HCV感染人滋养层细胞过程中抗体依赖性增强作用的初步研究  

The study on the antibody-dependent enhancement of HCV infection in the human cytotrophoblasts cultured in vitro

作　　者：聂青和[1] 罗新栋[1] 张亚飞[1] 黄晓峰[2] 李军[1] 邵彬[1] 程勇前[1] 陈朝霞[1] 

机构地区：[1]第四军医大学唐都医院传染病科,全军感染病诊疗中心,西安710038 [2]第四军医大学电子显微镜室

出　　处：《胃肠病学和肝病学杂志》2006年第4期379-382,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(No:39970767)

摘　　要：目的研究丙型肝炎病毒(HCV)在感染人滋养层细胞的过程中是否存在抗体依赖性感染增强(ADE)作用,探讨HCV母婴传播的分子机制。方法将HCV阳性血清以4种不同方式感染体外培养人滋养层细胞,应用RT-PCR法对标本进行HCVRNA定性检测,并进一步评估不同浓度CD16单抗对感染过程的阻断作用。此外,应用免疫电镜技术观察滋养层细胞感染HCV病毒颗粒情。结果HCVRNA在全血清组、CD81单抗阻断组及LDL-R单抗阻断组多个标本均为阳性表达,且平均拷贝数较高,在FcγRⅢ(CD16)单抗阻断组多个标本内未能检测到HCVRNA的表达,且阳性标本拷贝数较低;CD16单抗对感染的阻断作用随浓度的升高而增强;在全血清组、CD81单抗阻断组及LDL-R单抗阻断组多个标本细胞内可间断测得未脱壳的HCV病毒颗粒,CD16单抗阻断组细胞内未能检出。结论滋养层细胞膜CD81和LDL-R分子不参与HCV感染滋养层细胞的过程,而FcγRⅢ分子则与此过程密切相关。HCV感染人滋养层细胞过程中存在抗体依赖性增强作用。Objective To assess the antibody-dependent enhancement （ADE） of HCV infection in the human cytotrophoblasts cultured in vitro. Methods Cytotrophoblasts cultured in vitro were incubated with DMEM-HG medium with HCVRNA-positive serum in four different ways. Quantity RT-PCR was used to assess the expression of HCV RNA in cytotrophoblasts. And the blocking assay of FcR-ADE was processed by pretreating the cytotrophoblasts with different titer of antiFcγR Ⅲ （CD16） McAb. Immunoelectron microscopy was used to observe the HCV particles in HCV infected cytotrophoblasts. Results HCV RNA was detected in most samples of serum control group, anti-CD81 McAb pretreated group and anti-LDL-R McAb pretreated group, and the average copies of each of the three groups were high; In anti-CD16 McAb pretreated group, few samples were detected HCV RNA and even in the positive results, the copies were low. Furthermore, increasing anti- CD16 McAb titer used in the assay of FcR-ADE enhanced the block effection. HCV-like particles conjoined with golden granules could be obseverd in serum control group, anti-CD81 McAb pretreated group and anti-LDL-R McAb pretreated group, while not in anti-CD16 McAb pretreated group. Conclusion CD81 and LDL-R don＇t participate in the process of HCV infecting cytotrophoblasts, while FcγR Ⅲ is correlative with the process. ADE may contribute to the HCV infection in cytotro- phoblasts.

关 键 词：丙型肝炎病毒 抗体依赖性感染增强作用 母婴传播 人滋养层细胞 体外感染 

分 类 号：R512.6[医药卫生—内科学]