Fallopian tube cancer target antigen, CTL cell cultured by stimulating fallopian tube cancer target antigen and application of fallopian tube cancer target antigen
A fallopian tube cancer, stimulating culture technology, applied in the field of tumor treatment, can solve the problems such as the inability to meet the individuality and variability of tumors
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[0032] The present invention also provides the preparation method of said CTL cells, comprising the following steps: 1) adjusting the concentration of CTL cells to 2-3×10 5 cells / mL; 2) Mix the CTL cells after adjusting the cell concentration, IL-2 and the fallopian tube cancer target antigen combination to obtain a culture system; 3) culture the culture system to obtain the fallopian tube cancer target antigen combination stimulation culture CTL cells.
[0033] In the present invention, the CTL cells are preferably obtained by co-culturing PBMC cells and APC cells. In the present invention, the step of culturing the APC cells is preferably as follows: PBMC cells are resuspended and cultured with fallopian tube cancer target antigen solution to obtain them. In the present invention, the culture medium of the recovery culture is preferably 1640+10% (v / v) FBS; the time of the recovery culture is preferably 22~26h, more preferably 24h; the temperature of the recovery culture is ...
Embodiment 1
[0043] For the whole exome sequencing results of tumor tissue, the tumor neoantigen analysis was performed using ImmunoMining tumor individualized neoantigen analysis and prediction software. The software copyright registration number: 2019SR0130720. Dingcheng Peptide Source Bio-Information Technology Co., Ltd. obtained 50 predicted antigens; synthesized the predicted 50 antigens (synthesized by the Polypeptide Synthesis Department of Nanjing GenScript).
[0044] Table 1 Peptide sequence and corresponding number
[0045]
[0046]
[0047]
[0048] 1. Preparation of APC by co-culture:
[0049] 1) Resuscitated frozen PBMC (2.5×10 7 cells), dilute the freezing solution with 1640+10% FBS, centrifuge at 1500rpm for 5min, resuspend with 10mL 1640+10%FBS, 1×10 6 cells / mL, 37°C, 5% CO 2 Recovery culture for 24h;
[0050] 2) Antigen polypeptide solutions were prepared separately, 1640+10% FBS+200U / mL IL-2+1600U / mL GM-CSF, the final concentration of each antigen polypeptide...
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