PMN-mediated immune reactions against Eimeria bovis

For successful in vivo infection, Eimeria bovis sporozoites have to traverse the mucosal layer of the ileum to infect lymphatic endothelial cells and may, thereby, be exposed to the interstitial fluid and to the lymph representing potential targets for leukocytes. To mimic this situation in vitro, we exposed E. bovis sporozoites to bovine PMN and found enhanced elimination of the parasites. Addition of immune serum clearly increased these reactions, whereas neonatal calf serum had no effect, thus proposing a PMN-derived antibody-dependent cytotoxicity. Scanning and transmission electron microscopy showed PMN engulfing sporozoites or extending filopodia towards them and occasionally incorporating the parasites. PMN reacted with enhanced transcription of IL-6, MCP-1, GROalpha, TNF-alpha, and iNOS genes after exposure to sporozoites while stimulation with merozoite-antigen, in addition, upregulated IL-8, IP-10 and IL-12 gene transcription. Furthermore, enhanced in vitro oxidative burst and phagocytic activities were observed after contact of PMN with viable sporozoites. To verify the potential role of PMN in the in vivo situation, we analysed the general phagocytic and oxidative burst activities of PMN obtained ex vivo from E. bovis experimentally infected calves. Enhanced levels of both activities were found early p.i. (1-5 days) and towards the end of the first schizogony (days 13-22 p.i.) underlining the in vitro data. Our results suggest that PMN-mediated, innate immune reactions play an important role in the early immune response to E. bovis infections in calves.