Tumor antigen peptides, antibodies, detection kits, pharmaceutical compositions or vaccines and uses
By providing tumor-specific antigen short peptides and their derivatives derived from the DHRS4 gene mutation site, combined with recombinant nucleic acid molecules and engineered cells, the problem of insufficient accuracy in neoantigen recognition has been solved, achieving highly efficient immunotherapy for lung adenocarcinoma.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HARBIN MEDICAL UNIVERSITY
- Filing Date
- 2026-02-06
- Publication Date
- 2026-06-05
AI Technical Summary
Existing technologies lack sufficient accuracy in recognizing neoantigen short peptides and lack immunogenicity verification, resulting in limited immunotherapy response rates. In particular, it is difficult to obtain a sufficient number and quality of effective targets for cold tumors with low mutation burden.
It provides tumor-specific antigen short peptides derived from DHRS4 gene mutation sites, which are modified to form multi-epitope peptides and delivered and stimulated with recombinant nucleic acid molecules, lipid nanoparticle formulations, engineered cells and specific antibodies.
These short peptides can be efficiently presented, inducing specific CD8+ T cell immune responses and exhibiting significant cytotoxicity against tumor cells expressing corresponding mutations, providing a material basis for precision immunotherapy of lung adenocarcinoma.
Smart Images
Figure CN122146636A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biotechnology and provides a tumor antigen peptide, antibody, detection kit, pharmaceutical composition or vaccine and its uses. Background Technology
[0002] Lung adenocarcinoma is one of the most common and deadliest malignant tumors worldwide, accounting for approximately 40%–50% of non-small cell lung cancer (NSCLC). Although surgery, radiotherapy, chemotherapy, and targeted therapy have shown some efficacy in some patients, the overall prognosis remains unsatisfactory, especially after the development of drug resistance, metastasis, or recurrence, where the effectiveness of traditional treatments significantly declines. Recent studies have shown that the occurrence of lung adenocarcinoma is closely related to immune escape. Tumor cells evade the surveillance and clearance of the body's immune system through mechanisms such as downregulating antigen-presenting molecules, secreting immunosuppressive factors, and inducing immune tolerance. Therefore, restoring or enhancing the body's specific immune recognition ability against tumors has become an important strategy for improving the treatment efficacy of lung adenocarcinoma. Personalized immunotherapy targeting neoantigens has emerged in this context, providing a new direction for the precision treatment of lung adenocarcinoma. Tumor immunotherapy, by activating the patient's own immune system to specifically recognize and eliminate tumor cells, is a crucial direction in current cancer treatment. Tumor-specific mutant antigens (neoantigens), originating from unique gene mutations in tumor cells, are expressed and presented only in tumor tissue, effectively avoiding damage to normal tissues and exhibiting high specificity and safety. After intracellular processing, neoantigens are presented to the cell surface by major histocompatibility complex (MHC) or human leukocyte antigen (HLA) molecules, thereby being recognized by cytotoxic T cells (CTLs) and inducing a specific immune response. Therefore, obtaining short peptides of antigens that can be efficiently presented and elicit a strong immune response is a key foundation for the development of precision immunotherapy drugs.
[0003] Currently, neoantigen discovery relies heavily on sequencing prediction or immunopeptidomics identification, but both approaches have limitations: the former has a high false-positive rate, making it difficult to accurately reflect actual presentation and immunogenicity in vivo; the latter, while directly validating presented peptides, is limited by detection sensitivity and sample requirements, making it difficult to comprehensively cover low-abundance but crucial neoantigens. Particularly for "cold tumors" with low mutational burdens, traditional methods often fail to obtain a sufficient number and quality of effective targets, resulting in limited immunotherapy response rates and unmet clinical needs. Summary of the Invention
[0004] To address the shortcomings of existing technologies, such as insufficient accuracy in recognizing neoantigen short peptides and a lack of immunogenicity verification, this invention provides a set of tumor-specific antigenic short peptides derived from DHRS4 gene mutation sites. Experimental results show that these short peptides can be efficiently presented by human HLA molecules and can effectively induce specific CD8+ expression. +T-cell immune responses and significant cytotoxic effects on tumor cells expressing corresponding mutations.
[0005] This invention is achieved through the following technical solution: The primary objective of this invention is to provide a tumor antigen peptide and its derivatives, wherein the amino acid sequence of the tumor antigen peptide is shown in SEQ ID NO:1.
