OPTIMIZATION OF AN IN-VITRO ASSAY WHICH MEASURES THE PROLIFERATION OFDUCK T-LYMPHOCYTES FROM PERIPHERAL-BLOOD IN RESPONSE TO STIMULATION WITH PHA AND CONA

The in vitro proliferative responses of duck PBMCs purified from Ficol l-Paque density gradients to the mitogens PHA and ConA show a great de al of duck-to-duck variation, Better responses were consistently obtai ned by using nylon wool fractionation to increase the proportion of du ck T lymphocytes in PBMC preparations and then culturing these prepara tions with homologous monocytes, purified from PBMC preparations by th eir adherence properties. We have also established that the addition o f homologous red blood cells enhances the in vitro proliferative respo nses of duck T lymphocytes, especially when limiting doses of PHA and ConA are used, Duck T lymphocytes showed greater and more consistent p roliferation when cultures were incubated at 37 degrees C as compared to incubation at 41.6 degrees C, The improved consistency of higher pr oliferative responses with this assay should make it more suitable for detecting in vitro proliferative responses of antigen-specific T lymp hocytes, as a measure of in vivo induced cell mediated immune response s. (C) 1997 Elsevier Science Ltd.