A232 TUFT CELLS COORDINATE RAPID EXPULSION OF THE TAPEWORM H. DIMINUTA BUT ARE NOT REQUIRED FOR ENHANCED IMMUNITY AGAINST THE NEMATODE, H. POLYGYRUS, IN MICE PREVIOUSLY INFECTED WITH H. DIMINUTA

Abstract Background The tuft cell is an important sentinel that monitors the gut lumen and coordinates immunity against parasitic nematodes. We showed small intestinal tuft cell hyperplasia 11 days post-infection (dpi.) with the tapeworm Hymenolepis diminuta: a time when the parasite is no longer present in murine hosts. This may be a way by which the host protects itself from subsequent helminth-infections, a common phenomenon in parasite-endemic world regions. We test this supposition using Pou2f3-/- mice that lack tuft cells. Aims To test the hypothesis that tuft cells are important in the anti-worm response in H. diminuta ( H.d.)-infected mice subsequently infected with the nematode parasite Heligosomoides polygyrus ( H.p.). Methods Male C57BL6 and Pou2f3-/- mice (8–12 weeks) were infected with 5 H.d. cysticercoids ± 200 H.p. larvae at 10 dpi with H. diminuta (non H.p. mice - control). Upon necropsy at 24 dpi H. diminuta (i.e. 14 dpi H.p. in co-infected mice), both worms were ennumerated in small intestinal washings, H.p. granulomas examined and fecal egg counts performed. Small intestinal segments were stained for tuft (DCLK1+) and goblet cells (PAS+). As a surrogate of successful infection, IL-4 and IL-10 were measured in supernatants from concanavalin-A treated splenocytes. Results Wild-type (WT) mice expel H. diminuta by 11 dpi and this was delayed in Pou2f3-/-mice, with worms readily detectable at 14 dpi and absent by 21 dpi. Despite the delayed expulsion, both WT and Pou2f3-/- mice showed increased splenic production of IL-4 and IL-10; however, unlike WT mice, H. diminuta-infected Pou2f3-/- mice show no increase in jejunal goblet cell numbers. Mice infected with H. diminuta displayed a degree of increased resistance to H.p.-infection defined by reduced worm and egg burdens, and increased granuloma formation in comparison to H.p.-only infected animals. In this sequential co-infection model, there were no significant differences between WT and Pou2f3-/- mice in the response to H.p. Conclusions The absence of tuft cells slows expulsion of H. diminuta from its non-permissive mouse host and correlates with diminished goblet cell hyperplasia. Hypothesizing that H. diminuta-evoked tuft cell hyperplasia would enhance the immune response to a subsequent infection with an unrelated nematode parasite proved incorrect. While H. diminuta-infected mice were partially protected from H.p., response was similar in WT and Pou2f3-/- mice. Thus tuft cells are important in worm detection: yet, our co-infection data suggests that other events initiated by the primary worm infection impact the outcome of subsequent infection with a different helminth and tuft cells have a limited, if any, role to play in this helminth-host-helminth interaction. Funding Agencies CIHRNSERC, Eye’s High International doctoral scholarship