HBV疫苗接种超高应答/低或无应答的效应、机制及初步应用研究

HBV疫苗接种者HBsAb高、低、无应答差异机制一直未完全阐明。课题组前期发现HBV疫苗接种者B细胞IgG & IgM H链CDR3组库存在多种个体化特征。本项目采用实验室率先建立CDR3组库HTS分析平台，筛选高、低、无应答HBV疫苗接种者，制备接种前、后PBMC样本。HTS分析B细胞IgG H & L链CDR3组库。系统解析B细胞CDR3组库多样性、克隆性等异同，BCR基因构成或取用缺失等个体化差异。同时，针对商品化乙肝免疫球蛋白生产存在的资源稀缺、价格受限、质量难控，高风险等问题，从B细胞CDR3组库数据中，筛选出与HBV疫苗高应答相关的IgG H & L链CDR3高频氨基酸序列，通过PCR扩增、载体构建、转染细胞、抗体纯化，获得HBV疫苗高应答相关单克隆抗体，并通过体、内外实验验证其功能。为HBV疫苗B细胞应答机制、高滴度HBsAb大量制备、HBV感染防治提供证据和新研究思路。

HBsAb in Hepatitis B vaccine recipients is high, poor or non-response, but the mechanism of response difference has not been fully clarified. Our previous works had found that there were many individual characteristics for the BCR heavy CDR3 repertoire of IgG&IgM in the HBV vaccine recipients. This project adopt HTS technology and analysis platform of BCR CDR3 repertoire to screen the HBV vaccination volunteers of high, poor or non-response. Prepare whole blood samples for PBMC isolation from the volunteers vaccinated against HBV. Illumina HTS IgG H & L CDR3 repertoire of B cells. Systematic comparative analysis each volunteer before and after injection of vaccine, and different volunteers having high, poor or non-response; similarities and differences of diversity and cloning about IgG H & L B cells CDR3; Individual differences in the composition or deletion of BCR genes. Meanwhile, in view of the problems existing in the production of commercial hepatitis B immunoglobulin, such as scarce resources, price constraint, quality difficult to control, high risk and so on. The high frequency amino acid sequence of IgG H & L chain CDR3 related to the high response of HBV vaccine was screened out from the data of B cell CDR3 group library. Through PCR amplification, vector construction, cell transfection and antibody purification, the monoclonal antibody related to high response of HBV vaccine was obtained, and its function was verified by in vivo and in vitro experiments. It can provide evidence and new research ideas for the mechanism of B cell response of HBV vaccine, the preparation of high titer HBsAb and the prevention and treatment of HBV infection.