Optimal conditions for measurement of blastogenic responses of chickens to concanavalin A in whole blood assays.
One-day-old chicks (Cobb 500) were raised in wire cages in clean isolated conditions and fed on a non-medicated diet. Heparinised peripheral blood samples were taken from the cutanea ulnaris (wing) vein. The effect of various parameters including concentration of concanavalin A (Con A), dilution of blood in medium, volume of blood dilution per culture, type of media, percentage of serum, incubation period, concentration of thymidine, pulse-labelling period, and age of chickens on the blastogenic response of chickens to Con A was evaluated. The results showed that despite a certain degree of individual variation among responses, the highest response was to 0.4 microgram Con A per culture at a blood dilution of 1/50 in serum-free RPMI 1640 during incubation for 90 h at 40 degrees C in a humidified atmosphere containing 5% CO2. Cultures were pulsed with 0.2 microCi thymidine per culture for 18 h prior to harvesting. The stimulation response of chicken whole blood lymphocytes of chickens to Con A was prominent from 3 weeks of age onwards and the response measured in chickens 3 weeks of age was slightly higher than that of subsequent weeks. The possible applications of this technique in studying the pathogenesis of avian diseases and for quantitative assessment of circulating T cells of chickens are discussed.
Talebi
A
A
Department of Veterinary Microbiology and Parasitology, Faculty of Veterinary Medicine, University College Dublin, Ireland.
Torgerson
P R
PR
Mulcahy
G
G
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens / Q000097:blood* / Q000276:immunology; D003208:Concanavalin A / Q000494:pharmacology*; D004261:DNA Replication; D004305:Dose-Response Relationship, Drug; D007111:Immunity, Cellular; D037102:Lectins / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects*; D013601:T-Lymphocytes / Q000187:drug effects*; D013936:Thymidine
D037102:Lectins; D003208:Concanavalin A; D013936:Thymidine