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Antigen epitope peptide CD44-P1 based on prostatic cancer stem cell marker CD44 and application thereof

A technology of CD44-P1 and epitope, which is applied in the direction of CD44, receptor/cell surface antigen/cell surface determinant, cancer antigen components, etc., can solve the problem of limited killing effect, easy tumor recurrence, and specificity of anti-tumor treatment technology Low-level problems, to achieve the effect of reducing recurrence, enhancing the inhibitory and clearing effect, and good killing effect

Inactive Publication Date: 2019-04-09
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 peptide CD44-P1 based on prostatic cancer stem cell marker CD44 and application thereof
  • Antigen epitope peptide CD44-P1 based on prostatic cancer stem cell marker CD44 and application thereof
  • Antigen epitope peptide CD44-P1 based on prostatic cancer stem cell marker CD44 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Design and synthesis of embodiment 1 CD44 epitope short peptide

[0083] 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:

[0084] MDKFWWHAAWGLCLVPLSLAQIDLNITCRFAGVFHVEKNGRYSISRTEAADLCKAFNSTLPTMAQMEKALSIGFETCRYGFIEGHVVIPRIHPNSICAANNTGVYILTSNTSQYDTYCFNASAPPEEDCTSVTDLPNAFDGPITITIVNRDGTRYVQKGEYRTNPEDIYPSNPTDDDVSSGSSSERSSTSGGYIFYTFSTVHPIPDEDSPWITDSTDRIPATTLMSTSATATETATKRQETWDWFSWLFLPSESKNHLHTTTQMAGTSSNTISAGWEPNEENEDERDRHLSFSGSGIDDDEDFISSTISTTPRAFDHTKQNQDWTQWNPSHSNPEVLLQTTTRMTDVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTSTIQATPSSTTEETATQKEQWFGNRWHEGYRQTPKEDSHSTTGTAAASAHTSHPMQGRTTPSPEDSSWTDFFNPISHPMGRGHQAGRRMDMDSSHSITLQPTANPNTGLVEDLDRTGPLSMTTQQSNSQSFSTSHEGLEEDKDHPTTSTLTSSNRNDVTGGRRDPNHSEGSTTLLEGYTSHYPHTKESRTFIPVTSAKTGSFGVTAVTVGDSNSNVNRSLSGDQDTFHPSGGSHTTHGSESDGHSHGSQEGGANTTSGPIRTPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVINSGNGAVEDRKPSGLN...

Embodiment 2

[0092] The antigenic epitope polypeptide CD44-P1 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:

[0093] 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.

[0094] 2. After centrifuging the above-mentioned centrifug...

Embodiment 3

[0108] The antigenic epitope polypeptide CD44-P1 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.

[0109] 1. The separation and preparation method of peripheral blood mononuclear cells is the same as in Example 2.

[0110] 2. The culture method of DC-CIK cells is the same as that in Example 2.

[0111] 3. The preparation of target cells VCaP-multicellular spheroid cells and the detection method of killing effect are the same as in Example 2.

[0112] Such as Figure 6A and Figure 6B As shown, by comparative analysis of flow cytometry, the results showed that the proportion of CIK cells activated by the CD44 epitope polypeptide was significantly increased; at day 0 ( Figure 6A ), the results of flow cytometry analysis showed that the proportion of CD3 positive T cells was 65.20%, the proportion of CD4 positive ...

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Abstract

The invention provides antigen epitope peptide CD44-P1 based on a prostatic cancer stem cell marker CD44 and application thereof, and relates to the technical field of biomedicine engineering. The antigen epitope peptide based on the prostatic cancer stem cell marker CD44 is designed and synthesized through bioinformatics means, specifically activates cytotoxicity T lymphocyte, has an excellent kill effect, can be applied to the development of prostatic cancer stem cell targeted therapeutic peptide vaccine, provides a novel technical scheme for the precise immunological therapy of malignant prostatic cancer, and can further specifically target prostatic cancer stem cell to reduce the recurrence of prostatic cancer.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to an epitope polypeptide CD44-P1 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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IPC IPC(8): C07K14/705C12N15/12A61K39/00A61P35/00
CPCA61K39/0011A61P35/00C07K14/70585
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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