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Fig. 3 | Immunity & Ageing

Fig. 3

From: The immunosenescence-related factor DOCK11 is involved in secondary immune responses of B cells

Fig. 3

B cell-extrinsic impact of the DOCK11 deficiency on recall responses. A, Experimental outline to examine whether the lack of DOCK11 in B cells affects the secondary immune responses in a B cell-extrinsic manner. NP-specific IgG1+ non-GC B cells were isolated from spleens of B1–8 IgH-carrying mice 28 d post-immunization with alum-precipitated NP-CGG. These cells were transferred into congenic Cd19-Cre mice or DOCK11-deficient counterparts immunized with alum-precipitated CGG 28 d before, followed by secondary immunization with NP-CGG and subsequent analyses 7 d later. B, Numbers of NP-specific IgG1 antibody-secreting cells (ASCs) as measured by an ELISPOT assay in the spleen of the indicated recipients given transfer of 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 five or more recipients per experimental group. The values of anti-NP IgG1+ASC/105 splenic cells ± SE in Cd19-Cre control group and Dock11flCd19-Cre group are 2.8 ± 0.5 and 1.6 ± 0.2, respectively. C, Serum levels of NP-specific IgG1 in (B), as measured by an ELISA. D, Experimental outline to examine whether the impaired recall responses of antigen-experienced B cells in the DOCK11-deficient recipients would be recovered by adoptive transfer of cognate CD4+ T cells. NP-specific IgG1+ non-GC B cells were isolated from spleens of B1–8 IgH-carrying mice 28 d post-immunization with alum-precipitated NP-OVA. CD4+ T cells were isolated from spleens of naïve C57BL/6 mice or OT-II mice 28 d post-immunization with alum-precipitated OVA. NP-specific IgG1+ non-GC B cells were transferred in combinations with either naïve CD4+ T cells or OVA-primed OT-II T cells into congenic Dock11flCd19-Cre mice immunized with alum-precipitated OVA 28 d before, followed by secondary immunization with NP-OVA and subsequent analyses 7 d later. E, Numbers of NP-specific IgG1 ASCs as measured by an ELISPOT assay in the spleen of the recipients given transfer of NP-specific IgG1+ non-GC B cells and the indicated CD4+ T cells. Each point represents an individual recipient. Bars represent geometric means. Data are pooled from two independent experiments, using four recipients per experimental group. The values of anti-NP IgG1+ASC/105 splenic cells ± SE in naïve CD4+T control group and OVA-primed OT-II T group are 0.9 ± 0.2 and 2.6 ± 0.6, respectively. F, Serum levels of NP-specific IgG1 in (E), as measured by an ELISA. **P < 0.01, *P < 0.05 (Welch’s t-test)

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