Spontaneous hemolytic activity of rat alveolar lining material

During assays of the complement hemolytic activity in lavage fluids (LF) from humans and various laboratory animals (hamsters, rabbits, rats, guinea pigs), we have observed that rat bronchoalveolar lavages had a spontaneous, complement-independent, hemolytic activity to sheep red blood cells (SRBC). Rat lavage fluids were able to lyse sheep and autologous red blood cells at 2°C as well as at 37°C. Together with these observations, the inverse relationship that existed between the LF hemolytic activity and the calcium concentration in the incubation medium suggested that lysis could be due to the presence of large amounts of lysophospholipids in rat lavage fluid. However, thin layer chromatography did not reveal any abnormal amount in lysoderivative, whereas the free fatty acid (FFA) content was very high. Pure palmitic acid, at a concentration similar to that observed in LF from rat, was able to lyse SRBC (8.5µg lyzed 50% of 108 SRBC); its lytic activity decreased when Ca++ or bovine serum albumin was added to the incubation mixture. FFA through their detergent effect, appear to account for the hemolytic activity of the rat alveolar lining material.