Disruption of PTEN enhances alveolar macrophage accumulation and elevates proinflamatory cytokine/chemokine production in the inflamed lungs. Mice were intratracheally instilled with 106 CFU of E coli and killed at indicated time points. BALF were collected using ice-cold PBS/15 mM EDTA (ethylenediaminetetraacetic acid; 1 mL ×10). (A) Differential cell counts were conducted on cytospin preparations stained with a modified Wright-Giemsa stain (Volu-Sol). Macrophages were identified morphologically. BALF total cell numbers were counted using a hemocytometer. Macrophage population was calculated according to its percentage. (B) Total numbers of leukocytes, neutrophils, resident macrophages, and inflammatory macrophages in BALF were determined by FACS analysis. (C) BALF chemokine and cytokine levels were determined using enzyme-linked immunosorbent assay (ELISA) kits. (D) LPS-induced chemokine and cytokine release from purified primary alveolar macrophages. Alveolar macrophages were collected from unchallenged WT and PTEN KO mice and stimulated with LPS (1 μg/mL) for up to 24 hours. Supernatants were harvested and the chemokine/cytokine levels were determined using ELISA kits. Data are presented as mean (± SD; n ≥ 3 mice in each group). *P < .05, **P < .01 versus WT.
