Chemotaxis of radiolabeled equine neutrophils.
A method for the isolation of equine neutrophils was developed using metrizamide cushions. A purity of greater than 95% was routinely obtained with greater than 90% viability. These cells were radiolabeled and tested for their chemotactic response in Boyden chambers to zymosan-activated equine serum, the partially purified equine complement component C5a, and formyl-L-methionyl-L-leucyl-L-phenylalanine. The time and ionic requirements for chemotaxis of radiolabeled equine neutrophils were investigated and maximal movement was observed at 2 hours' incubation and 1.0 mM Ca and 0.5 mM Mg. Dinitrophenol in a concentration range of 10(-2) to 10(-6) M also inhibited cell movement. A Zigmond-Hirsch checkerboard assay demonstrated the pronounced chemotactic activity of zymosan-activated equine serum and the partially purified equine C5a. The equine neutrophil did not respond to formyl-L-methionyl-L-leucyl-L-phenylalanine.
D000818:Animals; D002118:Calcium / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D002630:Chemotactic Factors / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D003182:Complement C5 / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D006736:Horses / Q000097:blood*; D006801:Humans; D066298:In Vitro Techniques; D008274:Magnesium / Q000502:physiology; D008793:Metrizamide / Q000494:pharmacology; D009239:N-Formylmethionine / Q000031:analogs & derivatives / Q000502:physiology; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009504:Neutrophils / Q000502:physiology*; D009842:Oligopeptides / Q000502:physiology; D011817:Rabbits / Q000097:blood; D013552:Swine / Q000097:blood; D015054:Zymosan / Q000494:pharmacology
D002630:Chemotactic Factors; D003182:Complement C5; D009842:Oligopeptides; D009239:N-Formylmethionine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015054:Zymosan; D008274:Magnesium; D008793:Metrizamide; D002118:Calcium