PKCε在B细胞受体活化和免疫突触形成过程中作用的研究

Protein kinase C (PKC) is a family of kinases encoded by several genes with redundant and meanwhile distinct biological functions. One of such member, PKCε, is abundantly expressed in lymphocyte. However its roles in the formation of B cell immunological synapse (IS) and B cell activation are still unclear. Combine with high-speed high-resolution live cell single cell imaging and conventional biochemical analysis, we will explore the influence of PKCε to antigen-mediate BCR activation, microcluster growth and BCR pathway signaling transduction. By using DT40 signaling molecule KO cell library, we will decscrible the key molecules that determine the PKCε recruitment. The molecular mechanism of PKCε distription will also been addressed. What’s more, we will detect the effects of PKCε to the immune synapse formation in B cell immune synapse, and dig out the potential molecular mechanism. The roles of PKCε on primary and memory antibody responses will also be assessed by PKCε KO mice. These works will benefit our understanding to the regulation of B cell activation, which may help us to recognize abnormal activated B cell related diseases or vaccine design.

PKC是一类编码多种冗余但又均有独特生物功能的成员的激酶家族。其中PKCε在B淋巴细胞中表达丰富，但是其在B细胞激活和免疫突触形成过程中的功能未知。本申请为一年期的小额探索项目，我们将通过高速高分辨率活细胞单分子成像技术结合传统的生化细胞技术，重点阐述PKCε对抗原刺激的BCR微簇体的形成和成长过程的影响，对BCR信号通路下游信号分子激活的影响；我们还将借助DT40信号分子敲除细胞库分析PKCε对BCR信号通路分子依赖性的影响，并解释相应的PKCε自身分子特征对其招募后空间分布的影响；我们还将检测PKCε对BCR微簇体向免疫突触中心区域运输形成cSMAC的影响，并通过抑制剂处理进一步分析其可能的分子机制；此外，我们将借助PKCε敲除小鼠评估PKCε在初级和记忆性抗体反应中的作用。这些研究能够深化我们对B细胞激活调节机制的认识，可能在将来为B细胞活化异常导致的疾病的治疗和疫苗设计提供帮助。