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Epitope polypeptide CD44-P2 based on prostatic cancer stem cell marker CD44 and application of epitope polypeptide CD44-P2

A CD44-P2, antigen epitope technology, applied in CD44, receptor/cell surface antigen/cell surface determinant, application, etc., can solve the problem of limited killing effect, low specificity of anti-tumor treatment technology, easy tumor recurrence, etc. problem, to reduce recurrence, enhance clear effect, and enhance suppression effect

Inactive Publication Date: 2019-04-12
深圳市龙华区人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing anti-tumor treatment techniques have low specificity, limited killing effect, and are prone to tumor recurrence

Method used

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  • Epitope polypeptide CD44-P2 based on prostatic cancer stem cell marker CD44 and application of epitope polypeptide CD44-P2
  • Epitope polypeptide CD44-P2 based on prostatic cancer stem cell marker CD44 and application of epitope polypeptide CD44-P2
  • Epitope polypeptide CD44-P2 based on prostatic cancer stem cell marker CD44 and application of epitope polypeptide CD44-P2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Design and synthesis of CD44 epitope short peptide

[0081] 1. Search the full amino acid sequence (NP_000601.3) of human CD44 protein from the NCBI Genbank, which contains 742 amino acids. Its amino acid sequence is as follows (SEQ ID NO: 3):

[0082] MDKFWWHAAWGLCLVPLSLAQIDLNITCRFAGVFHVEKNGRYSISRTEAADLCKAFNSTLPTMAQMEKALSIGFETCRYGFIEGHVVIPRIHPNSICAANNTGVYILTSNTSQYDTYCFNASAPPEEDCTSVTDLPNAFDGPITITIVNRDGTRYVQKGEYRTNPEDIYPSNPTDDDVSSGSSSERSSTSGGYIFYTFSTVHPIPDEDSPWITDSTDRIPATTLMSTSATATETATKRQETWDWFSWLFLPSESKNHLHTTTQMAGTSSNTISAGWEPNEENEDERDRHLSFSGSGIDDDEDFISSTISTTPRAFDHTKQNQDWTQWNPSHSNPEVLLQTTTRMTDVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTSTIQATPSSTTEETATQKEQWFGNRWHEGYRQTPKEDSHSTTGTAAASAHTSHPMQGRTTPSPEDSSWTDFFNPISHPMGRGHQAGRRMDMDSSHSITLQPTANPNTGLVEDLDRTGPLSMTTQQSNSQSFSTSHEGLEEDKDHPTTSTLTSSNRNDVTGGRRDPNHSEGSTTLLEGYTSHYPHTKESRTFIPVTSAKTGSFGVTAVTVGDSNSNVNRSLSGDQDTFHPSGGSHTTHGSESDGHSHGSQEGGANTTSGPIRTPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVINSGNGAVEDRKPSGLNGEASKSQEMVHLVNKESSETPDQFMTA...

Embodiment 2

[0090] The epitope polypeptide CD44-P2 of the CD44 protein epitope provided in Example 1 of the present invention was used to study the killing effect of CD44-induced effector T cells on the human prostate cancer cell LNCAP-multicellular sphere. It includes the following steps:

[0091] 1. Isolation and preparation of peripheral blood mononuclear cells: Collect 10 mL of peripheral blood from HLA-A2 positive healthy volunteers into two centrifuge tubes, centrifuge at 2000r / min for 5 minutes, discard the supernatant, mix the pelleted cells, and add physiological Saline to 25mL to fully suspend the precipitated cells to form a blood cell suspension. Take another centrifuge tube, add 20 mL of lymphocyte separation solution, and use a dropper to slowly transfer the blood cell suspension to the surface of the lymphocyte separation solution to form a clear interface between the two.

[0092] 2. After centrifuging the above-mentioned centrifuge tube with a clear interface at 2000r / min for...

Embodiment 3

[0106] The epitope polypeptide CD44-P2 of the CD44 protein epitope provided in Example 1 of the present invention was used to study the killing effect of CD44-induced effector T cells on human prostate cancer cell VCaP-multicellular spheres.

[0107] 1. The isolation and preparation method of peripheral blood mononuclear cells is the same as in Example 2.

[0108] 2. The method of culturing DC-CIK cells is the same as in Example 2.

[0109] 3. The preparation of the target cell VCaP-multicellular spheroid cells and the method for detecting the killing effect are the same as in Example 2.

[0110] Such as Figure 6A with Figure 6B As shown, the comparative analysis by flow cytometry showed that the proportion of CIK cells activated by the CD44 epitope polypeptide was significantly increased; at day 0 ( Figure 6A ), flow cytometry analysis results show that the proportion of CD3 positive T cells is 65.20%, the proportion of CD4 positive T cells is 35.50%, the proportion of CD8 positive...

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Abstract

The invention provides epitope polypeptide CD44-P2 based on a prostatic cancer stem cell marker CD44 and an application of the epitope polypeptide CD44-P2 and relates to the technical field of biological medicine engineering. According to the epitope polypeptide CD44-P2 based on the prostatic cancer stem cell marker CD44 and the application of the epitope polypeptide CD44-P2, the epitope polypeptide based on the prostatic cancer stem cell surface marker CD44 is designed and synthesized by a bioinformatics means, can sacrificially activate cytotoxic T lymphocytes, has good killing effects, canbe used for developing therapeutic peptide vaccines targeting prostatic cancer stem cells, a new technical scheme is provided for precise immunotherapy of malignant prostatic cancer, prostatic cancerstem cells can be specifically targeted, and relapse of prostatic cancer is reduced fundamentally.

Description

Technical field [0001] The present invention relates to the technical field of biomedical engineering, in particular to an epitope polypeptide CD44-P2 based on the prostate cancer stem cell marker CD44 and its application. Background technique [0002] At present, the incidence and mortality of prostate cancer are increasing year by year, and it has become the number one killer that seriously threatens human health. Although the surgical treatment, chemotherapy, radiotherapy and molecular targeted therapy of prostate cancer continue to make new progress, the effective treatment of prostate cancer is still a key issue that needs to be resolved. [0003] In recent years, as people continue to reveal the molecular and immune mechanisms of prostate cancer, prostate cancer-specific immunotherapy has received increasing attention, and obtaining an ideal tumor antigen is the key to tumor immunotherapy. With the rapid development of immunological theory and molecular biology methods, the ...

Claims

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Application Information

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IPC IPC(8): C07K14/705C12N15/12A61K39/00A61P35/00A61P13/08
CPCA61K39/00A61P13/08A61P35/00C07K14/70585
Inventor 王铸梁辉邓琼张颖张建文植凡胡七一张圣平
Owner 深圳市龙华区人民医院
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