[Construction and identification of recombinant adenovirus vector harboring CTLA4Ig-IRES 2-Ikappa-Balpha gene in ECV-304 cells].
OBJECTIVE
To construct adenovirus vector harboring CTLA4Ig-IRES 2-IkappaBalpha gene,and investigate its expression in ECV304 cells.


METHODS
Recombinant adenovirus vector harboring CTLA4Ig-IRES 2-IkappaBalpha was constructed by homologous recombination in E.coli BJ5183. Then recombinant vector was packaged and propagated in 293 cells. ECV304 cells were infected with the recombinant adenovirus, and Western Blot was used to detect IkappaBalpha and CTLA4Ig protein expression in infected ECV304 cells. The effect of expressed IkappaBalpha and CTLA4Ig on the expression of TNF-alpha of ECV304 cells stimulated with LPS was also investigated.


RESULTS
The recombinant CTLA4Ig-IRES 2-IkappaBalpha adenovirus was generated by homologous recombination and identified by PCR methods. Ikappa-Balpha and CTLA4Ig were expressed in ECV304 cells infected with the recombinant adenovirus. The production of TNF-alpha induced by the LPS was inhibited in infected ECV304 cells.


CONCLUSIONS
The recombinant CTLA4Ig-IRES 2-IkappaBalpha adenovirus may offer a method to down regulate the inflammatory cytokines in the inflammatory reaction which may lay the foundation for genetherapy of immunosuppression.
Li
Ning
N
Department of Immunology, Medical School of Xi'an Jiaotong University, Xi'an 710061, China.
Yuan
Yu-Kang
YK
Wu
Jun
J
D000069594:Abatacept; D000256:Adenoviridae / Q000235:genetics*; D002460:Cell Line; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics*; D018796:Immunoconjugates / Q000235:genetics*; D007165:Immunosuppression Therapy; D000072000:NF-KappaB Inhibitor alpha
D004274:DNA, Recombinant; D020672:I-kappa B Proteins; D018796:Immunoconjugates; C000606204:NFKBIA protein, human; D000072000:NF-KappaB Inhibitor alpha; D000069594:Abatacept; D004262:DNA Restriction Enzymes