Fig. 2.

Microglia depletion by a Csf1r antagonist in EAU does not alter the systemic immune response to the immunized peptide. C57BL/6 mice were fed the Csf1r antagonist PLX5622 (PLX) or a control diet 7 d before IRBP immunization, and animals were immunized on day 0. (A–E) Animals were evaluated for a systemic immune response against the IRBP-p. (A) Delayed hypersensitivity, as determined by ear swelling, was evaluated on day 21. Mice were injected intradermally with IRBP-p into the pinna of one ear on day 19. Ear swelling was measured after 48 h using a micrometer (n = 6–7 mice per group). (B–E) Cell proliferation was evaluated by using the cells isolated from LNs and spleens (SPs) on days 14 and 21. The cells were cultured in triplicate for 3 d in the presence of IRBP-p (10 μg), Con A (1 μg), or medium only (n = 5 mice per group). During the last 4 h before the 72-h end-point culture, the Cell Counting Kit-8 was added to each well. At 72 h, viable cell numbers in each well were measured as the absorbance (450 nm) of reduced WST-8. (F and G) Weights of the draining LNs (n = 5 mice per group) and SPs (n = 5 mice per group) were measured on day 21. Data were analyzed by one-way ANOVA, followed by Tukey’s multiple comparison test. Data are expressed as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. n.s., not significant.