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. 2010 Feb 15;207(2):353–363. doi: 10.1084/jem.20091738

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© 2010 Linterman et al.

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Figure 4. — IL-21 contributes to affinity maturation but is dispensable for EM B cell formation. (A) Representative flow cytometric contour plots of donor CD45.1 SW_HEL cells from the spleens of Il21^+/+ or Il21^−/− recipient mice 10 d after adoptive transfer and immunization with HEL^3X SRBCs. Contour plots show IgG1⁺ cells binding with high (top oval gates) and low (bottom oval gates) affinity to HEL^3X (percentages are shown). (B) Bar graphs show HEL^{3X hi} IgG1⁺ CD45.1 cells (top oval gates in A) as a proportion of all HEL^3X-binding IgG1⁺ cells (square gates in A). This figure is representative of two independent experiments (n ≥ 3 mice per group. (C) Ratio of total Tfh (CXCR5^highPD-1^high CD4⁺) cells to total HEL-binding donor (CD45.1) GC B cells (Fas⁺GL-7⁺). Two independent experiments are represented in this plot. (D) Representative flow cytometric contour plots 10 d after adoptive transfer of SW_HEL B cells into Il21^+/+ or Il21^−/− mice and immunization with HEL^3X SRBCs gated on donor CD45.1 and HEL-binding cells (percentages are shown). The gates show GC cells (HEL^int binding) and EM B cells (HEL^hi binding). (E) Phenotype of GC B cells and EM cells gated as in D. (F and G) Total number of donor HEL^int-binding GC B cells per spleen (F) and the proportion of donor HEL^hi-binding EM B cells (G) in mice of the indicated genotypes gated as in D. Each symbol represents one mouse and tops of bars are drawn through the median values. Statistically significant differences are indicated (**, P ≤ 0.01).