Adenovirus containing c-FLIPs gene induces antiapoptosis activation of lymphocyte in vitro.
- Author:
Zi-yi FENG
1
;
Bao-jin LI
;
Xiao-ping LIU
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Apoptosis; physiology; CASP8 and FADD-Like Apoptosis Regulating Protein; genetics; physiology; CD4-Positive T-Lymphocytes; cytology; metabolism; Cell Line; Flow Cytometry; Humans; Lymphocytes; cytology; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Transfection; fas Receptor; analysis
- From: Chinese Journal of Experimental and Clinical Virology 2005;19(2):114-117
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the efficacy and mechanisms of recombinant adenovirus containing c-FLIPs (cellular FLICE-inhibitory protein short) in inducing antiapoptosis activation of lymphocyte in vitro.
METHODSc-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.
RESULTSc-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.
CONCLUSIONRecombinant adenovirus Ad.c-FLIPs effectively induced anti-apoptosis activation of lymphocyte.
