Fig. 8

A1 receptor signaling is required for PMN pulmonary influx. (A) Young, male mice were treated i.p with A1 inhibitor DPCPX and infected i.t with 5 × 10⁴ CFU S. pneumoniae TIGR4. At the indicated time points, lungs were harvested, digested, and stained for PMNs (CD45+, Ly6G + CD11b+) (A) and analyzed by flow cytometry. (B) Lungs of a separate set of treated and infected mice were harvested 6 hpi, homogenized and MPO levels were determined by ELISA. (C-D) Blood was harvested from infected and A1 treated mice (same mice as A) at the indicated time points and stained for PMNs and CD18 (C) or CXCR2 (D). (A, C and D) Data are pooled from 3 separate experiments with n = 6–12 mice per group. (A and D) Asterix indicate significance as determined by Kruskal Wallis followed by Dunn’s multiple comparison test. (B) Asterix indicate significance as determined by unpaired students t-test. (E) Young, male C57/B6 mice were treated i.p. with A1 inhibitor DPCPX or vehicle control (VC) and PMNs were isolated from the bone marrow. PMN migration across primary endothelial cells in response to S. pneumoniae TIGR4 infection was measured. Technical replicates are pooled from 4 separate experiments. Asterix indicates significance as determined by Mann-Whitney test