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Antigen epitope polypeptide cd44-p2 based on prostate cancer stem cell marker cd44 and its application

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

Inactive Publication Date: 2020-09-15
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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  • Antigen epitope polypeptide cd44-p2 based on prostate cancer stem cell marker cd44 and its application
  • Antigen epitope polypeptide cd44-p2 based on prostate cancer stem cell marker cd44 and its application
  • Antigen epitope polypeptide cd44-p2 based on prostate cancer stem cell marker cd44 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Design and synthesis of embodiment 1 CD44 epitope short peptide

[0077] 1. Find the complete amino acid sequence (NP_000601.3) of human CD44 protein from the international open shared gene bank NCBI Genbank, a total of 742 amino acids. Its amino acid sequence is as follows (SEQ ID NO: 3):

[0078] MDKFWWHAAWGLCLVPLSLAQIDLNITCRFAGVFHVEKNGRYSISRTEAADLCKAFNSTLPTMAQMEKALSIGFETCRYGFIEGHVVIPRIHPNSICAANNTGVYILTSNTSQYDTYCFNASAPPEEDCTSVTDLPNAFDGPITITIVNRDGTRYVQKGEYRTNPEDIYPSNPTDDDVSSGSSSERSSTSGGYIFYTFSTVHPIPDEDSPWITDSTDRIPATTLMSTSATATETATKRQETWDWFSWLFLPSESKNHLHTTTQMAGTSSNTISAGWEPNEENEDERDRHLSFSGSGIDDDEDFISSTISTTPRAFDHTKQNQDWTQWNPSHSNPEVLLQTTTRMTDVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTSTIQATPSSTTEETATQKEQWFGNRWHEGYRQTPKEDSHSTTGTAAASAHTSHPMQGRTTPSPEDSSWTDFFNPISHPMGRGHQAGRRMDMDSSHSITLQPTANPNTGLVEDLDRTGPLSMTTQQSNSQSFSTSHEGLEEDKDHPTTSTLTSSNRNDVTGGRRDPNHSEGSTTLLEGYTSHYPHTKESRTFIPVTSAKTGSFGVTAVTVGDSNSNVNRSLSGDQDTFHPSGGSHTTHGSESDGHSHGSQEGGANTTSGPIRTPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVIN...

Embodiment 2

[0087] The antigenic epitope polypeptide CD44-P2 of the CD44 protein antigenic 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 LNCAP-multicellular spheres. Specifically include the following steps:

[0088] 1. Separation and preparation of peripheral blood mononuclear cells: collect 10 mL of peripheral blood collected from HLA-A2 positive healthy volunteers into 2 centrifuge tubes, centrifuge at 2000 r / min for 5 min, discard the supernatant, mix the precipitated cells, add physiological Add 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 medium, and slowly transfer the blood cell suspension to the surface of the lymphocyte separation medium with a dropper, so that a clear interface is formed between the two.

[0089] 2. After centrifuging the above-mentioned centrifuge ...

Embodiment 3

[0102] The antigenic epitope polypeptide CD44-P2 of the CD44 protein antigenic 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.

[0103] 1. The separation and preparation method of peripheral blood mononuclear cells is the same as in Example 2.

[0104] 2. The culture method of DC-CIK cells is the same as that in Example 2.

[0105] 3. The preparation of target cells VCaP-multicellular spheroid cells and the detection method of killing effect are the same as in Example 2.

[0106] Such as Figure 5A and Figure 5B As shown, the CIK cells induced by the CD44 epitope peptide have a significant killing effect on the VCaP-multicellular spheres enriched with human prostate cancer cell stem cells. When the effect-to-target ratio is 5:1, the average killing effect is 8.9% of the background. % increased to 23.1%; when the effect-to-target ratio was 1...

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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 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 morbidity 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 have made new progress, the effective treatment of prostate cancer is still a key problem to be solved. [0003] In recent years, with the continuous revelation of the molecular mechanism and immune mechanism of prostate cancer, more and more attention has been paid to the specific immunotherapy of prostate cancer, and the key to tumor immunotherapy is to obtain an ideal tumor antigen. With the rapid development of immunological theoretical methods and molecular...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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