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. 2017 May 24;8:595. doi: 10.3389/fimmu.2017.00595

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Copyright © 2017 Kratochvil, McKay, Kopycinski, Bishop, Hayes, Muir, Pinder, Cizmeci, King, Aldon, Wines, Hogarth, Chung, Kent, Held, Geldmacher, Dally, Santos, Cole, Gilmour, Fidler and Shattock.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of the memory B cell (mBC) response induced by sequential immunization with the human immunodeficiency virus-antigen CN54gp140. Frozen peripheral blood mononuclear cells (PBMCs) were thawed and expanded using human IL-2 and R848 for 4 days. Expanded cells were assayed for antigen (Ag)-specific and total IgG-secreting ASCs by ELISPOT assay at 0, 84, and 196 or 364 days, respectively. Each sample was measured in triplicate, averaged and plotted as Ag-specific per total mBC. (A) Percentage of Ag-specific mBC per total IgG-secreting mBC. Due to the limitation in availability of PBMC at least one data point is missing for the following participants: S001 (D0), S002 (D0), and S013 (D0 and D196). (B) Ag-specific spot forming units/well (indicated by lower indices) Ag-specific mBC and total-IgG-secreting mBC for one selected donor from each group for days 0, 84, and 196 or 364, respectively.