[0006] In one embodiment of the present invention, the antigenic peptide comprises a mutant epitope sequence derived from the DHRS4 protein.
[0007] As one embodiment of the present invention, the tumor antigen peptide is a functional derivative thereof, having the core epitope of SEQ ID NO:1 and comprising one or more of the following modifications: 1) N-terminal or C-terminal modification; 2) amino acid substitution or length variation; 3) linkage with other peptides to form a multi-epitope peptide; 4) cyclization, PEGylation, fatty acidation or linker peptide modification.
[0008] The second objective of this invention is to provide a delivery system comprising recombinant nucleic acid molecules, expression vectors, or lipid nanoparticle formulations.
[0009] As one embodiment of the present invention, the recombinant nucleic acid molecule encodes any of the aforementioned tumor antigen peptides, or encodes a functional derivative having at least 90% sequence homology.
[0010] In one embodiment of the present invention, the expression vector comprises the aforementioned recombinant nucleic acid molecule.
[0011] As one embodiment of the present invention, the lipid nanoparticle formulation encapsulates the aforementioned recombinant nucleic acid molecules and is suitable for in vivo delivery and inducing antigen-specific immune responses.
[0012] The third objective of this invention is to provide an engineered cell that contains the aforementioned recombinant nucleic acid molecule or expression vector, or presents the aforementioned antigenic peptide on its cell surface.
[0013] As one embodiment of the present invention, the cell is selected from any of the following: 1) Host cell: a host cell transfected with an expression vector containing the aforementioned recombinant nucleic acid molecule, wherein the host cell is selected from prokaryotic cells, eukaryotic cells or tool cell lines; 2) Engineered tool cell: dendritic cells, antigen-presenting cells or genetically modified cell lines that present the aforementioned tumor antigen peptide on their surface; 3) TCR-T cell: a cell that expresses a TCR-T cell receptor that can specifically recognize the aforementioned tumor antigen peptide and HLA molecule complex.
[0014] The fourth objective of this invention is to provide a specific antibody that can specifically bind to any of the aforementioned tumor antigen peptides.
[0015] As one embodiment of the present invention, the antibody is a monoclonal antibody, a polyclonal antibody, a humanized antibody, or a single-chain variable region antibody (scFv).
[0016] The fifth objective of this invention is to provide a tumor detection kit comprising any of the aforementioned antigenic peptides or any of the aforementioned recombinant nucleic acid molecules.
[0017] As one embodiment of the present invention, it further includes: a specific antibody against the antigenic peptide, and / or a primer pair for detecting the nucleic acid molecule.
[0018] The sixth objective of this invention is to provide in vitro applications of the aforementioned tumor antigen peptides or recombinant nucleic acid molecules, selected from in vitro detection or in vitro cell preparation.
[0019] As one embodiment of the present invention, 1) in vitro detection: using any of the aforementioned tumor antigen peptides to detect the presence of specific T cells in the sample; or using any of the aforementioned recombinant nucleic acid molecules to detect DHRS4 mutation status; 2) in vitro preparation: using any of the aforementioned tumor antigen peptides and / or recombinant nucleic acid molecules to prepare and / or isolate effector T cells in vitro.
[0020] The seventh objective of this invention is to provide a pharmaceutical composition or vaccine comprising any of the aforementioned tumor antigen peptides or antibodies, and a pharmaceutically acceptable carrier or excipient. The carrier or excipient includes, but is not limited to, one or more of the following: immune adjuvants, diluents, binders, disintegrants, lubricants, coating materials, solvents, surfactants, stabilizers, preservatives, flavoring agents, or coloring agents.
[0021] As one embodiment of the present invention, the composition is prepared as an injection or cell infusion dosage form.
[0022] The eighth objective of this invention is to provide the use of any of the aforementioned tumor antigen peptides or any of the aforementioned antibodies in the preparation of a drug for treating tumors.
[0023] In one embodiment of the present invention, the tumor is lung adenocarcinoma.
