Optimization of an in vitro assay which measures the proliferation of duck T lymphocytes from peripheral blood in response to stimulation with PHA and ConA

Abstract

The in vitro proliferative responses of duck PBMCs purified from Ficoll-Paque density gradients to the mitogens PHA and ConA show a great deal of duck-to-duck variation. Better responses were consistently obtained by using nylon wool fractionation to increase the proportion of duck T lymphocytes in PBMC preparations and then culturing these preparations with homologous monocytes, purified from PBMC preparations by their adherence properties. We have also established that the addition of homologous red blood cells enhances the in vitro proliferative responses of duck T lymphocytes, especially when limiting doses of PHA and ConA are used. Duck T lymphocytes showed greater and more consistent proliferation when cultures were incubated at 37 degrees C as compared to incubation at 41.6 degrees C. The improved consistency of higher proliferative responses with this assay should make it more suitable for detecting in vitro proliferative responses of antigen-specific T lymphocytes, as a measure of in vivo induced cell mediated immune responses.