific CTL is that HLA I class molecule is restrictive.
Based on above research, loading XAGE-1b antigen polypeptide by external DCs stimulates CD8+T cell in periphery that can build Vertical XAGE-1b antigen polypeptide specific CTL clone, has been established XAGE-1b (19-27) peptide specific ctl clone at present, preliminary logical Crossing CTL cytotoxicity test confirms its antigen peptide specific responsing reaction.
Using the above external evoked method for establishing XAGE-1b small peptide specific CTL clone, inventor has also set up HLA- The restricted XAGE-1b of A*24 (31-39) (SQCATWKVI (SEQ ID NO:3)) and XAGE-1b (55-63) (GSGVKVKII (SEQ ID NO:4)) specific CTL clone, its polypeptid specificity immune response effect is confirmed (such as by IFN-γ release test Fig. 4,5).
Above-mentioned experimental data shows that the CTL epitope that inventor establishes is extremely effective, prediction result and experimental result symbol Conjunction property is very good.As it can be seen that by the way that above-mentioned XAGE-1b small peptide GSRQKKIRI is offered and cytotoxicity lymph T through Dendritic Cells Cell co-cultures, and inducible screening obtains specific for tumour antigen cytotoxic T lymphocyte.This specific for tumour antigen is thin Cytotoxic T Lymphocytes can be used for the treatment of tumour.
Above-mentioned XAGE-1b small peptide GSRQKKIRI and Dendritic Cells (dendritic cell, DC) load are fed back, it can To be used for tumour immunity as DC vaccine, stimulation body generates the antitumor T cell of polypeptid specificity, and then realizes controlling for tumour It treats.
Above-mentioned XAGE-1b small peptide GSRQKKIRI can be used for measuring the antibody level that polypeptide is directed in subject sample, into And it is used for the diagnosis of non-small cell lung cancer.
XAGE-1b small peptide length of the invention is only 9 amino acid, and chemical synthesis difficulty is small, can directly synthesize to obtain High-purity product, application cost substantially reduce, while definite effect, have good application potential.
SEQUENCE LISTING
<110>Guangzhou Mei Sa Biotechnology Co., Ltd
<120>a kind of XAGE-1b nonapeptide and its application
<130> XAGE-1b(19-27)
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 81
<212> PRT
<213>mankind
<400> 1
Met Glu Ser Pro Lys Lys Lys Asn Gln Gln Leu Lys Val Gly Ile Leu
1 5 10 15
His Leu Gly Ser Arg Gln Lys Lys Ile Arg Ile Gln Leu Arg Ser Gln
20 25 30
Cys Ala Thr Trp Lys Val Ile Cys Lys Ser Cys Ile Ser Gln Thr Pro
35 40 45
Gly Ile Asn Leu Asp Leu Gly Ser Gly Val Lys Val Lys Ile Ile Pro
50 55 60
Lys Glu Glu His Cys Lys Met Pro Glu Ala Gly Glu Glu Gln Pro Gln
65 70 75 80
Val
<210> 2
<211> 9
<212> PRT
<213>artificial small peptide
<400> 2
Gly Ser Arg Gln Lys Lys Ile Arg Ile
1 5
<210> 3
<211> 9
<212> PRT
<213>artificial small peptide
<400> 3
Ser Gln Cys Ala Thr Trp Lys Val Ile
1 5
<210> 4
<211> 9
<212> PRT
<213>artificial small peptide
<400> 4
Gly Ser Gly Val Lys Val Lys Ile Ile
1 5

Claims (6)

1.XAGE-1b small peptide, sequence GSRQKKIRI.
2.XAGE-1b small peptide prepares the application in tumor-specific cytotoxicity T cell clone in induction, wherein XAGE-1b is short Peptide is as described in claim 1.
3. the abductive approach of tumor-specific cytotoxicity T cell, it is characterised in that: use XAGE-1b described in claim 1 Small peptide offers to co-culture with CD8+T cell through Dendritic Cells, and induction screening obtains tumor-specific cytotoxicity T cell.
4. a kind of tumour polypeptide vaccine is made of active antigens ingredient and adjuvant, it is characterised in that: active antigens ingredient is as weighed Benefit requires the 1 XAGE-1b small peptide.
5. a kind of DC vaccine for oncotherapy, is mainly added by XAGE-1b small peptide described in claim 1 and Dendritic Cells Load obtains.
6.XAGE-1b small peptide is preparing the application in Diagnosis of Non-Small Cell Lung kit, wherein XAGE-1b small peptide such as right It is required that described in 1.
CN201910233750.6A 2018-12-19 2019-03-26 A kind of XAGE-1b nonapeptide and its application Pending CN110016073A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201811555751 2018-12-19
CN2018115557514 2018-12-19

Publications (1)

Publication Number Publication Date
CN110016073A true CN110016073A (en) 2019-07-16

Family

ID=67189966

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910233750.6A Pending CN110016073A (en) 2018-12-19 2019-03-26 A kind of XAGE-1b nonapeptide and its application

Country Status (1)

Country Link
CN (1) CN110016073A (en)

Similar Documents

Publication Publication Date Title
CN109970846A (en) A kind of tumor associated antigen XAGE-1b nonapeptide and its application
CN106243213B (en) A kind of tumor associated antigen XAGE-1b small peptides and application
CN106279392B (en) Tumor associated antigen XAGE-1b small peptides and its application
JP4185147B2 (en) KDR peptide and vaccine containing the same
FI121710B (en) Melanoma antigens
JP5292550B2 (en) T cell receptor β chain gene and α chain gene
CN103570818B (en) Tumor antigenic polypeptide and the purposes as tumor vaccine thereof
Blanchard et al. Vaccines against advanced melanoma
Saif et al. Novel prostate acid phosphatase‐based peptide vaccination strategy induces antigen‐specific T‐cell responses and limits tumour growth in mice
KR20060129393A (en) Epitope/peptide recognized by hla-a2402-restricted ep-cam-specific ctl and use of the same
US10765729B2 (en) Tumor antigen peptide
JP6960179B2 (en) Tumor antigen peptide
EP2270144B1 (en) Partial peptide of survivin presented on mhc class ii molecule and use thereof
JP7266834B2 (en) tumor antigen peptide
WO2010107116A1 (en) Hla-a24-binding cancer antigen peptide derived from sox2
CN106279391B (en) Tumor associated antigen XAGE-1b small peptide and application
US9260499B2 (en) Tumor antigen peptide and use thereof
CN108138170A (en) Peptide derived from DEPDC1 and the vaccine for including them
CN110016073A (en) A kind of XAGE-1b nonapeptide and its application
CN106715461A (en) KOC1-dervied peptide and vaccine including same
Nagarajan et al. Helicase antigen (HAGE)‐derived vaccines induce immunity to HAGE and ImmunoBody®‐HAGE DNA vaccine delays the growth and metastasis of HAGE‐expressing tumors in vivo
TW201100090A (en) C6orf167 peptides and vaccines containing the same
CN107074908A (en) Peptide derived from CDCA1 and the vaccine containing them
JP5424640B2 (en) Target-specific identification method for immunogenic peptides
JP2002360251A (en) Hla-a24-restricted tumor antigen peptide derived from mn/ca9

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20190716

WD01 Invention patent application deemed withdrawn after publication