Stimulated rabbit alveolar macrophages secrete a growth factor for type II pneumocytes

The type II pneumocyte plays a principle role in the maintenance and repair of the pulmonary alveolar epithelium by increasing its rate of proliferation under conditions of epithelial damage. This investigation examined the role of the alveolar macrophage in the control of type II cell division through its ability to produce specific growth factors when activated in vitro. Type II cells were isolated from adult male rabbits and cultured in the presence of media and matrix that support cell proliferation. Proliferation was assessed by cell counting and pulsing with [3H]thymidine, followed by measurements of labeling index and TCA-insoluble radioactivity. Alveolar macrophages were cultured in serum-free media in the presence of a particulate stimulus. Conditioned media was diluted and added to type II cell cultures. Conditioned media from stimulated macrophage cultures was found to double basal type II cell proliferation, whereas media from unstimulated macrophage cultures had no effect. Macrophage production of type II cell growth-promoting activity was dependent on the concentration of the stimulus and the length of the incubation. Investigation into the identity of the growth-regulating protein established that it is heat labile, insensitive to reduction and acidic conditions, and sensitive to trypsin digestion. Its molecular weight appears to be greater than or equal to 25 kD. Addition of several characterized growth factors to type II cell cultures demonstrated that other known growth-promoting products of macrophages do not act as type II cell growth factors. The evidence presented suggests that in vitro activated alveolar macrophages produce a type II cell growth factor that may play a critical role in mediating repair of the alveolar epithelium.