Figure 3

0∆NLS vaccination induces a greater number of plasmablasts and plasma cells expressing CD98 but no change in B regulatory cells in the spleen. C57BL/6 mice (n = 7/group) were injected in the footpad with the HSV-1 parental (GFP105) or live-attenuated HSV-1 0ΔNLS at 10⁵ plaque forming units (PFU) in 10 µl PBS. Mice were euthanized at day 7 and day 21 post-vaccination and the number of class-switched B cells was determined at day 7 in the PLN and day 21 in the spleen post-immunization. At day 7 and day 21 post-vaccination, the PLN and spleen were harvested and the number of plasmablasts (PBs) and plasma cells (PCs) expressing the CD98 marker were measured by flow cytometry. In addition, the number of CD19⁺CD21⁺CD23^highCD5⁺CD1d^high (regulatory B cell, Breg) along with marginal zone Breg cells (CD19⁺CD21⁺CD23^-CD5⁺CD1d^-) were evaluated by flow cytometry as well. Panels A and B present a gating strategy for the evaluation of PBs expressing CD98 (CD19+CD138+CD98+) and PCs expressing CD98 (B220lowCD138+CD98+), respectively. Panels C and D show the number of PBs and PCs expressing CD98 in the spleen at day 21 post-vaccination respectively. Panel E and F show the number of PBs and PCs expressing CD98 in the PLN at day 7 post-vaccination, respectively. Panel G presents a gating strategy to discriminate Breg and marginal zone Breg cells. Panels H and I show the number of Breg cells in the popliteal lymph node at day 7 and in the spleen at day 21 post-vaccination, respectively. Panels J and K present the number of marginal zone Breg cells in the popliteal lymph node at day 7 and in the spleen at day 21 post-vaccination, respectively. The results are presented as the mean + SEM, *p < .05 as determined by Student’s t-test.