Panel A shows box plots comparing germinal center B220 positive cells and lymph node germinal center cell counts in unimmunized, wildtype, and Sorl1 knockout mice. Panel B shows box plots comparing nitrophenyl-specific germinal center B cells and nitrophenyl positive germinal center cell counts between wildtype and Sorl1 knockout mice. Panel C shows a box plot comparing T follicular helper cells as a percentage of T-cell receptor positive cells. Panel D shows box plots comparing nitrophenyl-specific memory B cells and memory B-cell counts. Panel E shows box plots comparing plasma cells as a percentage of B220 positive cells and plasma cell absolute counts. Panel F shows a box plot comparing the percentage of mCherry positive germinal center cells normalized to the initial infection. Panel G shows a box plot comparing the percentage of mCherry positive plasma cells normalized to the initial infection. Panel H shows a box plot comparing the percentage of mCherry positive Immunoglobulin G positive plasma cells normalized to the initial infection. Panel I shows box plots comparing low-affinity nitrophenyl-specific Immunoglobulin M levels over time. Panel J shows box plots comparing high-affinity nitrophenyl-specific Immunoglobulin M levels over time. Panel K shows a box plot comparing the nitrophenyl 4 to nitrophenyl 36 ratio on Days 7 and 14. Panel L shows box plots comparing low-affinity nitrophenyl-specific Immunoglobulin G1 levels over time. Panel M shows box plots comparing high-affinity nitrophenyl-specific Immunoglobulin G1 levels over time. Panel N shows a box plot comparing the nitrophenyl 8 to nitrophenyl 36 ratio on Days 7 and 14.
SorLA contributes to humoral immune responses in vivo. (A) Percentage and absolute counts of lymph node GC B cell populations 14 days following immunization with NP-CGG in WT, Sorl1−/−, and unimmunized WT mice. P value shown calculated by Mann–Whitney U test, n = 9–12 mice from three experiments. (B) Percentage and absolute counts of lymph node NP-specific GC B cell populations 14 days following immunization with NP-CGG in WT, Sorl1−/−, and unimmunized WT mice. n = 4–5 mice from two experiments. (C) Lymph node Tfh cell populations 14 days following immunization with NP-CGG in WT, Sorl1−/−, and unimmunized WT mice. n = 5 mice from one experiment. (D) Percentage and absolute counts of lymph node memory B cell populations 14 days following immunization with NP-CGG in WT, Sorl1−/−, and unimmunized WT mice. n = 7–8 mice from two experiments. (E) Percentage and absolute counts of lymph node plasma cell populations 14 days following immunization with NP-CGG in WT, Sorl1−/−, and unimmunized WT mice. n = 8–10 mice from two experiments. (F) Development of GC cell populations in spleen of Cd4- or Sorl1-targeted bone marrow chimeras at 14 days after immunization. % mCherry+ cells is normalized to the initial infection (% mCherry+) of HSC for each sgRNA per experiment. n = 7 mice from two independent experiments. (G) Development of plasma cell populations in spleen of Cd4- or Sorl1-targeted bone marrow chimeras at 14 days after immunization. % mCherry+ cells is normalized to the initial infection (% mCherry+) of HSC for each sgRNA per experiment. n = 7 mice from two independent experiments. (H) mCherry expression in class-switched IgG+ plasma cell populations in spleen of Cd4- or Sorl1-targeted bone marrow chimeras at 14 days after immunization. % mCherry+ cells is normalized to the initial infection (% mCherry+) of HSC for each sgRNA per experiment. n = 7 mice from two independent experiments. (I) Low-affinity NP-specific IgM antibody production, quantified by NP36-binding ELISA. n = 19–20 mice from four independent experiments. (J) High-affinity NP-specific IgM antibody production, quantified by NP(8) binding ELISA. n = 19–20 mice from four independent experiments. (K) Ratio of IgM binding to NP(8) and NP(36). n = 19–20 mice from four independent experiments. (L) Low-affinity NP-specific IgG1 antibody production, quantified by NP36-binding ELISA. n = 19–20 mice from four independent experiments. (M) High-affinity NP-specific IgG1 antibody production, quantified by NP(8)-binding ELISA. n = 19–20 mice from four independent experiments. (N) Ratio of IgG1 binding to NP(8) and NP(36). n = 19–20 mice from four independent experiments. Data are shown as box and whisker plots, where the line indicates the median, the box represents the interquartile range, and whiskers denote the minimum and maximum values. P values shown in A–H are calculated by Mann–Whitney U test. P values shown in I–N are calculated by two-way ANOVA with multiple comparisons.