Optimum conditions for the turkey lymphocyte transformation test.
Optimum conditions for turkey lymphocyte transformation tests were determined. Thrice-washed turkey buffy-coat cells obtained after slow centrifugation (40 x g, 10 minutes) responded well to mitogenic stimulation. Turkey lymphocytes isolated on Ficoll-containing separation media largely lost their ability to respond to mitogens. Maximum responses were obtained with 2 x 10(7) lymphoid cells/ml. Responses to the mitogens were greatest when bovine fetal serum was used at a 2.5% concentration or pooled turkey serum and autologous plasma were used at a 1.25% concentration. Higher concentrations of turkey serum or plasma decreased the responses when sub-optimum doses of concanavalin-A (Con A) or phytohemagglutinin-P (PHA-P) were used. Serum-free cultures gave higher stimulation indices than cultures with serum only when sub-optimum doses of Con A or PHA-P were used. Optimum mitogen concentrations varied with individual birds, timing of the culture, temperature of incubation, and serum concentration in the cultures. Responses were usually greatest with final concentrations of 5 micrograms Con A/ml, 10 micrograms PHA-P/ml, and 20 micrograms pokeweed mitogen (PWM)/ml and when the cultures were incubated in 96-well microplates at 40 C in humidified air with 5% CO2 for 40-42 hours with pulsing with 3H-thymidine during the final 16 hours of incubation.
Barta
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Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Barta
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Domermuth
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Pierson
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FW
D000818:Animals; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D007106:Immune Sera / Q000276:immunology; D007958:Leukocyte Count / Q000662:veterinary; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D008934:Mitogens / Q000008:administration & dosage; D013696:Temperature; D014422:Turkeys / Q000097:blood* / Q000276:immunology
D007106:Immune Sera; D008934:Mitogens