Functional analyses of lymphocytes and granulocytes isolated from the thymus, spiral valve intestine, spleen, and kidney of juvenile Australian lungfish, Neoceratodus forsteri

Our current understanding of the lungfish immune system is limited. This study is characterizing the immune cells separated from primary and secondary immune organs of the Australian lungfish, Neoceratodus forsteri. Our functional studies utilized flow cytometry to study the immune cells extracted from the thymus, spiral valve intestine, spleen, and kidney. The different characteristics of lymphocytes and granulocytes were analyzed by utilization of viability, phagocytosis, oxidative burst, and apoptosis assays. Most of the nonviable intestinal cells were lymphocytes. Depending on the organ, 6-25% of the total population, predominantly granulocytes, underwent phagocytosis where the splenic cells were the most and intestinal cells the least phagocytic cells. Cells responded positively but differently to stimulation with phorbol myristate acetate (PMA) to produce radical oxygen species, an indication of their oxidative burst activity, which was mainly associated with granulocytes. Although cells were induced by dexamethasone to undergo apoptosis, such an induction did not follow a consistent pattern of dose of dexamethasone or incubation time between the different organs. In the absence of monoclonal antibodies against lungfish immune cells, these functional flow cytometric analyses aid our understanding on the functionality of immune cells.