Article Figures & Data
Figures
- FIGURE 1.
Cellular composition in BALF. Number of cells in BALF harvested from TNF+/+ (open bars) and TNF−/− (filled bars) mice after BLM instillation (a) or sterile PBS treatment (b). BALF on day 0 was harvested from nontreated mice. Data represent mean ± SEM (n = 5 for each group). ∗, p < 0.01. ∗∗, Few of neutrophils and lymphocytes were observed in nontreated mice.
- FIGURE 2.
Cellular composition of lung cells. Number of cells in lungs isolated from TNF+/+ (open bars) and TNF−/− (filled bars) mice on 21 days after BLM instillation (a) or sterile PBS treatment and none (b). Data represent mean ± SEM (n = 5 for each group). ∗, p < 0.01.
- FIGURE 3.
Histological analysis of lung damage induced by BLM. Lungs were collected on days 14 (a, b) and 75 (c, d) after BLM injection. Lungs from TNF+/+ (a, c) and TNF−/− (b, d) mice. The paraffin sections were H&E stained and photographed at ×40 magnification. Results are representative of four mice in each group.
- FIGURE 4.
Hydroxyproline content in lungs after BLM instillation. Lungs were collected on day 21 after BLM or sterile PBS injection. Hyroxyproline content was measured in TNF+/+ (open bars) and TNF−/− (filled bars) mice. Data represent mean ± SEM (n = 5 for each group).
- FIGURE 5.
Production of TNF in lungs and expression of TNFRs after BLM instillation. a, TNF production in lungs was determined by ELISA. TNF+/+ (•) and TNF−/− mice (○). Data represent mean ± SEM (n = 5 for each group). b and c, Flow cytometric analysis of expression of TNFRs on inflammatory cells in BALF. Numbers represent percentage of TNFR-positive cells. Results are representative of five mice.
- FIGURE 6.
Apoptotic cells in the bronchoalveolar space after BLM instillation. a, Detection of apoptotic cells by flow cytometric analysis. BAL was performed on day 14 after BLM. Numbers represent percentage of annexin V- and/or propidium iodide-positive cells. Results are representative of five mice. b, Annexin V-positive cells were counted as apoptotic cells. TNF+/+ mice (open bars) and TNF−/− (filled bars). Data represent mean ± SEM (n = 5 for each group), except for those obtained on day 28, which represent pooled inflammatory cells harvested from five mice. ∗, p < 0.05.
- FIGURE 7.
Effects of airway challenge with rTNF on induction of apoptosis of inflammatory cells. a, Number of inflammatory cells in the BALF harvested from TNF−/− mice challenged via airways with murine rTNF (filled bars) or PBS alone (open bars) after BLM. The concentration of murine rTNF was 250 ng/ml in PBS. BAL was performed 48 h after airway challenge with TNF. Data represent mean ± SEM (n = 5 for each group). ∗, p < 0.01. b, Number of inflammatory cells in BALF harvested from TNF−/− mice challenged with murine (filled bars) or human rTNF (open bars) on day 14 after BLM instillation. TNF concentration was 250 ng/ml in PBS. BAL was performed sequentially after airway challenge with TNF. Data represent mean ± SEM (n = 5 for each group). ∗∗, p < 0.05. ND, not done. c, Detection of apoptotic cells by flow cytometric analysis. Cells were harvested from TNF−/− mice challenged with murine or human rTNF on day 14 after BLM. BAL was performed 6 h after airway challenge with TNF. Numbers represent percentage of annexin V- and/or propidium iodide-positive cells. Results are representative of five mice. d, Number of inflammatory cells in BALF harvested from TNF−/− mice challenged with various concentrations of murine rTNF on day 14 after BLM instillation. BAL was performed 48 h after airway challenge with murine TNF. Data represent mean ± SEM (n = 5 for each group). ∗∗, p < 0.05.
- FIGURE 8.
Effects of long-term challenge with murine rTNF on pulmonary inflammation. Challenge via airways with 250 ng/ml murine TNF solution was started on day 14 after BLM and repeated twice weekly for 4 wk. a and b, Histological analysis of lungs harvested from murine TNF-treated (a) or untreated (b) TNF−/− mice. The paraffin sections were H&E stained and photographed at ×40 magnification. Lungs were harvested on day 42 after BLM. Only unilateral lungs were macroscopically and histologically damaged in murine TNF-untreated TNF−/− mice. Results are representative of three mice in each group. c, Percentage of survival of mice. TNF−/− mice challenged with mouse TNF (○; n = 8) or sterile PBS (•; n = 8). TNF+/+ mice (Δ; n = 8) for control. It was confirmed that bilateral lungs were histologically damaged in all mice at the time of death or sacrifice on day 42 after BLM instillation. Data from two experiments were combined.
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