【摘要】 目的通过检测树突状细胞(DC)黏附HCV后的功能变化,探索CLEC4M介导HCV黏附DC的可能机制,初步揭示CLEC4M对DC介导的固有免疫应答的影响。方法提取正常人外周血单个核细胞,并诱导分化为成熟DC,流式细胞术检测DC表面CLEC4M的表达情况。利用制备好的HCVcc病毒液感染成熟DC,并设置阳性对照组、甘露聚糖干扰组、CLEC4M单抗干扰组和空白对照组,分别收集感染各组的细胞上清液,采用ELISA法检测各组细胞上清液中IL-10与IL-12p70的水平变化。计量资料多组间比较采用单因素方差分析,进一步两组间比较采用LSD-t检验。结果 DC表面CLEC4M的表达阳性率高达82.6%。ELISA法检测各组细胞上清IL-10、IL-12p70的水平,4组间比较差异有统计学意义(F值分别为54.46、77.72,P值均<0.001),其中阳性对照组中IL-10、IL-12p70的表达水平明显高于两干扰组(P值均<0.001),阳性对照组、两干扰组IL-10、IL-12p70的水平均高于空白对照组(P值均<0.001)。结论 CLEC4M介导了HCV黏附DC,参与机体的免疫应答并调节DC免疫分子的表达。更多还原

【Abstract】 Objective To investigate the changes in the function of dendritic cells(DCs) after HCV adherence,possible mechanisms of HCV adherence of DCs mediated by CLEC4 M,and the impact of CLEC4 M on innate immune response mediated by DCs. Methods Peripheral blood mononuclear cells(PBMCs) were isolated from healthy people and induced to differentiate into mature DCs. Flow cytometry was used to measure the expression of CLEC4 M on the surface of DCs. Mature DCs were infected by HCVcc and positive control group,mannan interference group,CLEC4 M monoclonal antibody interference group,and blank control group were established. The supernatant was collected from each infected group,and ELISA was used to measure the changes in the levels of interleukin-10(IL-10) and interleukin-12p70(IL-12p70) in supernatant. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between any two groups. Results The positive rate of CLEC4 M in the surface of DCs reached 82. 6%. There were significant differences in the levels of IL-10 and IL-12p70 between the groups(F =54. 46 and 77. 72,both P < 0. 001). The positive control group had significantly higher levels of IL-10 and IL-12p70 than the two interference groups(all P < 0. 001),and compared with the blank control group,the positive control group,CLEC4 M monoclonal antibody interference group,and mannan interference group had significantly higher levels of IL-10 and IL-12p70(all P < 0. 001). Conclusion CLEC4 M mediates HCV adherence of DCs,participates in the body’s immune response,and regulate the expression of DC immune molecules.更多还原