Effect of leukotoxin of Mannheimia haemolytica and LPS of E. coli on secretory response of bovine neutrophils in vitro.
To evaluate the role of leukotoxin (LKT) of Mannheimia haemolytica and lipopolysaccharide (LPS) of E. coli 055:B5 in pathogenesis of bovine respiratory disease (BRD) we investigated their in vitro effects on cultured bovine neutrophils. Functional parameters of neutrophils including degranulation, generation of superoxide, and nitric oxide were distorted in response to both toxins. The most essential reaction of neutrophils was found in respect to release of elastase after addition of LKT as well as LPS at concentration of 300 microg/ml. Moreover, we observed an increased release of myeloperoxidase (MPO) and alkaline phosphatase (ALK-P) from polymorphonuclear cells (PMN) after addition of LKT and LPS. We also found enhanced superoxide generation by bovine neutrophils after exposure to different concentrations of LKT and LPS. In cultures of PMN treated with LKT, concentration of nitrite increased with growing concentrations of LKT. Lower values of nitrite were obtained in cultures exposed to LPS. Partial lysis of PMN, determined by LDH (lactate dehydrogenase) leakage, started at concentration of 300 microg/ml for both toxins, meanwhile LKT concentration above 300 microg/ml was lethal. Our study has revealed that neutrophils in response to both toxins exaggerate release of analysed substances, which participate in worsening the course of the disease and play a role in lung injury during BRD. Toxins introduced to the cultural medium stimulate release of studied constituents from neutrophils by combined activation and lysis of neutrophils.
Wessely-Szponder
J
J
Department of Pathophysiology Faculty of Veterinary Medicine, University of Agriculture, Akademicka 12, 20-033 Lublin, Poland. joanna.wessely@ar.lublin.pl
Urban-Chmiel
R
R
Wernicki
A
A
Bobowiec
R
R
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001427:Bacterial Toxins / Q000633:toxicity*; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D004926:Escherichia coli / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary; D005098:Exotoxins / Q000633:toxicity; D008070:Lipopolysaccharides / Q000633:toxicity*; D016978:Mannheimia haemolytica / Q000472:pathogenicity*; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D012766:Pasteurellosis, Pneumonic / Q000382:microbiology; D009195:Peroxidase / Q000378:metabolism; D012141:Respiratory Tract Infections / Q000382:microbiology / Q000662:veterinary*
D001427:Bacterial Toxins; D005098:Exotoxins; D008070:Lipopolysaccharides; C018579:leukotoxin; D009195:Peroxidase; D000469:Alkaline Phosphatase