Clonal dissection of the human memory B‐cell repertoire following infection and vaccination

D Pinna, D Corti, D Jarrossay, F Sallusto… - European journal of …, 2009 - Wiley Online Library
D Pinna, D Corti, D Jarrossay, F Sallusto, A Lanzavecchia
European journal of immunology, 2009Wiley Online Library
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 …
Abstract
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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