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Antigen epitope polypeptide CD44-P3 based on prostate cancer stem cell marker CD44 and application thereof

A CD44-P3, antigen epitope technology, applied in the direction of CD44, receptor/cell surface antigen/cell surface determinant, application, etc., can solve the problem of easy tumor recurrence, low specificity of anti-tumor treatment technology, limited killing effect, 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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  • Antigen epitope polypeptide CD44-P3 based on prostate cancer stem cell marker CD44 and application thereof
  • Antigen epitope polypeptide CD44-P3 based on prostate cancer stem cell marker CD44 and application thereof
  • Antigen epitope polypeptide CD44-P3 based on prostate cancer stem cell marker CD44 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0081] 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:

[0082] MDKFWWHAAWGLCLVPLSLAQIDLNITCRFAGVFHVEKNGRYSISRTEAADLCKAFNSTLPTMAQMEKALSIGFETCRYGFIEGHVVIPRIHPNSICAANNTGVYILTSNTSQYDTYCFNASAPPEEDCTSVTDLPNAFDGPITITIVNRDGTRYVQKGEYRTNPEDIYPSNPTDDDVSSGSSSERSSTSGGYIFYTFSTVHPIPDEDSPWITDSTDRIPATTLMSTSATATETATKRQETWDWFSWLFLPSESKNHLHTTTQMAGTSSNTISAGWEPNEENEDERDRHLSFSGSGIDDDEDFISSTISTTPRAFDHTKQNQDWTQWNPSHSNPEVLLQTTTRMTDVDRNGTTAYEGNWNPEAHPPLIHHEHHEEEETPHSTSTIQATPSSTTEETATQKEQWFGNRWHEGYRQTPKEDSHSTTGTAAASAHTSHPMQGRTTPSPEDSSWTDFFNPISHPMGRGHQAGRRMDMDSSHSITLQPTANPNTGLVEDLDRTGPLSMTTQQSNSQSFSTSHEGLEEDKDHPTTSTLTSSNRNDVTGGRRDPNHSEGSTTLLEGYTSHYPHTKESRTFIPVTSAKTGSFGVTAVTVGDSNSNVNRSLSGDQDTFHPSGGSHTTHGSESDGHSHGSQEGGANTTSGPIRTPQIPEWLIILASLLALALILAVCIAVNSRRRCGQKKKLVINSGNGAVEDRKPSGLN...

Embodiment 2

[0090] The cytotoxic effect of CD44-induced effector T cells on human prostate cancer cells LNCAP-multisphere was studied by using the antigenic epitope polypeptide CD44-P3 of the CD44 protein epitope provided in Example 1 of the present invention. Specifically include 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 2 centrifuge tubes, centrifuge at 2000 r / min for 5 min, discard the supernatant, mix the precipitated cells, 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 slowly transfer the blood cell suspension to the surface of the lymphocyte separation solution with a dropper, so that a clear interface is formed between the two.

[0092] 2. After centrifuging the above-mentioned centrifuge tube with a clear inter...

Embodiment 3

[0106] Using the antigenic epitope polypeptide CD44-P3 of the CD44 protein epitope provided in Example 1 of the present invention, the cytotoxic effect of CD44-induced effector T cells on human prostate cancer cell VCaP-multicellular spheres was studied.

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

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

[0109] 3. The preparation of target cells VCaP-multicellular sphere cells and the detection method of killing effect are the same as those in Example 2.

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

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Abstract

The invention provides an antigen epitope polypeptide CD44-P3 based on a prostate cancer stem cell marker CD44 and application thereof, and relates to the technical field of biological medicine engineering. According to the antigen epitope polypeptide CD44-P3 based on the prostate cancer stem cell marker CD44 and the application thereof provided by the invention, an antigen epitope polypeptide isdesigned and synthesized by adopting bioinformatics means on basis of the prostate cancer stem cell surface-marker CD44. The synthesized antigen epitope polypeptide is capable of specifically activating cytotoxic T lymphocytes, and has good killing effect; and thus, the antigen epitope polypeptide can be used for developing therapeutic peptide vaccines targeting prostate cancer stem cells, therebyproviding new technical schemes for precise immunotherapy of malignant prostate cancer. In addition, specific targeting on prostate cancer stem cells is also enabled so as to fundamentally reduce recurrence of prostate cancer.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to an antigenic epitope polypeptide CD44-P3 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. Despite the continuous progress in surgical treatment, chemotherapy, radiotherapy and molecular targeted therapy of prostate cancer, the effective treatment of prostate cancer is still a key problem to be solved urgently. [0003] In recent years, with the continuous disclosure of the molecular mechanism and immune mechanism of prostate cancer, prostate cancer-specific immunotherapy has received increasing attention, and obtaining ideal tumor antigens is the key to tumor immunotherapy. With the rapid development of immunological theoretical methods and molecul...

Claims

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Application Information

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IPC IPC(8): C07K14/705C12N15/12C12N15/63C12N1/21C12N1/15C12N1/19C12N5/10A61K39/395A61P35/00
CPCA61K39/00A61P35/00C07K14/70585C12N15/63
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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