[0024] The beneficial effects of this invention are as follows: the antigenic peptide of this invention (especially the sequence containing the DHRS4 mutant epitope) exhibits good immunogenicity and antitumor activity in in vitro models, and can serve as a key active component in the preparation of novel tumor vaccines, TCR-T cell therapy, or immune adjuvants. Particularly in the treatment research of lung adenocarcinoma, this antigenic peptide and its related preparations (such as LNPs and antibody drugs) have shown significant specificity and efficacy, providing an important material basis and technical support for the development of precision immunotherapy drugs targeting solid tumors. Attached Figure Description
[0025] Figure 1 Mass spectrometry analysis chromatogram; Figure 2 HLA-A*11:01 genotyping waste blood samples were subjected to antigen loading using negative control peptides and antigenic peptides, respectively. Figure 3 Figure 1. Flow cytometry results of T cell activation by antigen peptides of this invention; Figure 4 ELISA results of T cells stimulating INF-γ secretion by antigen peptides of this invention; Figure 5 The results of the immunofluorescence experiment showing how the antigen peptide of this invention promotes T cell killing of tumors are shown in the figure. Detailed Implementation
[0026] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise defined, the technical terms used in the following embodiments have the same meaning as commonly understood by those skilled in the art to which this invention pertains. Unless otherwise specified, the experimental reagents used in the following embodiments are all conventional biochemical reagents; the experimental methods, unless otherwise specified, are all conventional methods.
[0027] Unless otherwise specified, the experimental methods in the following embodiments are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available. The present application will now be described in detail with reference to specific embodiments.
[0028] Example 1 I. Establishment, Synthesis, and Identification of Short Antitumor Peptides 1.1 Antigenic peptide prediction Lung cancer tumor data, including RNA-seq and mass spectrometry data, were screened using the public database GEO. Mutation frequencies and related short peptides were analyzed. The HLA binding affinity, antigen peptide-MHC complex stability, and antigen peptide processing and presentation efficiency of the relevant antitumor short peptides were predicted using NetMHCpan HLA-I / II, NetMHCstapan, and NetCTLpan, respectively. The peptides were then weighted and ranked based on mutation frequency, affinity, stability, and processing and presentation efficiency scores to identify antigen peptides derived from DHRS4 mutations. The mutations were selected from at least one of missense, nonsense, or frameshift mutations.
[0029] The sequences used in this application are as follows: SEQ ID NO:1 AGCSGWTRK. 1.2 Antigenic peptide synthesis The aforementioned antigenic peptides were synthesized using a solid-phase synthesis method.
[0030] 1) The solid support resin is swollen with an organic solvent. Add a sufficient amount of polar organic solvent and shake to swell for 30 minutes at room temperature. 2) Fmoc Deprotection: Remove the solvent from the reactor, add a DMF solution containing 15% piperidine (v / v), and shake the reaction for 15 minutes to remove the Fmoc protecting group at the N-terminus of the peptide chain, exposing the free amino group; this step is usually repeated once to ensure complete removal. After completion, wash the resin 5 times with DMF to thoroughly remove the deprotecting agent and byproducts; 3) Amino acid coupling: An activation solution is prepared by dissolving 3 equivalents of Fmoc-protected amino acid, an equimolar amount of condensing agent HBTU or HATU, and 6 equivalents of organic base DIPEA or NMM in an appropriate amount of DMF, and immediately added to the reactor. The reaction is carried out with shaking at room temperature for 90 minutes to allow the activated amino acid to form peptide bonds with the free amino groups on the resin. After the reaction is complete, the resin is thoroughly washed with DMF. 4) Final cleavage and deprotection: After the target sequence is assembled, the resin is treated with a cleavage mixture consisting of trifluoroacetic acid, water and a scavenging agent to cleave the peptide chain from the resin and remove all side chain protecting groups at the same time to obtain crude peptide. 5) Purification: The crude peptide solution was initially purified by precipitation, and then further purified by preparative high performance liquid chromatography. After freeze-drying, the final antigen peptide was obtained, which has the following sequence: AGCSGWTRK.
[0031] 1.3 Mass spectrometry identification The short peptide sample was dissolved and diluted with an acetonitrile-water solution containing 0.1% formic acid. Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALS) was used for analysis. The sample solution was injected into the mass spectrometer, and data was acquired in positive ion mode to obtain the molecular ion peak information of the sample, such as... Figure 1 As shown. By comparing the mass spectrometry results with the theoretical values, the theoretical molecular weight of the synthesized short peptide AGCSGWTRK was 965.09 Da, and the measured molecular weight was 965.73 Da. The two are basically consistent, confirming that the sample synthesis was successful.
