Fig. 2

B cell-intrinsic impact of the DOCK11 deficiency on recall responses. A, Comparable formation of NP-specific IgG1⁺ non-GC B cells (B220⁺CD38⁺CD95⁻NP⁺IgG1⁺) in the spleen of the indicated strains 28 d post-immunization with alum-precipitated NP-CGG, as measured by flow cytometry. Representative pseudo-color plots are gated on B220⁺CD38⁺CD95⁻ cells. Numbers show percentages of cells in each gate. Detailed gating strategies are shown in Fig. S1. B, Numbers of NP-specific IgG1⁺ non-GC B cells in (A). Each point represents an individual mouse. Bars represent geometric means. Data are pooled from two independent experiments, using five or more mice per experimental group. C, Experimental outline to examine the impact of the DOCK11 deficiency on recall responses of antigen-experienced IgG1⁺ non-GC B cells. NP-specific IgG1⁺ non-GC B cells were isolated from spleens of B1–8 IgH-carrying mice (WT) or DOCK11-deficient counterparts (KO) 28 d post-immunization with alum-precipitated NP-CGG. These cells were then transferred into congenic C57BL/6 recipients (WT) immunized with alum-precipitated CGG 28 d before, followed by secondary immunization with NP-CGG and subsequent analyses 7 d later. D, Numbers of NP-specific IgG1 antibody-secreting cells (ASCs) as measured by an ELISPOT assay in the spleen of recipients given transfer or not (−) of DOCK11-sufficient (WT) or deficient (KO) NP-specific IgG1⁺ non-GC B cells, followed by secondary immunization with NP-CGG. Each point represents an individual recipient. Bars represent geometric means. Data are pooled from three independent experiments, using eight or more recipients per experimental group. The values of anti-NP IgG1⁺ASC/10⁵ splenic cells ± SE are 0.9 ± 0.2 (−), 2.7 ± 0.7 (WT) and 2.7 ± 0.4 (KO). E, Serum levels of NP-specific IgG1 in (D), as measured by an ELISA. **P < 0.01, *P < 0.05 (Tukey’s test)