[Adenovirus containing c-FLIPs gene induces antiapoptosis activation of lymphocyte in vitro].
OBJECTIVE
To investigate the efficacy and mechanisms of recombinant adenovirus containing c-FLIPs (cellular FLICE-inhibitory protein short) in inducing antiapoptosis activation of lymphocyte in vitro.


METHODS
c-FLIPs gene was cloned from total RNA of lymphocyte with RT-PCR methods, then constructed recombined adenovirus Ad.c-FLIPs by pAdeasy system. H9 cells infected with Ad.c-FLIPs produced antiapoptosis derived from anti-Apo-1 antibody, and those antiapoptosis evaluated with PI stain flow cytometry (FCM) and Hoechst stain.


RESULTS
c-FLIPs gene was successfully cloned from lymphocytes and incorporated into recombinant adenovirus Ad.c-FLIPs.After exposure to anti-Apo-1 for 24 h, the apoptosis of H9 cells preconditioned by Ad.c-FLIPs for 24 h as measured by Hochst stain or FCM significantly decreased, compared with non-preconditioned cells, 3.60% +/- 0.21% vs. 48.33% +/- 7.41%, respectively, indicating that Ad.c-FLIPs preconditioning protected H9 cells against apoptosis induced by anti-Apo-1.


CONCLUSIONS
Recombinant adenovirus Ad.c-FLIPs effectively induced anti-apoptosis activation of lymphocyte.
Feng
Zi-yi
ZY
Department of Hepatobillary Surgery, Peking University Shenzhen Hospital, Shenzhen 518036, China.
Li
Bao-jin
BJ
Liu
Xiao-ping
XP
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000502:physiology*; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000235:genetics / Q000502:physiology*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D019014:fas Receptor / Q000032:analysis
D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; C506386:CFLAR protein, human; D019014:fas Receptor