FIGURE 7.

CLP EVs promote peritoneal leukocyte recruitment in vivo. WT, TLR7^−/−, and MyD88^−/− male mice were administered i.p. saline, 300 μg of sham EVs, or CLP EVs. Twenty hours later, the peritoneal lavage was harvested, and the peritoneal cells were analyzed with flow cytometry. (A) Gating strategy for flow cytometry. F4/80^high–Alexa Fluro 647 were determined as resident Mϕ and Ly-6G⁺-Percp-Cy5.5 were determined as NE. (B) Representative flow cytometry of gated F4/80^high and Ly-6G⁺ in peritoneal cells of WT mice i.p. injected with saline, sham EVs, or CLP EVs. (C) CLP EVs reduce resident Mϕs and promote NEmigration into the peritoneal cavity as compared with sham EVs (n = 6). (D and E) TLR7 deficiency has no impact on CLP EV–induced Mϕ reduction and NE increase in the peritoneal cavity (n = 5–6). (F and G) MyD88-deficient mice had reduced NE migration to the peritoneal cavity after sham EV and CLP EV injection as compared with WT mice (n = 3–4). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.