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. 2007 Jun 11;204(6):1383–1393. doi: 10.1084/jem.20061205

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Copyright © 2007, The Rockefeller University Press

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Figure 6. — Examination of the response of NKCC1^−/− and control animals in a sterile model of acute lung inflammation. The total number of cells in the BALF was enumerated (A), and populations of leukocytes were determined based on morphological criteria (B). The total number of cells in the BAL was significantly increased in LPS-challenged NKCC1^−/− mice (P = 0.00007; n = 11) compared with the LPS-challenged NKCC1^+/+ mice (n = 12). Differential cell counts showed that this increase was the result primarily of increased numbers of neutrophils (P = 0.0001) although a small but significant increase in the number of macrophages was also observed (P = 0.004) in NKCC1-deficient mice compared with wild-type controls. To determine whether this difference represented a change in sequestration of neutrophils in the lung or specifically the movement of the cells into the airway, lungs were harvested from similarly treated animals, and the level of MPO in the lung was determined (C). MPO levels in the lungs of NKCC1^−/− mice (n = 11) were higher than in the wild-type controls (n = 12), but the increased levels did not reach statistical significance. To directly visualize differences in the location of inflammatory cells in the two mouse lines, lungs were fixed and sections were prepared from LPS and saline-treated animals 12 h after exposure. Images similar to those shown in E were scanned and analyzed using Image Scope viewing software, and cells present in the airways were enumerated in a blinded fashion. The number of neutrophils present in the airways of the NKCC1^−/− lungs (n = 4) was higher than in the control animals (n = 4) (D). All values are expressed as mean ± SEM. *, P < 0.005 and **, P < 0.0005. Bars: (top) 50 μm; (bottom) 100 μm.