Bovine platelet adhesion is enhanced by leukotoxin and sialoglycoprotease isolated from Pasteurella haemolytica A1 cultures.
Platelet and fibrin deposits are among characteristic changes observed in lung alveoli of cattle with pasteurellosis induced by Pasteurella haemolytica (biotype A, serotype 1). To determine whether the platelet function could be directly affected by protein products produced by the bacterium, the effects of leukotoxin and O-sialoglycoprotease, culture supernatant antigen secreted by Pasteurella haemolytica A1, on bovine platelet activation were examined by evaluating the enhancement of platelet adhesion to a negatively charged surface relative to untreated control samples. The glycoprotease, or the leukotoxin, was added to plasma free suspensions of bovine platelets and platelet adhesion assessed by two parameters: (i) the number of 3H-adenine-labeled adherent platelets and (ii) the morphology of unlabeled platelets adhering to the charged surface under scanning electron microscopy (SEM). In the presence of calcium, the glycoprotease produced a dose-dependent increase in adhesion. At a concentration of 4.0 micrograms glycoprotease extract protein per 10(7) platelets, a 2-fold increase in adhesion was observed which was similar to the increase in adhesion induced by 0.10 units of thrombin, a known platelet agonist. Both increased platelet adhesion and platelet aggregation were observed with 0.8 microgram glycoprotease extract protein in the presence of calcium. The response of the bovine platelet suspensions to leukotoxin extract protein was dependent on the dosage of the leukotoxin. Adhesion was enhanced at dosages of 25 micrograms leukotoxin protein per 10(7) platelets and below, while at dosages of 50 micrograms and above adhesion was suppressed. Thus, the two proteins secreted by P. haemolytica may interact directly with bovine platelets to initiate platelet aggregation and fibrin formation in alveolar tissue in pneumonic pasteurellosis.
Nyarko
K A
KA
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Canada.
Coomber
B L
BL
Mellors
A
A
Gentry
P A
PA
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D002417:Cattle; D004357:Drug Synergism; D005098:Exotoxins / Q000302:isolation & purification / Q000494:pharmacology*; D005260:Female; D066298:In Vitro Techniques; D016978:Mannheimia haemolytica / Q000201:enzymology / Q000254:growth & development*; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D008855:Microscopy, Electron, Scanning; D010973:Platelet Adhesiveness / Q000187:drug effects*; D013917:Thrombin / Q000494:pharmacology
D001427:Bacterial Toxins; D005098:Exotoxins; C018579:leukotoxin; D013917:Thrombin; D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase