Eventual loss of HA-specific ASCs and HA-specific serum IgG after depletion of HA-specific MBC.

<p>8–10 wk old female hCD20tg and hCD20tg-negative littermates were infected with Influenza A/PR/8/34, and treated with anti-hCD20 mAbs as described in the <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003843#s4" target="_blank">Materials and Methods</a>. <b>A.</b> HA-specific IgG MBCs in spleens from hCD20tg (○) and hCD20tg-negative (•) just before treatment and 1 day, 42 days, 70 days and 150 days post-treatment [hCD20tg: n = 5(d0), 5(d1), 3(d42) and 3(d150); hCD20tg-negative littermates: n = 4(d0), 2(d1), 2(d42) and 3(d150)]. The data are pooled from 3 independent experiments. <b>B–D.</b> Femur pairs were obtained from mice and HA-specific and total IgG bone marrow ASCs were quantified using ELISpot in hCD20tg (○) and hCD20tg-negative (•) mice prior to treatment and 1 day, 42 days, 70 days and 150 days post-treatment. [hCD20tg: n = 7(d0), 5(d1), 5(d42) and 3(d150); hCD20tg-negative littermates: n = 5(d0), 2(d1), 2(d42) and 3(d150)]. The graphs show number of HA-specific ASC <b>B,</b> per 10⁶ bone marrow cells and <b>C,</b> per femur pair; <b>D,</b> number of total IgG ASC per femur pair; and <b>E,</b> numbers of HA-specific IgG ASC expressed as a ratio to total IgG ASC. The data are pooled from 3 independent experiments. <b>F.</b> Serum was obtained and HA-specific serum IgG were quantified in hCD20tg (○) and hCD20tg-negative (•) just before treatment, and 42, 70, 90 and 150 days post-treatment by ELISA [hCD20tg: n = 13(d0), 4(d42), 8(d90) and 8(d150); hCD20tg-negative littermates: n = 13(d0), 3(d42), 8(d90) and 8(d150)]. The data were obtained from 2 experiments. In figures <b>A–F</b>, error bars indicate the median ± error and statistical values were calculated using a two-tailed Mann-Whitney test. Non-significant values (P>0.05) are not indicated on the graphs.