The Efficiency And Functional Study Of NY-ESO-1-specific TCR Gene Transferred Human PBMC

ObjectiveNY-ESO-1-specific TCR gene was transduced into human PBMCs to improve itsability of specifically recognizing and killing tumor cells.And this result lay afoundation for further research of adoptive immunotherapy which is for NY-ESO-1.MethodspCDNA3.1-ESO-TCR plasmid was electroporately transferred into PBMCs separatedfrom healthy people in vitro and confirmed by RT-PCR. The phenotype analysis aftereletroporation was measured by flow cytometry method. The NY-ESO-1specificpeptide (p157-165) was added in the culture of electroporated PBMCs, the IFN-γlevel secreted by electroporated PBMCs was detected by ELISPOT assay. And wemoniter the potency of cytotoxicity of transferred PBMCs in vitro.Results1. The result of RT-PCR showed that NY-ESO-1specific TCR gene fragmentcan be detected in transferred PBMCs.2. The result of flocymetry showed that the efficiency of NY-ESO-1specificTCR expression in transferred PBMC obviously more than untransferredPBMC(P<0.05).3. Compared with untransferred PBMCs， transferred PBMCs sectrete moreIFN—γ after stimulated with peptide(P<0.05).4. The cytotoxicity of transferred PBMCs obviously stronger than untransferredPBMCs.Conclusions 1. The NY-ESO-1specific TCR expression in PBMCs was elevated byoptimized electroporation, which could improve the NY-ESO-1speicificcytotoxicity of human peripheral blood lymphocytes and provide thepossibility for cancer immunotherapy.2. We successfully established the method of infecting human PBMCs vialentivirus. It layed the foundation for the futher research of transferringTCR gene with lentivirus.