Optimization and kinetics of in vitro stimulation of carp (Cyprinus carpio L.) leukocytes

The optimization of a leukocyte stimulation microassay with carp (Cyprinus carpio L.) leukocytes is described. Leukocytes were isolated from the thymus, anterior kidney, spleen, mid-kidney and peripheral blood. Leukocyte cultures were stimulated with PHA-P, LPS (Escherichia coli 055: B5) PWM, ConA and PPD from Mycobacterium fortuitum. The optimum incubation temperature for leukocyte cultures differed 3.5 days for leukocyte cultures derived from lymphoid organs and 4.5 days for peripheral blood lymphocyte cultures. Leukocytes from various organ sources showed similar reactivity patterns to stimulation in vitro by different mitogens. The results of these mitogen stimulations did not present sufficient arguments in favour of compartmentation.