[0032] II. Immunogenicity Identification of Tumor Antigen Peptides 1.1 Antigen Presentation HLA-A*11:01 genotyping waste blood samples were collected, and human peripheral blood mononuclear cells (PBMCs) were separated by density gradient centrifugation. The PBMCs were antigen-loaded with negative control peptide and the aforementioned SEQ ID NO:1 antigen peptide at a concentration of 10 ng / mL. Each group was co-stimulated with CD3. The cells were incubated for 6 hours in a 5% CO2, 37°C incubator supplemented with 1% penicillin-streptomycin, 10% fetal bovine serum and IL-2 in complete culture medium 1640.
[0033] 1.2 Tetramer Binding Ability Test 1) Dissolve the antigenic peptide to 500 μM using PBS, add 20 μL of the diluted peptide solution and 20 μL of HLA-A*11:01 Flex-T to a 96-well plate. TM The monomer UVX (200 μg / mL) was mixed by pipetting and centrifugation at 2500g for 2 minutes at 4°C, followed by irradiation with a UV crosslinker for 30 minutes to perform peptide exchange.
[0034] 2) Transfer 30 μL of peptide-exchanged monomer to a 1.5 ml centrifuge tube, add 3.3 μL of streptavidin, mix well, and incubate on ice in the dark for 30 minutes.
[0035] 3) Prepare a blocking solution using 192.4 μL PBS, 1.6 μL 50 mM D-biotin and 6 μL 10% sodium azide. After mixing, add 2.4 μL of blocking solution to terminate the reaction.
[0036] 4) Place the tubes at 4°C and incubate overnight in the dark.
[0037] 5) Centrifuge the assembled tetramer at 4°C, 2500g for 5 minutes, and repeat the process 2 × 10⁻⁶ times. 6 Add 2 μL of tetramer to the cells, mix well, and incubate on ice in the dark for 30 minutes.
[0038] 6) Wash the cells twice with flow cytometry buffer and then resuspend them in flow cytometry buffer for detection.
[0039] The results are as follows Figure 2 As shown, the antigenic peptide can be specifically recognized by the HLA-A*11:01 tetramer.
[0040] 1.3 Detection of activation markers Flow cytometry was used to label target cells with FITC-CD3, APC-CD8, and PE-CD69. CD69 is a protein expressed on the surface of immune cells, belonging to the C-type lectin receptor family, and can act as a CD8 receptor. + Markers of early T cell activation.
[0041] 1) Collect PBMCs after antigen presentation (2.1) and adjust the cell count to 1×10⁻⁶.6 Centrifuge at 1000 rpm for 5 minutes.
[0042] 2) Discard the supernatant, resuspend the cells in 500 μL of flow cytometry buffer, wash the culture medium, and centrifuge at 1000 rpm for 5 minutes.
[0043] 3) Discard the supernatant, resuspend the cells in 100 μL of flow cytometry buffer, add 5 μL each of FITC-CD3, APC-CD8, and PE-CD69 flow cytometry antibodies to each sample, and incubate at 4°C in the dark for 15 minutes.
[0044] 4) After incubation, add 500 μL of flow cytometry buffer to wash and centrifuge at 100 rpm for 5 minutes.
[0045] 5) Resuspend the cells in 500 μL of flow cytometry buffer and perform the analysis using a laminar flow cytometer.
[0046] The results are as follows Figure 3 As shown, the results indicate that CD3+ is generated after loading the antigenic peptide of SEQ ID NO:1. + CD8 + CD69 in double-positive cell population + The proportion of cells increased significantly. This result indicates that the antigenic peptide of SEQ ID NO:1 can significantly stimulate CD3 in HLA-A*11:01 genotyped PBMCs. + CD8 + T cell activation.
[0047] 1.4 Cytokine Detection CD8+ after antigen peptide stimulation was detected by ELISA. + The secretion of IFN-γ and TNF-α in the supernatant of T cell culture was detected.
[0048] 1) Equilibrate the kit to room temperature and dilute the concentrated washing solution with double-distilled water at a ratio of 24:1.
[0049] 2) Dilute the standard according to the ratio to prepare 1000, 500, 250, 125, 62.5, 31.25, 15.63, and 0 pg / mL.
[0050] 3) Add 100 μL of the sample and standard to each of the 96-well microplates pre-coated with antibody and incubate at 37°C for 90 minutes.
[0051] 4) Discard the liquid in the wells, add 100 μL of biotinylated antibody working solution to each well, cover the plate with a membrane, and incubate at 37°C for 1 hour.
