Isolation, characterization and in vitro mitogenic stimulation of peripheral blood lymphocytes from an American buffalo.
A rapid and reproducible method is described for the isolation and characterization of leukocytes from the peripheral blood of an American buffalo (bison). Centrifugation of the buffy coat cells on a Percoll gradient (1.079 g/mL) at 650 x g for 20 min resulted in the separation and high yields of pure viable leukocytes. The sheep erythrocyte-rosetting technique (ER) showed that 59% of the cells were ER+ (T lymphocytes). Fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin and FITC-conjugated concanavalin A revealed 77% and 89% positive cells, respectively. The isolated leukocytes contained adherent accessory cells and functionally active T and B lymphocytes which proliferated in response to both T and B cell mitogens and to exogenous recombinant bovine interleukin-2 in the absence and/or presence of the thiol compound 2-mercaptoethanol.
Nagi
A M
AM
Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon.
Babiuk
L A
LA
D000818:Animals; D001188:Artiodactyla / Q000097:blood*; D002469:Cell Separation; D002470:Cell Survival; D005260:Female; D007958:Leukocyte Count / Q000662:veterinary; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D010641:Phenotype