Fig. 4.

Pik3cg-deficient mice recapitulate major features of human disease after exposure to natural microbiota through co-housing with pet-store mice. a mRNA expression of IL12b normalized to Hprt1 and Rpl13a in murine bone marrow-derived macrophages stimulated with LPS (100 ng/mL) and IFNγ (5 ng/mL) for 24 h. Data from two independent experiments (each with n = 2/group) are presented as mean ± SD. b Supernatant IL-12 concentration from murine bone marrow-derived macrophages stimulated with LPS (100 ng/mL) and IFNγ (5 or 20 ng/mL). Data are presented as mean ± SD and combined from three independent experiments. c Body weight of Pik3cg−/− and wild-type mice co-housed with pet-store mice. Data are representative of four independent experiments. d–f Frequency of Klrg1+ , CD62L+ , and CD44+ among CD8+ T cells from animals with or without exposure to pet-store mice at day 14. Data are presented as mean ± SD and are representative of four independent experiments (n = 3 for each non-co-housed group and n = 8 for each co-housed group). g Frequency of IL-10+ Foxp3+ cells among CD4 + T cells after PMA/ionomycin stimulation of splenocytes from WT (n = 2 for non-co-housed and n = 5 for co-housed) and Pik3cg−/− (n = 2 for non-co-housed and n = 6 for co-housed) after 56 days. Data are presented as mean ± SD. h Concentration of antibodies in serum from animals at 3–5 weeks after exposure to pet-store mice. Data are presented as mean ± SD, summarized from four independent experiments (n = 17–21 for each group). Statistical analysis was performed using unpaired T-test validated with Holm-Sidak method. i Total number of CD3 + T cells by immunohistochemistry per 200 μm section of small intestine from animals with or without exposure to pet-store mice at day 56. Data are presented as mean ± SD and representative of three independent experiments (n = 3 for non-co-housed and n = 7 for co-housed WT; n = 3 for non-co-housed and n = 6 for co-housed Pik3cg−/−). Statistical analysis was performed using two-tailed unpaired T-test in a–g and i