[0052] 5) Shake off the liquid in the wells and soak each well in washing solution for 1 minute. Repeat 3 times and shake off the liquid. If there is no time, add 100 μL of HRP enzyme conjugate working solution, cover the microplate with a membrane, and incubate at 37°C for 30 minutes.
[0053] 6) Discard the liquid in the wells, wash the plate 5 times, add 90 μL of substrate solution to each well, cover the plate with a membrane, and incubate at 37°C in the dark for 15 minutes.
[0054] 7. Add 50 μL of stop solution to each well to terminate the reaction and use a microplate reader to detect the optical density (OD value) of each well at a wavelength of 450 nm.
[0055] 8) Calculate the standard curve using the standard and calculate the content of IFN-γ and TNF-α in the supernatant.
[0056] The results are as follows Figure 4 As shown, the results indicated that the levels of IFN-γ and TNF-α in the cell supernatant after loading the antigenic peptide of SEQ ID NO:1 were significantly increased.
[0057] 1.5 Cytotoxicity Detection 1) T cells loaded with antigen were co-cultured with target cells (normal lung epithelial cell line Beas-2B and lung adenocarcinoma cell line A549) at a ratio of 10:1. Cell viability was detected using a live-dead cell staining kit after 3 days.
[0058] 2) Remove T cells and wash adherent cells with PBS, then remove the PBS.
[0059] 3) Add 200 μL of staining buffer and 2 μL of Calcein AM Solution to each well, and incubate at 37°C for 20 minutes.
[0060] 4) Add 2 μL of PI Solution to each well. 5) After incubation, use a fluorescence microscope to count the number of live and dead cells.
[0061] The results are as follows Figure 5 As shown, the results indicated that antigen-loaded T cells significantly killed A548 lung adenocarcinoma cells, but had no significant toxicity to normal lung epithelial cells.
[0062] Furthermore, the tumor antigen peptide described in this application may also be a functional derivative thereof, having the core epitope of SEQ ID NO:1 and comprising one or more of the following modifications: N-terminal or C-terminal modification; Amino acid substitutions or length variations; It can link with other peptides to form multi-epitope peptides; Cyclation, PEGylation, fatty acidation, or adaptor peptide modification.
[0063] Further research provides a recombinant nucleic acid molecule encoding the tumor antigen peptide mentioned in this application, wherein the recombinant nucleic acid molecule has a functional variant with at least 90% sequence homology. The functional variant retains the biological activity of the encoded antigen peptide, such as soluble expression in prokaryotic / eukaryotic expression systems or improved translation efficiency.
[0064] Furthermore, an expression vector is provided, comprising the aforementioned recombinant nucleic acid molecule. The vector can be selected from conventional vector types in the field of gene delivery, such as plasmids.
[0065] An application as engineered cells is provided, comprising: 1) host cells transfected with the above-mentioned expression vector, wherein the host cells are selected from prokaryotic cells, eukaryotic cells, or tool cell lines; 2) engineered tool cells, wherein the cell surface presents any of the aforementioned tumor antigen peptides, wherein the cells are selected from dendritic cells, antigen-presenting cells, or genetically modified cell lines; and 3) TCR-T cells, wherein the expressed TCR-T cell receptor is capable of specifically recognizing any of the aforementioned tumor antigen peptides and HLA molecules complexes.
[0066] Based on providing a tumor antigen peptide, an antibody is provided that can specifically bind to the tumor antigen peptide. The antibody can be a monoclonal antibody, a polyclonal antibody, a humanized antibody, or a single-chain variable region antibody (scFv).
[0067] A tumor detection kit comprising a tumor antigen peptide or a recombinant nucleic acid molecule. It also includes: a specific antibody against the antigen peptide, and / or a primer pair for detecting the nucleic acid molecule.
[0068] An in vitro detection method is provided, which uses any of the aforementioned tumor antigen peptides to detect the presence of specific T cells in a sample, or uses any of the aforementioned nucleic acid molecules to detect DHRS4 mutation status.
[0069] A method for preparing effector T cells in vitro is provided, wherein effector T cells are prepared and / or isolated using any of the aforementioned tumor antigen peptides and / or the aforementioned recombinant nucleic acid molecules.
[0070] A tumor vaccine is provided, comprising any of the aforementioned tumor antigen peptides and an immune adjuvant.
