Abstract: Objective: NY-ESO-1-specific TCR gene was transduced into human peripheral blood lymphocytes (PBLs) to improve its ability of specifically recognizing and killing tumor cells. Method: pCDNA3.1-ESO-TCR plasmid was electroporately transferred into PBLs separated from healthy people in vitro and confirmed by RT-PCR. The phenotype analysis after eletroporation was measured by flow cytometry method. The NY-ESO-1 specific peptide (p157-165) was added in the culture of electroporated PBLs, the IFN-γ level secreted by electroporated PBLs was detected by ELISPOT assay. Result: The NY-ESO-1-specific TCR fragments in electroporated PBLs were detected by RT-PCR. The specific expression of NY-ESO-TCR in transferred PBLs is significantly higher than that in untransferred PBLs (P<0.05). The positive dots of IFN-γ secretion in transferred PBLs stimulated by peptide p157-165 was significantly more than that in untransferred PBLs (P<0.05). Conclusion: The NY-ESO-1 specific TCR expression in PBLs was elevated by optimized electroporation, which could improve the efficiency of IFN-γ secretion in human peripheral blood lymphocytes and provide the possibility for cancer immunotherapy.

Key words: TCR, PBLs, Electroporation, NY-ESO-1