Effects Of Lymphocyte Co-Culture On Proliferation And Apoptosis Of Hepatitis B Virus (Hbv)-Expressing Normal Human Mesangial Cells (Nhmcs)

Objective: This study is to investigate the effects of the lymphocyte co-culture on the proliferation and apoptosis of the hepatitis B virus (HBV)-expressing normal human mesangial cells (NHMCs). Methods: NHMCs were transfected with the PHY106-CHBV plasmid containing C genotypeHBV with liposome. These cells were co-cultured with human peripheral blood lymphocytes. The HBsAg and HBeAg contents in the supernatant were detected at 24 h, 48 h, and 72 h after transfection. Cell proliferation was assessed with the MTT assay, and apoptosis was detected with flow cytometry. Results: In the normal control and negative control groups, negative results for the HBsAg and HBeAg detection were observed, at all the time points. However, in the transfected NHMCs, positive results were observed for the HBsAg and HBeAg detection at 48 h and 72 h after transfection. These HBV-expressing NHMCs were co-cultured with human peripheral blood lymphocytes for 48 h. Results from the MTT assay showed that, the proliferation of NHMCs was not significantly affected by the transfection of the empty plasmid. However, compared with the control group, the proliferation rate was dramatically declined in HBV-expressing NHMCs. Moreover, the lymphocyte co-culture further significantly decreased the proliferation of HBV-expressing NHMCs. In addition, our results from the flow cytometry showed that, the early apoptosis rate in the co-culture group was significantly higher than the HBV-expressing group. Conclusion: Co-culture of peripheral blood lymphocytes could inhibit proliferation and enhance apoptosis of HBV-expressing NHMCs. These findings might contribute to the understanding of the pathogenic mechanisms of the HBV-associated glomerulonephritis.