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. 2020 Oct 29;218(2):e20191393. doi: 10.1084/jem.20191393

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© 2020 Müller-Winkler et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 5. — BAFF depletion results in the loss of MBCs. (A) WT mice were immunized with NP-CGG in alum or alum alone on day 0, followed by anti-CD40L or isotype control antibody injections on days 55, 57, and 59. Anti-BAFF or isotype control antibody was injected on days 60 and 64. (B) Mean (±SEM) BAFF levels in serum of mice immunized with NP-CGG in alum 21 d after anti-BAFF (n = 11) or isotype control antibody (n = 9) injection. Each dot represents one mouse. (C–E) Mean (±SEM) numbers of B cell populations in spleen (C), bone marrow in two legs (D), or blood (E) of mice immunized with NP-CGG in alum (+; n = 10 for anti-BAFF and n = 12 for isotype control antibody in spleen; n = 11 for anti-BAFF and n = 14 for isotype control antibody in bone marrow; and n = 15 for anti-BAFF and n = 13 for isotype control antibody in blood) or alum alone (−; n = 5) as described in A, 21 d after last anti-BAFF or isotype control antibody injection. Splenic T cells (CD3⁺CD138⁻B220⁻), FO (FO B, CD3⁻CD138⁻B220⁺Fas⁻AA4.1⁻PD-L2⁻IgM^loCD23⁺), MZ (MZ B, CD3⁻CD138⁻B220⁺Fas⁻AA4.1⁻PD-L2⁻IgM^hiCD23⁻), and NP⁺ IgG1⁺ and IgM⁺ MBCs and NP⁻ IgG1⁺ MBCs (CD3⁻CD138⁻B220⁺Fas⁻AA4.1⁻PD-L2⁺), bone marrow mature B cells (CD138⁻B220^hiIgM⁺), PCs (B220⁻CD138^hi), and NP⁻ IgG1⁺ MBCs and NP⁺ IgM⁺ and IgG1⁺ MBCs (CD138⁻B220^hiPD-L2⁺), and blood T cells (CD3⁺B220⁻), B cells (CD3⁻B220⁺CD138⁻), and IgM⁺, IgG1⁺, or IgG2b⁺ MBCs (CD3⁻B220⁺CD138⁻PD-L2⁺) were analyzed using the gating strategy shown in Fig. S4 B. Each dot represents one mouse. (F) WT mice were infected i.n. with X31 influenza or given PBS as a control on day 0, followed by anti-CD40L or isotype control antibody injections on days 52, 54, and 56. Anti-BAFF or isotype control antibody was injected on days 57 and 61. (G) Mean (±SEM) numbers of cell populations in lungs of mice infected with X31 influenza (+; n = 16 for anti-BAFF and n = 14 for isotype control antibody) or given PBS as a control (−; n = 4) 19 d after last anti-BAFF or isotype control antibody injection. Pulmonary T cells (CD3⁺CD19⁻), total B cells (CD3⁻CD138⁻CD19⁺), and HA⁺ IgM⁺, IgG1⁺, IgG2b⁺, and IgG2c⁺/IgA⁺ and HA⁻ IgG1⁺, IgG2b⁺, and IgG2c⁺/IgA⁺ MBCs (CD3⁻CD138⁻CD19⁺PD-L2⁺) were analyzed using the gating strategy shown in Fig. S4 D. Data pooled from two (C–E) and three (G) independent experiments. Mann-Whitney test was used for statistical analysis. *, 0.01 < P < 0.05; **, 0.001 < P < 0.01; ***, 0.0001 < P < 0.001; ****, P < 0.0001.