Expression of nitric oxide synthase 2 and tumor necrosis factor alpha in swine naturally infected with Actinobacillus pleuropneumoniae

Nitric oxide synthase 2 (NOS2) and tumor necrosis factor alpha (TNF-alpha) were detected and localized in 15 pigs with naturally occurring pleuropneumonia by use of in situ hybridization with a nonradioactive digoxigenin-labeled cDNA probe. Two cDNA probes 491 and 219 base pairs for NOS2 and TNF-alpha, respectively, were generated by reverse transcription polymerase chain reaction. All 15 pigs infected with Actinobacillus pleuropneumoniae had distinct positive hybridization signals for NOS2 and TNF-alpha. Strong hybridization signals for both NOS2 and TNF-alpha were evident in degenerate alveolar leukocytes bordering zones of coagulative necrosis and in alveolar spaces. NOS2 nucleic acids were detected in neutrophils and macrophages. In situ hybridization of serial sections of lung tissue revealed numerous cells positive for NOS2 and TNF-alpha, suggesting that NOS2 and TNF-alpha expression may play a role in the pathophysiology of pleuropneumonia.