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, cross-reactivity 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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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3864550PMC
http://dx.doi.org/10.1002/eji.200839129DOI Listing

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