FLOW CYTOMETRIC DIFFERENTIATION OF AVIAN LEUKOCYTES AND ANALYSIS OF THEIR INTRACELLULAR CYTOKINE EXPRESSION
Résumé
A flow cytometric method for the identification of chicken blood leukocyte subpopulations and thrombocytes was developed. An anti-chicken CD45 phycoerythrin-labelled antibody was used to separate leukocytes from red blood cell nuclei. Leukocytes and thrombocytes were identified using a combination of their CD45-positivity and their typical side scatter properties. The identity of the CD45-positive cells was confirmed by sorting the subpopulations and subsequent light microscopic evaluation. In these differentiated cell populations, intracellular expression analysis of the pro-inflammatory cytokines interleukin-1β and IL-6 was subsequently optimised on whole blood after in vitro stimulation with lipopolysaccharide from Escherichia coli strain O127:B8.
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