Figure 1.
TACI deficiency leads to reduced MZ B cells. (A) Concentration of BAFF in the serum of Tnfrsf13b+/+ and Tnfrsf13b−/− mice. Bars show the mean. Each point represents one mouse. (B) Mixed bone marrow chimera generation. Bone marrow from Tnfrsf13b+/+ or Tnfrsf13b−/− littermates (expressing the CD45.2+ allele) was mixed 50:50 with bone marrow from WT CD45.1+ mice, then injected into sublethally irradiated Rag1−/− mice. B cells were analyzed at least 8 wk later. (C) Concentration of BAFF in the serum of WT:WT and KO:WT chimeras. Bars show the mean. Each point represents one mouse. (D) Stacked bar charts showing the percentages of CD45.2+ and CD45.1+ cells within the splenic MZ (CD93−CD1dhiIgMhi), FO (CD93−CD1dmedIgM+), and T2 (CD93+IgD+IgM+) B cell (B220+CD19+) populations in WT:WT and KO:WT chimeras. Cells were analyzed by flow cytometry using the gating strategy in Fig. S2 A. Bars show the mean ±95% CI. (E) Stacked bar chart showing the percentages of CD45.2+ and CD45.1+ cells within the splenic PC population (TCRβ−CD138+CD19lo) in WT:WT and KO:WT chimeras. Bars show the mean ±95% CI. (F) Representative flow cytometry plots showing the proportions of MZ and FO B cells within the CD45.2+ and CD45.1+ mature splenic B cell populations (CD93−B220+CD19+CD138−) in WT:WT and KO:WT chimeras. MZ and FO cells are gated via two different approaches. Numbers indicate the percentage of cells in that gate. (G and H) Proportions (G) and numbers (H) of MZ (CD1dhiIgMhi) and FO (CD1dmedIgM+) cells within the CD45.2+ and CD45.1+ mature splenic B cell populations (CD93−B220+CD19+CD138−) in WT:WT and KO:WT chimeras. Bars show the mean. Each point represents one mouse. Black: WT CD45.2+; red: TACI KO CD45.2+; gray: WT CD45.1+. Statistical tests: Welch’s t test (A and C), two-way ANOVA with Fisher’s LSD test (D and E), repeated-measures two-way ANOVA with Fisher’s LSD test (G and H). Numbers above graphs indicate P values where P ≤ 0.05, otherwise no value is shown. Sample numbers: 6 (A and D), 11 (C), 5 (E), 16 (G and H). Data are pooled from two experiments representative of four experiments (D), one out of two experiments (E), and pooled from six (A), five (C) and four (G and H) independent experiments. PC, plasma cell. Refer to the image caption for details. Panel A: A bar graph shows the concentration of BAFF (B cell Activating Factor) in the serum of Tnfrsf13b homozygous wild-type and Tnfrsf13b homozygous knockout mice. The y-axis represents the concentration of BAFF in nanograms per milliliter (ng/ml), and the x-axis shows the genotypes. Each point represents one mouse, and bars show the mean. Panel B: A schematic diagram illustrates the generation of mixed bone marrow chimeras. Bone marrow from Tnfrsf13b homozygous wild-type or Tnfrsf13b homozygous knockout littermates (expressing the CD45.2 positive allele) is mixed 50:50 with bone marrow from wild-type CD45.1 positive mice and then injected into sub-lethally irradiated Rag1 homozygous knockout mice. Panel C: A bar graph shows the concentration of BAFF in the serum of WT:WT (wild-type to wild-type) and KO:WT (knockout to wild-type) chimeras. The y-axis represents the concentration of BAFF in ng/ml, and the x-axis shows the chimera types. Each point represents one mouse, and bars show the mean. Panel D: Stacked bar charts show the percentages of CD45.2 positive and CD45.1 positive cells within the splenic marginal zone (MZ), follicular (FO), and transitional 2 (T2) B cell populations in WT:WT and KO:WT chimeras. The y-axis represents the percentage of cells, and the x-axis shows the chimera types. Bars show the mean 95 percent confidence interval (CI). Panel E: A stacked bar chart shows the percentages of CD45.2 positive and CD45.1 positive cells within the splenic plasma cell population in WT:WT and KO:WT chimeras. The y-axis represents the percentage of cells, and the x-axis shows the chimera types. Bars show the mean 95 percent confidence interval (CI). Panel F: Representative flow cytometry plots show the proportions of marginal zone (MZ) and follicular (FO) B cells within the CD45.2 positive and CD45.1 positive mature splenic B cell populations in WT:WT and KO:WT chimeras. The plots are gated using two different approaches, and numbers indicate the percentage of cells in each gate. Panel G: Bar graphs show the proportions of marginal zone (MZ) and follicular (FO) cells within the CD45.2 positive and CD45.1 positive mature splenic B cell populations in WT:WT and KO:WT chimeras. The y-axis represents the percentage of cells, and the x-axis shows the genotypes. Bars show the mean, and each point represents one mouse. Panel H: Bar graphs show the numbers of marginal zone (MZ) and follicular (FO) cells within the CD45.2 positive and CD45.1 positive mature splenic B cell populations in WT:WT and KO:WT chimeras. The y-axis represents the cell number normalized to wild-type CD45.2 positive cells, and the x-axis shows the genotypes. Bars show the mean, and each point represents one mouse.

TACI deficiency leads to reduced MZ B cells. (A) Concentration of BAFF in the serum of Tnfrsf13b+/+ and Tnfrsf13b−/− mice. Bars show the mean. Each point represents one mouse. (B) Mixed bone marrow chimera generation. Bone marrow from Tnfrsf13b+/+ or Tnfrsf13b−/− littermates (expressing the CD45.2+ allele) was mixed 50:50 with bone marrow from WT CD45.1+ mice, then injected into sublethally irradiated Rag1−/− mice. B cells were analyzed at least 8 wk later. (C) Concentration of BAFF in the serum of WT:WT and KO:WT chimeras. Bars show the mean. Each point represents one mouse. (D) Stacked bar charts showing the percentages of CD45.2+ and CD45.1+ cells within the splenic MZ (CD93−CD1dhiIgMhi), FO (CD93−CD1dmedIgM+), and T2 (CD93+IgD+IgM+) B cell (B220+CD19+) populations in WT:WT and KO:WT chimeras. Cells were analyzed by flow cytometry using the gating strategy in Fig. S2 A. Bars show the mean ±95% CI. (E) Stacked bar chart showing the percentages of CD45.2+ and CD45.1+ cells within the splenic PC population (TCRβ−CD138+CD19lo) in WT:WT and KO:WT chimeras. Bars show the mean ±95% CI. (F) Representative flow cytometry plots showing the proportions of MZ and FO B cells within the CD45.2+ and CD45.1+ mature splenic B cell populations (CD93−B220+CD19+CD138−) in WT:WT and KO:WT chimeras. MZ and FO cells are gated via two different approaches. Numbers indicate the percentage of cells in that gate. (G and H) Proportions (G) and numbers (H) of MZ (CD1dhiIgMhi) and FO (CD1dmedIgM+) cells within the CD45.2+ and CD45.1+ mature splenic B cell populations (CD93−B220+CD19+CD138−) in WT:WT and KO:WT chimeras. Bars show the mean. Each point represents one mouse. Black: WT CD45.2+; red: TACI KO CD45.2+; gray: WT CD45.1+. Statistical tests: Welch’s t test (A and C), two-way ANOVA with Fisher’s LSD test (D and E), repeated-measures two-way ANOVA with Fisher’s LSD test (G and H). Numbers above graphs indicate P values where P ≤ 0.05, otherwise no value is shown. Sample numbers: 6 (A and D), 11 (C), 5 (E), 16 (G and H). Data are pooled from two experiments representative of four experiments (D), one out of two experiments (E), and pooled from six (A), five (C) and four (G and H) independent experiments. PC, plasma cell.

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