Myeloid cell dynamics in bleomycin-induced pulmonary injury in mice; effects of anti-TNFα antibody

Bleomycin is a cancer therapeutic known to cause lung injury which progresses to fibrosis. Evidence suggests that macrophages contribute to this pathological response. Tumor necrosis factor (TNF)alpha is a macrophage-derived pro-inflammatory cytokine implicated in lung injury. Herein, we investigated the role of TNF alpha in macrophage responses to bleomycin. Treatment of mice with bleomycin (3 U/kg, i.t.) caused histopathological changes in the lung within 3 d which culminated in fibrosis at 21 d. This was accompanied by an early (3-7 d) influx of CD11b(+) and iNOS(+) macrophages into the lung, and Arg-1(+) macrophages at 21 d. At this time, epithelial cell dysfunction, defined by increases in total phospholipids and SP-B was evident. Treatment of mice with anti-TNF alpha antibody (7.5 mg/kg, i.v.) beginning 15-30 min after bleomycin, and every 5 d thereafter reduced the number and size of fibrotic foci and restored epithelial cell function. Flow cytometric analysis of F4/80(+) alveolar macrophages (AM) isolated by bronchoalveolar lavage and interstitial macrophages (IM) by tissue digestion identified resident (CD11b(-)CD11c(+)) and immature infiltrating (CD11b(+)CD11c(-)) AM, and mature (CD11b(+)CD11c(+)) and immature (CD11b(+)CD11c(-)) IM subsets in bleomycin treated mice. Greater numbers of mature (CD11c(+)) infiltrating (CD11b(+)) AM expressing the anti-inflammatory marker, mannose receptor (CD206) were observed at 21 d when compared to 7 d post bleomycin. Mature proinflammatory (Ly6C(+)) IM were greater at 7 d relative to 21 d. These cells transitioned into mature anti-inflammatory/pro-fibrotic (CD206(+)) IM between 7 and 21 d. Anti-TNF alpha antibody heightened the number of CD11b(+) AM in the lung without altering their activation state. Conversely, it reduced the abundance of mature proinflammatory (Ly6C(+)) IM in the tissue at 7 d and immature pro-fibrotic IM at 21 d. Taken together, these data suggest that TNF alpha inhibition has beneficial effects in bleomycin induced injury, restoring epithelial function and reducing numbers of profibrotic IM and the extent of pulmonary fibrosis.

Automated summary

The involvement of macrophages and TNF alpha in the pathological response to bleomycin-induced lung injury is crucial, and anti-TNF alpha antibody treatment can reduce fibrosis and restore epithelial cell function.