人正常肝细胞LO2感染HBV后表达CCL22

Expression of CCL22 in human normal liver LO2 cells after infected with hepatitis B virus

目的探讨感染HBV的人正常肝细胞LO2表达趋化因子CCL22的机制。方法首先用HBV病毒感染LO2细胞,通过ELISA检测细胞IL-32表达情况;之后用外源人类重组IL-32蛋白刺激人急性单核细胞白血病细胞THP-1或用HBV感染共培养的LO2和THP-1细胞,通过ELISA检测TNF-α表达情况;其次用外源人类重组TNF-α蛋白刺激LO2细胞或用HBV感染共培养的LO2和THP-1细胞,使用特异性抗体通过Western blot检测CREB蛋白磷酸化情况,并通过ELISA检测趋化因子CCL22的表达情况。结果 HBV诱导LO2细胞表达IL-32;病毒诱导的IL-32诱导THP-1细胞分泌TNF-α(P<0.05);TNF-α诱导LO2细胞活化CREB通路(P<0.05);CREB通路活化后促使LO2表达趋化因子CCL22(P<0.05)。结论 HBV感染与THP-1共培养的LO2细胞可表达CCL22,其机制可能是HBV诱导LO2细胞表达IL-32,之后IL-32诱导THP-1细胞分泌TNF-α,进而TNF-α促进HBV感染的LO2细胞表达趋化因子CCL22。

Objective To investigate the mechanism of expression of the chemokine CCL22 in human normal liver LO2 cells after infected with hepatitis B virus(HBV).Methods LO2 cells were first infected with HBV virus and the expression of IL-32 was detected by ELISA.Then,exogenous human recombinant IL-32 protein was used to stimulate human acute leukemia cell line THP-1 or HEB and THP-1 cells co-cultured with HBV,and TNF-α expression was detected by ELISA.Second,exogenous human recombinant TNF-α protein was applied for stimulation or co-cultured LO2 and THP-1 cells were infected with HBV.LO2 cells were used to detect the phosphorylation of CREB protein by western blotting with specific antibody and to detect the expression of the chemokine CCL22 by ELISA.Results HBV induced IL-32 expression in LO2 cells (P< 0.05),and virus-induced IL-32 in turn induced the secretion of TNF-α by THP-1 cells (P< 0.05).TNF-α induced activation of the CREB pathway in LO2 cells.This activation then promoted the expression of the chemokine CCL22 in LO2 cells (P < 0.05).Conclusions LO2 cells co-cultured under conditions of HBV infection and in the presence of THP-1 can express CCL22.The mechanism behind this may involve HBV inducing IL-2 expression in LO2 cells,and then IL-32 induces the secretion of TNF-α by THP-1 cells,after which TNF-α promotes the HBV-infection.LO2 cells to express chemokine CCL22.