| Â | Saline | Bleomycin |
|---|
WT | d21 | d7 | d14 | d21 |
|---|
AEI | 95.7 ± 4.87 | 79.47 ± 2.86 | 67.43 ± 1.76 | 58.41 ± 4.83 |
AEII | 4.3 ± 1.76 | 9.41 ± 1.86 | 8.66 ± 1.07 | 5.96 ± 1.12 |
NOX-/- | Â | Â | Â | Â |
AEI | 95.7 ± 3.87 | 66.21± | 58.23 ± 2.34 | 56.14 ± 3.97 |
AEII | 4.3 ± 1.12 | 4.89± | 5.43 ± 1.06 | 5.77 ± 0.56 |
DKO | Â | Â | Â | Â |
AEI | 95.7 ± 4.16 | 96.23 ± 4.75 | 96.44 ± 11.23 | 97.41 ± 3.87 |
AEII | 4.3 ± 0.56 | 4.5 ± 0.09 | 4.86 ± 1.94 | 4.96 ± 1.87 |
- Number of CD45- cells in LP counterstained with alveolar epithelial cell markers before and after bleomycin treatment.
- After the single intra-tracheal dose of bleomycin, animals were sacrificed on d7, d14, and d21 for the WT after bleo, and at D7 and d21 only for NOX-/- and DKO mice and wildtype saline treated mice. Cell number was quantitated by hemocytometer and corroborated by Z1 particle counter (Beckman Coulter). Number of cells represent data from 2 independent experiments ± SEM. N = 4/group. Cell subsets were quantified by FACS and morphologic differentiation under light microscope. Olympus BX41 at 40× magnification. Abbreviations used are: WT (wild type), NOX-/- (gp91phox knockout), gp91phox MMp12 double knockout (DKO). CD45+ cells were gated and macrophages (Gr1-F4/80hi), lymphocytes (CD3+ and B220+), and neutrophils (Gr-1hi F4/80-) were identified, their percentages analyzed by CellQuestPro and their total numbers calculated from the total number of cells obtained by lavage as described in Materials and methods. Data presented is pooled from 2 independent experiments ± SEM. Abbreviations used are: WS = Wildtype (WT) + saline (d7), WB7 = WT + post-bleomycin d7, WB14 = WT + post-bleomycin d14, WB21 = WT + post-bleomycin d21, NOXS = gp91phox-/- + saline d21, NOXB21 = gp91phox-/- + bleomycin d21, DKOS = gp91phox-MMP12 double knockout + saline (d21), DKOB21 = gp91phox-MMP12 double knockout + bleomycin (d21).
