[Construction and biological activities of CTLA4Ig adenovirus vectors].
OBJECTIVE
To construct CTLA4Ig adenovirus vectors (AdCTLA4Ig) by homologous recombination and study their activity, and to employ the vectors to induce cardiac transplantation tolerance by gene therapy.


METHODS
CTLA4Ig gene was cloned to pCA13 adenovirus shuttle plasmid by recombination strategy. Construction of CTLA4Ig adenovirus vectors was performed by homologous recombination of pCA13 plasmid containing CTLA4Ig gene with adenovirus helper plasmid, followed by packaged with 293 cells. Expression and secretion of CTLA4Ig was confirmed by RT-PCR, SDS-PAGE and Western blot. The inhibitory effect of supernate of 293 cells infected with AdCTLA4Ig on MLR in-vitro was observed. A biological activity of CTLA4Ig adenovirus vectors was determined by AdCTLA4Ig gene therapy in rats in-vivo.


RESULTS
The Construction of CTLA4Ig adenovirus vector was successful. It was confirmed that the supernatant of 293 cells infected with AdCTLA4Ig could inhibit MLR in-vitro. It was also showed that CTLA4Ig adenovirus vectors could induce transplantation tolerance and prolong allograft survival when they were administrated in rats in-vivo.


CONCLUSIONS
CTLA4Ig adenovirus vectors successfully constructed can infect 293 package cells and secrete CTLA4Ig. The CTLA4Ig protein can inhibit T cell activation. The CTLA4Ig adenovirus vectors can be employed to gene therapy in-vivo, and induce transplantation tolerance.
Zhang
Qing-yin
QY
Department of Immunology, Peking University Health Science Center, Beijing 100083, China.
Jin
Yong-zhu
YZ
Zhang
Hai-bin
HB
Xie
Shu-sheng
SS
D000069594:Abatacept; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004622:Embryo, Mammalian; D015316:Genetic Therapy; D005822:Genetic Vectors; D016027:Heart Transplantation; D006801:Humans; D018796:Immunoconjugates / Q000235:genetics / Q000378:metabolism*; D007668:Kidney / Q000166:cytology*; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D009206:Myocardium / Q000378:metabolism; D010957:Plasmids; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D023001:Transplantation Tolerance
D018796:Immunoconjugates; D011993:Recombinant Fusion Proteins; D000069594:Abatacept