Effects on macrophages activity transfected transiently with pcDNA3-HBV / 中华传染病杂志

ABSTRACT

Objective To explore the mechanism of immune tolerance in chronic B hepatitis, the effects of HBV infection on the functions of macrophages and related mechanism of signal transduction by transfection in vitro were studied. Methods Peritoneal macrophages of mice were isolated regularly and cultured, transfected transiently with pcDNA3-HBV or pcDNA3 plasmid DNA and cultured under the stimulation of LPS. After 72 h, RT-PCR was performed to detect the expression of PreS1, TNF-? or IL-1? mRNA. To detect the expression of NF-?B RelA protein by FCM, and the level of nitric oxide in cultural supernatant was measured with Griess reaction. Results After being transfected with pcDNA3-HBV,peritoneal macrophages had the expression of PreS1 mRNA, but have lower level of TNF-?、IL-1? mRNA and transcriptional factor NF-?B, compared with pcDNA3-transfected control group; the level of nitric oxide in pcDNA3-HBV group was also decreased. Conclusions Transient transfection of pcDNA3-HBV could decrease the function of macrophages directly by inhibiting NF-?B activity and effector molecules production, which may be one of the mechanisms of immune tolerance in chronic B hepatitis.