3-nitrotyrosine and Swine-Diseases

Cyclooxygenase-2 (COX-2) and nitric oxide synthase 2 (NOS2) were detected and localized in 20 pigs with ulcerative colitis caused by natural infection with Salmonella typhimurium. Evidence of NOS2 activity was determined by the formation of nitrotyrosine, a reaction product of peroxynitrite, in NOS2-expressing ulcerative colons by immunohistochemistry. Transcript RNA of COX-2 and NOS2 was consistently detected in colonic tissues from the 20 pigs with ulcerative colitis by using reverse transcription-polymerase chain reaction. Immunohistochemical signals for COX-2 and NOS2 were detected in the ulcerated area of all 20 pigs. Expression of COX-2 and NOS2 was identified continuously within inflammatory intestinal lesions but was minimal in unaffected regions of the colon of S. typhimurium-infected pigs. The immunohistochemistry of serial sections of intestine indicated that the majority of colons containing numerous COX-2-positive cells also had numerous NOS2-positive cells. Localization of NOS2 and a nitrotyrosine antigen was prominent in neutrophils and macrophages in the periphery of the lesions. Simultaneous detection of COX-2 and NOS2 RNA and protein indicated functional activity of prostaglandin and NO production in vivo. This study suggested that COX-2 and NOS2 expression may play a role in the pathophysiologic processes in ulcerative colitis caused by S. typhimurium.

Topics: Animals; Colitis, Ulcerative; Cyclooxygenase 2; DNA Primers; Immunohistochemistry; Isoenzymes; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Peroxynitrous Acid; Polymerase Chain Reaction; Prostaglandin-Endoperoxide Synthases; Prostaglandins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Salmonella Infections, Animal; Salmonella typhimurium; Swine; Swine Diseases; Tyrosine

Evidence of nitric oxide synthase (NOS) 2 activity was determined by formation of nitrotyrosine (a reaction product of peroxynitrite) and by activation of poly(ADP-ribose) synthetase (PARS) in NOS2-expressed pleuropneumonic lungs from 20 pigs naturally infected with Actinobacillus pleuropneumoniae using immunohistochemistry. Intense immunostaining for nitrotyrosine residue was seen within the lung lesions from A. pleuropneumoniae-infected pigs, but it was minimal in the unaffected parts of the lung from A. pleuropneumoniae-infected pigs and in the normal lung from control pigs. Staining was especially strong in neutrophils and macrophages in the periphery of the lesions and within the alveolar spaces. There was close cell-to-cell correlation when serial sections were examined by immunohistochemistry for NOS2 and nitrotyrosine in each of the 20 lung samples. Expression of PARS was always present within inflammatory lesions but was minimal in the unaffected lung of A. pleuropneumoniae-infected pigs. Macrophages in alveolar spaces frequently exhibited strong staining for PARS. Colocalization of nitrotyrosine and PARS antigen was especially prominent in macrophages in the periphery of lesions. NOS2 expression in pleuropneumonic areas associated with protein nitrosation and PARS suggests that NOS2 is functionally active during infections caused by A. pleuropneumoniae.

Topics: Actinobacillus Infections; Actinobacillus pleuropneumoniae; Animals; Antigens, Viral; Enzyme Induction; Immunohistochemistry; Lung; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Pleuropneumonia; Poly(ADP-ribose) Polymerases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Swine; Swine Diseases; Tyrosine

The expression of inducible nitric oxide synthase (iNOS), an enzyme that produces nitric oxide, was examined in the hearts of pigs infected with porcine enterovirus serotype 3 (PEV3). Piglets orally infected with PEV3 developed tremors and paralysis 3-7 days post-infection. Affected animals had pericarditis and myocarditis. There were fibrin and inflammatory cell infiltrates (macrophages and neutrophils) in the pericardial sac and myocardium. Immunohistochemically, the majority of inflammatory cells in the pericardial sac were positive for iNOS and nitrotyrosine, an end product of nitric oxide. These results suggest that iNOS is upregulated in the pericardial lesion, and that increased nitric oxide production plays an important role in the development of PEV3-induced pericarditis and myocarditis.

Topics: Animals; Animals, Newborn; Enterovirus Infections; Enteroviruses, Porcine; Immunohistochemistry; Microscopy, Electron; Myocardium; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Pericarditis; Pericardium; Swine; Swine Diseases; Tyrosine