Clonal dissection of the human memory B-cell repertoire following infection and vaccination.
Clonal dissection of the human memory B-cell repertoire following infection and vaccination.
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DOI：
10.1002/eji.200839129
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发表时间：
2009-05
影响因子：
5.4
通讯作者：
Lanzavecchia, Antonio
中科院分区：
文献类型：
--
作者：
Pinna, Debora;Corti, Davide;Jarrossay, David;Sallusto, Federica;Lanzavecchia, Antonio
对人类记忆B细胞库的分析具有重要的基础和实际意义。我们开发了一种简单的方法，使用R848和IL-2的组合选择性激活总新鲜或冷冻PBMC中的记忆B细胞。在这些条件下，30-40%的记忆B细胞在10天内产生平均产生200 ng IgG的克隆。该方法用于测量抗原特异性记忆B细胞的频率以及分泌的抗体的良好特异性、交叉反应性和中和活性。流感疫苗接种后，特异性B细胞显著扩增，在第14天达到总可克隆记忆B细胞的50%。特异性B细胞扩增也在未显示显著血清学应答的个体中检测到。动态变化和持久性的B细胞特异性的各种病原体记录在一系列PBMC样本收集了近二十年。这些结果揭示了记忆B细胞动力学的新方面，并提供了一个强大的工具来监测感染和接种疫苗后的免疫反应。
The analysis of the human memory B cell repertoire is of both fundamental and practical significance. We developed a simple method for the selective activation of memory B cells in total fresh or frozen PBMC using a combination of R848 and IL-2. In these conditions 30–40% of memory B cells generated clones producing on average 200 ng IgG in 10 days. This method was used to measure the frequency of antigen specific memory B cells as well as the fine specificity, crossreactivity and neutralizing activity of the secreted antibodies. Following influenza vaccination, specific B cells expanded dramatically, reaching up to 50% of total clonable memory B cells on day 14. Specific B cell expansions were detected also in individuals that did not show a significant serological response. Dynamic changes and persistence of B cells specific for a variety of pathogens were documented in serial PBMC samples collected over almost two decades. These results reveal novel aspects of memory B cell kinetics and provide a powerful tool to monitor immune responses following infection and vaccination.
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