Duck lymphocytes—V. Transformation responses to phorbol ester and calcium ionophore

Abstract

1. Lymphocytes purified from duck blood and spleen were cultured in the presence of phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore A23187. Stimulation was assessed by the incorporation of [ 3H]thymidine after 3 days' culture. 1. 2. PMA stimulated over a wide range of concentrations, with maximum stimulation at final concentrations of 5 × 10 −7−5 × 10 −8 M/litre. A23187 was effective in the range 5 × 10 −6−5 × 10 −7 M/litre and also, in some experiments using spleen lymphocytes, at 5 × 10 −11−5 × 10 −12 M litre . 2. 3. Synergism was observed between PMA and A23187, the pattern depending on the concentrations of these reagents employed. Synergism was also observed between PMA and suboptimum concentrations of phytohaemagglutinin (PHA), wheat germ agglutinin (WGA), pokeweed mitogen (PWM) and Bandeiraea simplicifolia seed extract (BSS), but not with concanavalin A (Con A), lentil lectin (LL) or Helix pomatia lectin (HP). Similarly, synergism occurred between A23187 and WGA or PWM, but not with PHA, BSS, Con A, LL or HP. 3. 4. Mitomycin C and cycloheximide inhibited the response of duck lymphocytes to PMA, A23187 and lectins. Cyclosporin A inhibited responses to lectins but not to PMA or A23187. Neither hydrocortisone nor indomethacin inhibited responses to lectins, PMA or A23187. 4. 5. These results indicate that activation of duck lymphocytes occurs by virtue of similar intracellular messenger pathways to those operating in mammalian lymphocytes.