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目的接种乙肝疫苗是控制乙肝流行最经济有效的方法,通过对3次乙肝注射疫苗前后的B细胞库进行高通量测序分析,探讨注射乙肝疫苗后体液免疫的动态过程。方法将志愿者接种乙肝疫苗前后的外周血样进行密度梯度离心获得淋巴细胞并提取RNA,逆转录为cDNA后对B细胞受体H链的CDR3区序列进行高通量测序,以揭示注射乙肝疫苗前后B细胞应答的变化,分析人外周血B细胞受体CDR3的多样性以及序列变化与疫苗应答之间的关系。结果志愿者外周血B细胞受体CDR3的种类和数量在接种疫苗后都有所降低,并表现出其特有的免疫学特征,包括高频取用的V、D、J家族、V-J家族配对发生了改变,CDR3区氨基酸的取用和长度分布也发生显著变化;并且接种疫苗后,志愿者外周血中存在某些寡克隆异常增殖的情况。结论大部分志愿者在接种疫苗后,B细胞受体组库的多样性等发生特征性改变,并且出现某些疫苗接种相关的寡克隆。B细胞受体高通量测序技术可以从克隆水平上监控乙肝疫苗接种后抗体产生和表达的动态变化过程。
Objective Hepatitis B vaccine is the most economical and effective way to control the epidemic of hepatitis B. Through the high-throughput sequencing analysis of the B cell bank before and after 3 doses of hepatitis B vaccine, the dynamic process of humoral immunity after hepatitis B vaccine injection is discussed. Methods Peripheral blood samples from volunteers before and after hepatitis B vaccination were subjected to density gradient centrifugation to obtain lymphocytes and RNA was extracted. After reverse transcribed into cDNA, the CDR3 sequences of B cell receptor H chains were sequenced by high-throughput sequencing to reveal the pre- and post-injection hepatitis B vaccine B cell response changes in human peripheral blood B cell receptor CDR3 diversity and sequence changes and the relationship between vaccine response. Results The types and numbers of peripheral blood B cell receptor CDR3 in volunteers decreased after vaccination and showed their unique immunological characteristics including V, D, J family of high-frequency access and VJ family pairing Changes in access and length distribution of amino acids in CDR3 also changed significantly. After the vaccination, there were some oligoclonal abnormal proliferation in the peripheral blood of volunteers. Conclusions Most of the volunteers showed a characteristic change in the diversity of B cell receptor repertoire after vaccination, and some vaccine-related oligoclonal antibodies appeared. B cell receptor high-throughput sequencing can monitor the dynamic process of antibody production and expression after hepatitis B vaccination at the clonal level.