[0071] A pharmaceutical composition is provided, comprising any of the aforementioned antigenic peptides and a pharmaceutically acceptable carrier or excipient; or any of the aforementioned antibodies and a pharmaceutically acceptable carrier or excipient. The pharmaceutically acceptable carrier or diluent, binder, disintegrant, lubricant, coating material, solvent, surfactant, stabilizer, preservative, flavoring agent, or colorant may be one or more of these. The pharmaceutical composition is an injectable or cell-infusion dosage form.
[0072] A lipid nanoparticle formulation for delivering any of the aforementioned recombinant nucleic acid molecules is provided, the formulation being suitable for in vivo delivery and inducing an antigen-specific immune response.
[0073] Provided for use of any of the aforementioned antigenic peptides or antibodies in the preparation of pharmaceutical formulations for the treatment of lung adenocarcinoma.
[0074] Unless otherwise specified, all of the above can be prepared or implemented using existing publicly available technologies or common knowledge, as well as methods commonly used by those skilled in the art.
[0075] The present invention has been described in detail above, but it should be understood that the above description does not constitute a limitation of the present invention. For those skilled in the art, the present invention can be implemented within a wide range under equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and various modifications, alterations, or improvements can be made to the present invention, including adjustments made using conventional techniques known in the art. Although the invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. A tumor antigen peptide, characterized in that, The amino acid sequence of the tumor antigen peptide is shown in SEQ ID NO:1; or, the tumor antigen peptide is a functional derivative of SEQ ID NO:1, the derivative having the core epitope of SEQ ID NO:1 and including one or more of the following modifications: 1) N-terminal or C-terminal modification; 2) amino acid substitution or length variation; 3) linkage with other peptides to form a multi-epitope peptide; 4) cyclization, PEGylation, fatty acidation or linker peptide modification.
2. The tumor antigen peptide according to claim 1, characterized in that, The tumor antigen peptide contains a mutant epitope sequence derived from the DHRS4 protein.
3. A recombinant nucleic acid molecule, characterized in that, It encodes the tumor antigen peptide as described in claim 1 or 2, or encodes a functional derivative having at least 90% sequence homology with the tumor antigen peptide.
4. A lipid nanoparticle formulation for delivering the recombinant nucleic acid molecule as described in claim 3, characterized in that, The formulation is suitable for in vivo delivery and inducing antigen-specific immune responses.
5. An engineered cell, characterized in that, The cells are selected from any of the following: 1) Host cells: transfected with an expression vector containing the recombinant nucleic acid molecule of claim 3, wherein the host cells are selected from prokaryotic cells, eukaryotic cells or tool cell lines; 2) Engineered tool cells: dendritic cells, antigen-presenting cells or genetically modified cell lines that present the tumor antigen peptide of claim 1 or 2 on their surface; 3) TCR-T cells: cells that express TCR-T cell receptors that specifically recognize the tumor antigen peptide and HLA molecule complex of claim 1 or 2.
6. A specific antibody, characterized in that, The antibody can specifically bind to the tumor antigen peptide described in claim 1 or 2.
7. A tumor detection kit, characterized in that, The kit contains the antigenic peptide as described in claim 1 or 2, or the recombinant nucleic acid molecule as described in claim 3; preferably, the kit further includes: 1) a specific antibody against the antigenic peptide; 2) a primer pair for detecting the nucleic acid molecule.
8. The in vitro application of the tumor antigen peptide of claim 1 or 2 or the recombinant nucleic acid molecule of claim 3, selected from any of the following uses: 1) In vitro detection: using the tumor antigen peptide of claim 1 or 2 to detect the presence of specific T cells in a sample, or using the recombinant nucleic acid molecule of claim 3 to detect DHRS4 mutation status; 2) In vitro preparation: using the tumor antigen peptide of claim 1 or 2 or / and the recombinant nucleic acid molecule of claim 3 to prepare or / and isolate effector T cells in vitro.
9. A pharmaceutical composition or vaccine, characterized in that, It contains an effective amount of the active ingredient and a pharmaceutically acceptable carrier or excipient; 1) the active ingredient is selected from: The tumor antigen peptide of claim 1 or 2 or the antibody of claim 6; 2) the carrier or excipient includes, but is not limited to, one or more of the following: immune adjuvant, diluent, binder, disintegrant, lubricant, coating material, solvent, surfactant, stabilizer, preservative, flavoring agent or coloring agent.
10. A pharmaceutical use, characterized in that, Use of the tumor antigen peptide of claim 1 or 2 or the antibody of claim 6 in the preparation of a tumor treatment drug or vaccine.