Objective    To construct a eukaryotic expression vector co-expressing genes encoding

T-bet

and

c-FLIP

and to detect its bioactivity in transfected PBMCs.     Methods   

c-FLIP

and

T-bet

genes were cloned from tota

l RNA of lymphocyte with RT-PCR methods

then constructed into the eukaryotic expression vector by using pIRES and pEGFP-C1 vectors. Lymphocyte from blood donated by hepatocellular carcinoma patients infected by pEGFPT-bet-IRES-c-FLIP with QuikFusin Transfection Kit induced the Th1/Th2 shift and produced anti-apoptosis derived from CH-11 antibody

whose anti-apoptosis evaluated with PI stain flow cytometry (FCM).     Results   

T-bet

and

c-FLIP

genes were successfully cloned from lymphocytes and incorporated into the eukaryotic expression vector. Exposed to CH-11 for 24 h

the apoptosis of lymphocyte preconditioned by pEGFPT-bet-IRES-c-FLIP for 24 h was significantly decreased

compared with no-preconditioned cells (5.73%±0.37%

vs

50.12%±8.02%)

indicating that pEGFPT-bet-IRES-c-FLIP preconditioning protected lymphocyte against apoptosis induced by CH-11. Lymphocyte infected by pEGFPT-bet-IRES-c-FLIP significantly enhanced IFN-γ levels and decreased IL-4 levels

compared with no-preconditioned cells and the cells infected by empty vector.     Conclusion    A eukaryotic expression vector co-expressing genes encoding

T-bet

and

c-FLIP

may induce anti-apoptotic activities of lymphocyte and Th1 differentiation.