Anterior kidney and splenic cells were taken from rainbow trout and splenic cells from BALB/c mice immunized with a T-dependent (sheep red blood cells) or T-independent (DNP-Ficoll) antigen. The cells were incubated at different temperatures in Jerne plaque assays (direct or passive haemolytic plaque assays). The optimum numbers of in vitro plaque-forming cells (PFC) after incubation with homologous complement were directly correlated with normal body temperatures of the respective species. The optimum incubation temperature was 37°C for mouse cells and 10°C for fish cells. Incubation of mouse cells at lower temperatures of 30, 20, 10, 4 or 0°C appeared to yield a direct line reduction in numbers of PFC. Trout cells developed significantly fewer PFC at 4 and 20°C and none at 30°C or above; however, significant numbers still appeared at 0°C. More PFC per million white blood cells were obtained from the anterior kidney; however, related to temperatures, no differences in development of numbers of PFC could be seen between the spleen and anterior kidney cells of trout. When the incubation time was lengthened for both trout and mouse cells held at low temperatures, the numbers of PFC approached those of the cells incubated at the optimum temperatures for 10 h.
