Figure 1. Anergic B cells acquire an actived phenotype during γHV68 infection but do not break tolerance.

A–D) Ars/A1-C57BL/6 chimeric mice were infected with γHV68 or sham-infected (n=5/group). A) On the indicated days post infection the cell-surface phenotype was determined of the Ars/A1 B cells (B220⁺IgM^a+) and the C57BL/6 B cells (B220⁺IgM^a−). Data from infected mice is presented as thick lines, data from uninfected control mice as filled histograms, representative histograms are shown. B) Ars/A1 derived serum antibody (IgM^a anti-Ars) at day 12 post infection (OD405 at 1:40 serum dilution). C) At day 13 post infection spleen cells from chimeric mice were stained with anti-IgM^a/B220 or anti-IgM^b/B220, loaded with indo-1 and stimulated with 5μg F(Ab)₂ anti-IgM. Changes in intracellular calcium in Ars/A1 B cells was determined by gating on B220⁺IgM^b− events, C57BL/6 (WT) by gating B220⁺IgM^a− events. D) chimeric mice were immunized i.p. with 100μg OVA-Ars/FIA 3 days post infection, 13 days later serum was collected and the IgM^a anti-Ars response was determined. Error bars show SEM, * p<0.05. All data is representative of at least 2 independent experiments.