【摘要】 目的探讨体外丙型肝炎病毒(HCV)感染对NK细胞功能产生的影响及其可能的机制。方法利用质粒JC1-Flag2体外转录得到的HCV感染性颗粒(HCVcc)以MOI4.8感染Huh7.5细胞(Huh7.5-HCVcc),与健康人外周血分离得到的NK细胞进行共培养。与Huh7.5-HCVcc细胞共培养前后,采用ELISA法和MTT比色法分别检测NK分泌细胞因子IFN-γ、TNF-α和IL-10的水平和细胞杀伤活性,以评估HCV感染对NK细胞功能的影响。结果与MOI4.8的Huh7.5-HCVcc细胞共培养,NK细胞分泌IFN-γ、TNF-α和IL-10水平受到抑制,其中IFN-γ在共培养6 h(P<0.001)、9 h(P<0.001)、12 h(P<0.001)受到显著抑制,TNF-α在共培养6 h(P<0.001)、9 h(P<0.001)、12 h(P=0.001)受到显著抑制,IL-10在共培养6 h(P<0.001)、9 h(P=0.006)受到显著抑制;且NK细胞分泌细胞因子IFN-γ、TNF-α和IL-10的功能均在共培养6 h受到的抑制效应最大,平均抑制率分别为24.1%、20.7%和24.3%。NK细胞杀伤活性在共培养6 h(P=0.023)受到抑制,平均抑制率为16.6%。结论在体外与MOI4.8的Huh7.5-HCVcc细胞共培养,NK细胞分泌细胞因子(IFN-γ、TNF-α和IL-10)和细胞杀伤功能均受到抑制,且在不同时间点受到的抑制程度不同,提示NK细胞在HCV感染的不同阶段可能起到不同的作用。更多还原

【Abstract】 Objective The numbers,subsets and function of natural killer( NK) cells altered in chronic hepatitis C virus( HCV)infected patients which indicating its association with chronic HCV infection,but the exact mechanisms remain unclear. To explore the functional effect of HCV infection on NK cells in vitro and the possible mechanism behind it. Methods Human hepatoma Huh7.5 cells were transfected with genomic RNA from cell culture adapted JC1 genotype 2a HCV strain( Huh7. 5-HCVcc) and cocultured with NK cells isolated from whole peripheral blood of concentrated healthy donors. The functional effect of HCV infection on NK cells was assessed by testing cytokine-secreting and cytotoxicity before and after co-culture. Results When co-cultrued with Huh7. 5-HCV of MOI 4. 8,the levels of IFN-γ,TNF-α and IL-10 secreted by NK cells were inhibited. The inhibition of secreting IFN-γ was specially significant at 6( P < 0. 001),9( P < 0. 001) and 12( P = 0. 001) hours co-cultured with Huh7. 5-HCV. The inhibition of secreting TNF-α was specially significant 6 h( P < 0. 001),9 h( P < 0. 001) and 12 h( P = 0. 001) after co-cultured with Huh7. 5-HCVcc. And the significant inhibition of secreting IL-10 appeared at 6 h( P < 0. 001) and 9 h( P = 0. 006) after cocultured with Huh7. 5-HCVcc. The strongest cytokine-secreting inhibition appeared 6 h after co-cultured with Huh7. 5-HCVcc,the inhibition rate was 24. 1%,20. 7% and 24. 3%,respectively. The cytotoxicity decreased significantly 6 h after co-cultured with Huh7. 5-HCVcc( P = 0. 023),and the average inhibition rate was 16. 6%. Conclusion When in vitro cocultrued with Huh7. 5-HCVcc of MOI 4. 8,the NK cells functions of IFN-γ,TNF-α and IL-10 cytokine-secreting and cytotoxicity were inhibited. The degree of inhibition was different at different time points,which indicated that NK cells may act differently during HCV infection.更多还原