Figure 1.
eYFP expression in CD19-Cre/eYFP mice.
A. CD19-Cre/eYFP mice were analyzed for eYFP expression in the B cell compartment of spleen, lymph nodes, Peyer's patches, peritoneal cavity and bone marrow. The shown histograms are gated on live lymphocytes and CD19. The filled histogram curve represents CD19-Cre/eYFP mice and the unfilled control mice. Percentages of eYFP+ CD19+ cells are shown in the histograms, upper number/percentage B-DTR mice, lower panel shows cells of control mice. B. To identify the CD19+ eYFP− B-cell fraction in bone marrow of CD19-Cre/eYFP mice, bone marrow cells were stained for AA4.1 and CD23.
Figure 2.
Diphtheria toxin mediated depletion of B cells in B-DTR mice.
The mice were treated once daily for 4 days with a dose of 25 ng/g body weight of diphtheria toxin. The decrease of B cells in comparison to T cells is shown in spleen, lymph nodes, bone marrow and peritoneal cavity; dot blots in the left panel and the corresponding bar charts in the right panel.
Figure 3.
Efficiency of B cell depletion.
A. Number of marginal zone, follicular, immature and mature B cells in spleen. B. Depletion of B cells in bone marrow, gated for B220+. C. Depletion of transitional B cells in spleen, gated for CD19+ cells. D. Depletion of mature and immature B cells in spleen, gated for CD19+ cells. E. Depletion of marginal zone and follicular B cells gated for CD19+ cells. F. Depletion of B1a and B2 B cells in peritoneal cavity, gated for CD19+. G. Depletion of resting B cells in peritoneal cavity gated for CD19+ the percentages of cells in the different shown B cell subpopulations are indicated.
Figure 4.
Total serum immunoglobulin levels after depletion of B cells.
B-DTR mice and control mice were injected daily four times with 25 ng/g body weight DT. Mice were bled just before DT injection and in weekly intervals thereafter. Shown are the total immunoglobulin levels at the indicated time points. The values are shown as average of groups of at least five mice.
Figure 5.
Plasma cells identified with Syndecan-1 in cytospins.
Bone marrow cells from NP-CG immunized and boosted cells were analyzed for plasma cells. Plasma cells are shown in red with a surface staining of syndecan-1, the remaining cell population was counter stained with Hoechst (blue). The bar chart represents plasma cells found in the regarding groups (n = 5). The pictures show representative sections of the cytospins.