论文部分内容阅读
Objective To investigate whether TNF-α gene-modified Tca8113 cells (Tca8113/TNF-α) canbe used as vaccine for oral squamous cell carcinoma. Methods TNF-α gene was transduced into Tca8113 cellsin vitro with retroviral vector carring genes for both TNF-α and NeoR. After that, presence and expression of exoge-nous gene in the transgenic cells, expression of HLA antigen on the cells, expression of TNF-α and survival rate ofthe cells after irradiation and cryopreservation, and mutagenic activity of the cells were analyzed by PCR technique,EL1SA technique, FACS technique, 60Co irradiation inactivation test, cryopreservation test, and Ames test, respec-tively. Results The presence of both TNF-a and NeoR gene and expression of TNF-α gene were demonstrated intransgenic cells. The levels of the HLA-A, B, C, DR expressed by Tca8113/TNF-α were higher than by the parentalcells. Tca8113/TNF-α continued to secrete TNF-α for 14 d, there was a secretion peak time from d4 to d6;and, allthe cells died by dl4 after irradiation. The Level of TNF-α secreted by Tca8113/TNF-α cryopreserved for 48 h wasno different from that cryopreserved for 1 week after irradiation, the level of TNF-α secreted by the cryopreservedcells was just a little lower than that secreted by the noncryopreserved cells. Both DNA and supernatant of the cellshave no mutagenic activity. Conclusion TNF-α gene can be transduced into Tca8113 cells with retroviral vec-tor, and the cells can express TNF-α. Expression of HLA 1,11 antigens on Tca8113 cells can be increased by TNF-αgene transduction. Irradiation is a reliable inactivation method, and cryopreservation is a feasible conservationmethod for Tca8113/TNF-α. Ames test result indicate that Tca8113/TNF-α has no mutagenic activity.
Methods TNF-α gene was modified to Tca8113 cells (Tca8113 / TNF-α) canbe used as vaccine for oral squamous cell carcinoma. Methods TNF-α gene was transduced into Tca8113 cells in vitro with retroviral vector carring genes for both TNF- α and NeoR. After that, presence and expression of exoge- nous gene in the transgenic cells, expression of HLA antigen on the cells, expression of TNF- [alpha] and survival rate of the cells after irradiation and cryopreservation, and mutagenic activity of the cells by PCR technique, EL1SA technique, FACS technique, 60Co irradiation inactivation test, cryopreservation test, and Ames test, respectrively. Results The presence of both TNF-a and NeoR gene and expression of TNF-α gene were demonstrated in intragenic cells. levels of the HLA-A, B, C, DR expressed by Tca8113 / TNF-α were higher than by the parental cells. Tca8113 / TNF- α continued to secrete TNF- α for 14 d, there was a secretion peak time from d4 to d6; and, allthe cells died by dl4 after irradiation. The Level of TNF-α secreted by Tca8113 / TNF-α cryopreserved for 48 h wasno different from that cryopreserved for 1 week after irradiation, the level of TNF-α secreted by the cryopreserved cells was just a little lower than That secreted by the non cryopreserved cells. Both DNA and supernatant of the cells have no mutagenic activity. Conclusion TNF-α gene can be transduced into Tca8113 cells with retroviral vec-tor, and the cells can express TNF- Antigens on Tca8113 cells can be increased by TNF-α gene transduction. Irradiation is a reliable inactivation method, and cryopreservation is a feasible conservation method for Tca8113 / TNF-α. Ames test result indicate that Tca8113 / TNF-α has no mutagenic activity.