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Table 1 Number of hematopoietic and non-hematopoietic cells in BALf and lung a week after bleomycin treatment - trends in cellular migration to BALf and lung post-bleomycin over time

From: Characterization of lung stem cell niches in a mouse model of bleomycin-induced fibrosis

A. Number of cells in BALF after a week of bleomycin treatment
  WT saline (day 7) WT bleomycin (day 7) NOX saline (d7) NOX bleomycin (day 7) DKO saline (day 7) DKO bleomycin (day 7)
Total cell count (×105/ml) 4.27 ± 0.184 28.3 ± 0.224 4.515 ± 0.223 48.91* ± 1.59 2.61 ± 0.16 8.42 ± 0.13
Macrophages (×10 5 /ml) 4.16 ± 0.57
(97.2%)
11.14 ± 4.08
(37.6%)
4.01 ± 0.31
(89.9%)
28.14* ± 6.83
(57.47%)
1.87 ± 0.64
(71.64%)
4.19 ± 3.18
(49.82%)
Lymphocytes (×10 5 /ml) 0.08 ± 0.01
(2.0%)
12.63 ± 2.49
(46.0%)
0.09 ± 0.02
(2.01%)
16.91* ± 3.65
(34.53%)
1.2 ± 0.01
(4.59%)
1.44 ± 0.66
(17.12%)
Neutrophils (×10 5 /ml) 0.03 ± 0.01
(0.7%)
4.5 ± 0.97
(16.3%)
0.11 ± 0.01
(0.02%)
3.91 ± 0.53
(7.98%)
0.63 ± 0.14
(24.13%)
2.78 ± 1.06
(33%)
B. Number of CD45 + cells in BALf before and after Bleomycin treatment
BALf         
10 5 /ml WS WB7 WB14 WB21 NOXS NOXB21 DKOS DKOB21
Total cells 4.27 ± 0.18 28.3 ± 0.22 10.32 ± 2.74 9.32 ± 1.74 4.51 ± 0.23 8.36 ± 1.32 4.55 ± 0.16 5.03 ± 1.94
Macrophage 4.16 ± 0.57 11.14 ± 4.08 8.91 ± 4.93 7.06 ± 3.92 3.86 ± 1.07 6.91 ± 1.96 3.91 ± 0.74 4.03 ± 0.43
Lymphocyte 0.08 ± 0.01 12.63 ± 2.49 1.76 ± 0.67 1.33 ± 0.67 1.32 ± 0.54 2.43 ± 0.43 1.44 ± 0.32 0.56 ± 0.21
PMN 0.03 ± 0.01 4.5 ± 0.97 0.41 ± 0.07 0.87 ± 0.32 0.67 ± 0.32 1.02 ± 0.22 0.67 ± 0.11 0.41 ± 0.13
C. Number of (CD45+) cells in lung parenchyma before and after bleomycin treatment
LP (d7)         
10 5 /m1 WS WB NOXS NOXB DKOS DKOB
Total cells 7.27 ± 0.07 38.3 ± 4.04 8.86 ± 0.08 54.19* ± 2.6 6.85 ± 0.27 9.56 ± 0.65
Macrophage 6.16 ± 0.57 22.14 ± 4.08 7.13 ± 2.43 31.38* ± 11.32 5.41 ± 1.43 6.36 ± 2.71
Lymphocyte 1.08 ± 0.01 9.63 ± 2.49 0.97 ± 0.12 21.41* ± 4.93 0.93 ± 0.21 2.47 ± 0.67
PMN 0.03 ± 0.01 7.5 ± 0.97 0.72 ± 0.04 1.34 ± 0.15 0.51 ± 0.03 0.73 ± 0.33
% WS WB NOXS NOXB DKOS DKOB
Macrophage 84.73 ± 1.86 57.80 ± 11.75 80.47 ± 12.96 57.96 ± 2.98 78.97 ± 3.76 66.52 ± 5.98
Lymphocyte 14.85 ± 2.84 25.14 ± 4.96 10.94 ± 1.95 39.50 ± 3.67 13.57 ± 3.98 25.83 ± 8.43
PMN 0.41 ± 0.04 19.58 ± 6.73 8.12 ± 2.07 2.47 ± 1.09 7.44 ± 1.97 7.63 ± 2.06
D. Number of CD45- cells in lunb parenchyma counterstained with alveolar epithelial cell markers before and after bleomycin treatment
  saline bleomycin
WT Day 21 Day 7 Day 14 Day 21
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-/-         
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         
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
  1. A. Number of cells in BALf after a week of bleomycin treatment. B. N umber of CD45+ cells in BALf before and on days 7, 14 and 21 after bleomycin treatment (one intratracheal dose of 0.074 U/ml in 40 ml volume). C. Number of (CD45+) cells in lung parenchyma (LP) before and after bleomycin treatment. D. 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 days 7, 14 and 21 for the WT after bleomycin, and at days 7 and 21 only for the NOX-/- and DKO mice and WS. 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. 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 were pooled from 2 independent experiments ± SEM. *P < 0.05 compared to WT post-bleomycin on corresponding time points). Abbreviations: WT (wild type), BALf, bronchoalveolar lavage fluid; NOX-/- (gp91phox knockout), gp91phox MMp12 double knockout (DKO); WS, WT + saline (day 7); WB7, WT + post-bleomycin day 7; WB14, WT + post-bleomycin day 14; WB21, WT + post-bleomycin day 21; NOXS, gp91phox-/- + saline day 21; NOXB21, gp91phox-/- + bleomycin day 21; DKOS, gp91phox-MMP12 double knockout + saline (day 21), DKOB21, gp91phox-MMP12 double knockout + bleomycin (day 21).