Fig. 3: Kidney disease inhibits B cell response during T-dependent immunization.
From: The survival of B cells is compromised in kidney disease
A Schematic diagram of the experimental plan. AAN and control mice were i.p. immunized with 2.5% SRBC (n = 8) 4 days post-AAI injection. Seven days later, B total GC (liveB220+GL7+CD95+) [SRBC: Ctrl (8), AAI (8), AAII (8)], and C TFh (liveCD4+CD44+CXCR5+PD1+) cells were measured by flow cytometry [SRBC: Ctrl (8), AAI (8), AAII (8)]. AAN mice were either treated with probenecid (PRB + AAI) once on day 0 (relative to AAI injection) or left untreated. Control mice received probenecid only (PRB). Four days later, mice received NP-KLH immunization and assessed for D total GC B cells [PRB (9), AAI (7), PRB + AAI (9)], E TFh cells [PRB (9), AAI (10), PRB + AAI (9)], F NP-specific switched IgG1+ B cells (liveIgD−IgM−CD138− Gr1−B220+ NP+IgG1+) by flow cytometry [PRB (9), AAI (7), PRB + AAI (9)], and G serum NP4-specific IgG1 level by ELISA [PRB (8), AAI (6), PRB + AAI (8)]. H Diagram of the experimental design. WT mice were either subjected to 5/6 nephrectomy (5/6 Nx) (n = 7) or sham-operated (Ctrl) (n = 8). Two weeks after surgery, mice were immunized with NP-KLH and assessed for I total GC [Ctrl (8), 5/6 Nx (7)], J TFh cells [Ctrl (8), 5/6 Nx (7)], K NP-specific switched IgG1+ B cells [Ctrl (8), 5/6 Nx (6)], L serum NP4 and NP27-specific IgG1 levels [Ctrl (8), 5/6 Nx (7)], and M NP4/NP27- IgG1 ratio to measure affinity maturation [Ctrl (8), 5/6 Nx (7)]. Each dot represents individual mice and data are pooled from at least 2–3 independent experiments (B–G and I–M). Data expressed as Mean ± SD. Statistical analyses by One-way ANOVA (B–G) and two-sided t-test (I–M). B Ctrl vs. AAI **** P < 0.0001, AAI vs. AAII ****P < 0.0001. C Ctrl vs. AAI **P = 0.0027, AAI vs. AAII *P = 0.0160. D PRB vs. AAI **P = 0.0018, AAI vs. PRB + AAI *P = 0.0472. E PRB vs. AAI *P = 0.0112, AAI vs. PRB + AAI **P = 0.0029. F PRB vs. AAI **P = 0.0012, AAI vs. PRB + AAI *P = 0.0415. G PRB vs. AAI * P = 0.0222, AAI vs. PRB + AAI *P = 0.0484. I **P = 0.0096. J *P = 0.0278. K *P = 0.0295. L **P = 0.0028. M **P = 0.0049. ns: statistically not significant. Source data are provided as a Source Data file.
