Fig. 4

Repetitive ODE exposure induces airway and lung compartment B1 and B2 populations. WT and BCR KO mice were repetitively exposed to saline or ODE daily for 3 weeks whereupon BALF and dissociated tissue cells were processed and analyzed by flow cytometry. Lymphocytes identified by CD45⁺ leukocytes excluding debris and characteristic FSC and SSC properties of lymphocytes followed by staining for CD19, CD11b, and CD5. A representative contour plot of a BALF cells and c lung tissue cells demonstrating gating strategy for CD19⁺ B cells, B2 B cells (CD19⁺CD11b⁻), B1 B cells (CD19⁺CD11b⁺), and B1a B cells (CD19⁺CD11b⁺CD5⁺) from WT saline and ODE treated animals is shown. Numbers of B, B2, B1, and B1a cells were calculated by multiplying the percentage of cells in respective gate (% of CD45+ cells, as analyzed by FACS) multiplied by respective total cells (see Fig. 3) for each mouse. Bar graphs depict means with standard error bars of b BALF and d lung tissue cells of 6 mice/group. There were no B cells in the BCR KO mice. Statistical difference (*p < 0.05; **p < 0.01) versus saline