从HBV如何影响外周血中免疫细胞的天然免疫来探讨乙肝发生发展的机制，寻找靶向治疗的药物，是乙肝免疫治疗新的焦点。我们前期研究发现HBV在感染早期就能引起M2巨噬细胞的吞噬及多种细胞因子的表达。以此通过M2 mø蛋白磷酸化变化的质谱分析，得到Notch及NF-κB通路的潜在基因如RBPJ、IKKα。之后证实两者的表达受到抑制时，可以明显降低HBV诱导M2 mø吞噬、IL-6释放。为探索两通路间的交互作用，研究发现对IKKα的抑制，可降低RBPJ的表达。但具体机制需进一步探索。本项目采用shRNA处理的原代M2 mø与HBV共培养的条件下，研究M2 mø的免疫学功能变化。我们拟通过测序技术分析RBPJ、IKKα功能被阻断前后的mRNA水平变化，探讨两者交互作用。最后，对乙肝患者样本数据进行分析，研究Notch、NF-κB通路在临床特征中相关性，揭示HBV影响M2巨噬细胞天然免疫的具体作用机制。

It has been an important issue to investigate the innate immune response triggered by HBV infection, to explore the mechanism of hepatitis B infection and new efficient method to predict drug. Our previous study found that HBV could induce phagocytosis and expression of IL-1B, IL-8, IL-6, TNF-α and other cytokines in M2 macrophages. Through mass spectrometry analysis of M2 mø protein phosphorylation, RBPJ and IKKα are recognized as potential genes of Notch and NF-κB pathway in the immune process. And It has confirmed that the inhibition of RBPJ and IKKα expression significantly reduced internalization function and IL-6 release from the HBV associated M2 mø. Further studies have shown that the inhibition of IKKα, RBPJ expression can be significantly affected, but the mechanism is still unknown. In this project, shRNA was used to treat with M2 mø and then co-cultured with HBV. It has illustrated function of RBPJ and IKKα in M2 mø innate immunity, thereby our study such as M2 mø differentiation, maturation, antigenic abilities, M2 mø surface markers, phenotypes, M2 cytokine secretion levels, an HBV phagocytosis, etc. Besides, we intend to detect mRNA levels of gene library which before and after the blockade of RBPJ and Ikkα expression to further revealed the interaction between IKKα and RBPJ. Finally, the clinical data of hepatitis B patients will be analyzed and investigated to explore the clinical characteristics correlation between Notch and NF-κB pathway in innate immunity triggered by HBV infection. In this project, It has been elucidated new ways of immunotherapy for hepatitis B from a different perspective. It is also critical to study the mechanism of liver cirrhosis and hepatocellular carcinoma.