Figure 4. LysM-cre Socs3^fl/fl mice recruit T and NK cells with impaired IFN-γ production.

T and NK cells traffic to the peritoneal cavity in appropriate numbers by day 4 following infection of LysM-cre Socs3^fl/fl mice yet exhibit a less activated phenotype with decreased IFN-γ production. Socs3^fl/fl (n=5) and LysM-cre Socs3^fl/fl (n=5) were infected i.p. with 10⁴ Pru parasites and sacrificed at days 4 and 8 post-infection. (A) The total number of NK cells (B220⁻, CD3⁻, NK1.1⁺), CD4⁺ (B220⁻, CD3⁺, CD4⁺), and CD8⁺ (B220⁻, CD3⁺, CD8⁺) T cells were determined by flow cytometry. (A,B) CD4⁺ T cells were measured for the surface markers CD62L and CD44 and (C) quantified for the number of activated (CD44^hi, CD62L^low) and naïve (CD44^low, CD62L^hi) cells per mouse. (1×10⁶ (D,E) For intracellular cytokine staining, PECs were incubated with BFA, Golgi Stop, PMA, and ionomycin for 3 hrs and analyzed for IFN-γ production following surface staining for NK cells, and CD4⁺ and CD8⁺ T cells. (F,G) 1×10⁶ splenocytes were plated from extracted spleens of infected mice and stimulated with media alone or 50 µg/mL STAg. Concentrations of IL-12p40 and IFN-γ were measured by ELISA following 24 hrs of stimulation. One-Way ANOVA followed by student’s t test was used to determine statistical significance. Error bars +/− SD. Results are representative of two independent experiments.