HIV感染急性期NK细胞抗HBV功能增强的机制研究

慢乙肝治疗获得表面抗原及e抗原转阴非常困难，其控制及清除机制尚不清楚。基于已建立的男同性恋HIV感染队列，我们发现慢性HBV合并急性HIV感染后，机体对HBV控制和清除能力增加，获得较高表面抗原及e抗原转阴率，进一步发现NK功能增强在其中起重要作用。我们设想HIV感染急性期NK活化通路某些蛋白过表达或抑制通路某些蛋白低表达在其中发挥了重要作用。我们拟采用慢性HBV合并HIV感染前及急性期标本。首先，纯化NK，对NK功能通路相关基因利用RNA-seq技术及实时定量PCR技术进行检测及对比分析；其次，通过siRNA技术对上述差异基因进行功能验证；再次，将靶基因转染进入正常及慢乙肝病人NK细胞，与HBV感染HepaRG细胞共培养，检测NK功能及对HBV清除能力；最后，通过免疫组化检测肝中NK基因表达。进一步揭示HIV感染急性期NK抗HBV的机制，为针对NK细胞设计抗慢性HBV靶点提出更多思路。

It is very difficult to achieve the HBsAg and HBeAg seroconvertion in chronic hepatitis patients, and the mechanisms are not clear yet. Based on the men-having-sex-with-men HIV infection cohort, we found that the chronic hepatitis patients could acquire higher HBsAg and HBeAg seroconvertion during acute HIV infection. Furthermore, we discovered that the increased NK cell functions played an important role in the process. We supposed that the overexpression of proteins from NK cell activation pathway or low expression of protein from NK cell inhibition pathway in the process were extremely vital. The samples before HIV infection and acute HIV infection from chronic hepatitis patients would be used. Firstly, NK cell function pathways related genes in purified NK cells are tested and compared by RNA-seq and real-time PCR. Secondly, the differential genes’ function will be verified by siRNA technology. Thirdly, we co-culture healthy and chronic hepatitis patients’ NK cells transfected with the target genes and HBV infected HepaRG cells. Then the NK cell functions and the clearance abilities are detected. Finally, we test the gene expression of NK cells in liver with immunohistochemical technology. In conclusion, we intend to reveal the mechanisms why NK cells could suppress HBV in acute HIV infection, which will put forward more ideas of treating chronic HBV infection associated with NK cells.