B cell receptor revision diminishes the autoreactive B cell response after antigen activation in mice
J. Clin. Invest. Ying-Hua Wang, et al. 118:2896 doi:10.1172/JCI35618 [Go to this article.]

Figure 3
DWEYS-MAP induced RAG expression requires IL-7R signaling. (A) Flow cytometry analysis of IL-7R expression on Tet⁺B220^lo (solid line), Tet⁺B220^hi (dotted line), and Tet^–B220^hi (line over shaded area) subsets. Splenic cells were prepared on day 16 from DWEYS-MAP–immunized mice. The experiment was repeated twice with 5 mice at each time. The Tet⁺B220^lo cells display the highest increase in the level of IL-7R protein. (B) qPCR of Il7r on the specified B cell subsets. Cells from DWEYS-MAP–immunized mice were sorted by flow cytometry on day 16. Data (mean ± SEM) represent 1 of 3 independent experiments. Tet⁺B220^lo cells show the greatest increase in Il7r mRNA. (C) Histological staining of spleen sections for B220 (red) and RAG2 (green). Mice were immunized with DWEYS-MAP on day 0, boosted on day 7, and treated with anti–IL-7R–blocking antibody or with isotype control on days 8, 11, and 14. Spleens were harvested on day 16. Three mice were included in each group. A minimum of 6 pictures were taken for each spleen. The experiment was repeated twice. Spleens from mice treated with anti–IL-7R antibody showed decreased RAG2 expression (green). Follicles are identified by B220 (red). Original magnification: ×50. (D) qPCR analysis of Rag2 in Tet⁺B220^lo cells from DWEYS-MAP–immunized mice treated with anti–IL-7R Ab or with isotype control. The procedure for immunization and administration of antibodies was described in C. Cell sorting was done on day 16, following immunization. Data (mean ± SEM) are representative of 3 independent experiments. Rag2 mRNA is undetectable in mice treated with anti-IL7R antibody. For all these experiments, the mice were immunized with DWEYS-MAP in CFA on day 0 and boosted with DWEYS-MAP in incomplete Freund adjuvant on day 7.