Neutrophil elastase promotes macrophage cell adhesion and cytokine production through the integrin-Src kinases pathway

There are a number of respiratory diseases characterized by the presence of excess neutrophil elastase (NE) activity in tissues, including cystic fibrosis and chronic obstructive pulmonary disease (COPD). NE is considered a primary contributor to disease development, but the precise mechanism has yet to be fully determined. We hypothesized that NE alters the function of macrophages (M phi) which play a critical role in many physiological processes in healthy lungs. We demonstrate that monocyte-derived M phi exposed to NE releases active matrix metalloproteinases (MMPs), increase expression of pro-inflammatory cytokines TNF alpha, IL-1 beta, and IL-8, and reduce capacity to phagocytose bacteria. Changes in M phi function following NE treatment were accompanied by increased adhesion and cytoskeleton re-arrangement, indicating the possibility of integrin involvement. To support this observation, we demonstrate that NE induces phosphorylation of kinases from the Src kinase family, a hallmark of integrin signaling activation. Moreover, pretreatment of M phi with a specific Src kinase inhibitor, PP2 completely prevents NE-induced pro-inflammatory cytokine production. Taken together these findings indicate that NE participates in lung destruction not only through direct proteolytic degradation of matrix proteins, but also through activation of M phi inflammatory and proteolytic functions.