1. 1 ZIB Paper presentation 07.02.2011 Wiebke Ginter Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity Daniel R. Neill1*, See Heng Wong1*, Agustin Bellosi1*, Robin J. Flynn1, Maria Daly1, Theresa K. A. Langford1, Christine Bucks2, Colleen M. Kane2, Padraic G. Fallon3, Richard Pannell1, Helen E. Jolin1 & Andrew N. J. McKenzie1

2. 2  Plays role in helminth infection and asthma  Eosinophilia, mucus secretion, IgE production  IL-4, IL5, IL-13  Produced by basophils, eosinophils and mast cells  Role in Nippostrongylus brasiliensis expulsion unknown Nippostrongylus brasiliensis intestinal nematode of rodents closely related to the human hookworms (Ancylostoma and Necator) infect 740 million people in tropical countries N. brasiliensis widely used model for nematode parasites Maizels, 2003, Nat. Rev. Immunol Type II immunity and Question

3. 3 IL-13 is critical in immune response to helminths and asthma Which cell types to produce this cytokine? Supp. 1a  Plays role in helminth infection and asthma  Eosinophilia, mucus secretion, IgE production  IL-4, IL5, IL-13  Produced by basophils, eosinophils and mast cells  Role in Nippostrongylus brasiliensis expulsion unknown

4. 4 CD4 + splenocytes Supp.1 Th1 conditionsTh2 conditions

5. 5 Localisation of eGFP + cells Fig. 1c Supp. 2b

6. 6 no T cell no APC no gd T cell no NK/NKT cell no DC mono… no macroph, granolo no eos no mast no pDC no B cell Surface markers Supp. 2a,c

7. 7 New cells are no mast, B, T, NK or LTi cells

8. 8 Induction of Nuocytes by IL-25 and IL-33 Fig. 1 IL-25 receptorSU of IL-33 receptor  Direct action of these IL  Redundant action Nuocytes: Lin - eGFP + T1/ST2 + IL17BR +

9. 9 Presence in imunosurveillance sites Supp. 4

10. 10 Helminth infection Small intestine Supp. 2d Fig. 2a

11. 11 IL25 and IL33: partially redundant roles Il17br -/- Il1rl1 -/- IL17br -/- Il1rl1 -/- No IL25RNo SU IL33RNo IL25R, IL33R Fig.2

12. 12 Nuocyte Cultures Fig. 3  IL33 and IL7 needed for expansion  Most cell surface markers maintained  Did not differentiate Supp. 8 Nuocytes: Lin - ICOS + T1/ST2 + IL17BR + IL-7Rα +

13. 13 Cytokine profile Fig. 3 Supp. 9

14. 14 Adoptive transfer of cultured nuocytes into Il17br-/- mice restores an IL25- responsive phenotype Il17br-/-

15. 15 Restoring protective IR in IL17BR -/- mice Fig. 4

16. 16 IL13 of nuocytes is necessary and sufficient for worm expulsion Fig. 4 IL13 from nuocytes is the only important source

17. 17 Dialogue between T cells and nuocytes Fig. 4 T cellsNuocytes Enhance cytokines, augment resp Maintain: expansion survival

18. 18 Summary new class of cells: Nuocytes important for the type-2 immunity: protection from parasitic helminths release of type II cytokines, activating eosinophiles and goblet cells and secreting mucus and IgE Key source of IL-13 release during early steps of helminth infection Expand rapidly in response to IL-25 and IL-33 (type-2 initiators) Lack: nuocytes expansion ↓, defect in worm expulsion present in immunosurveillance sites Secrete high levels of IL13 and IL5 Enhance T cell responses highly specialized type type-2 regulatory cells New possible target for immune modulation? Asthma treatment? T cellsNuocytes Enhance cytokines, augment resp Maintain: expansion survival

19. 19 Supplementary Saenz, 2010, Trends in Immunology

20. 20 no T cellno APC no gd T cell no NK/NKT cell no DC mono… no macroph, granolo no eos no mast no pDC no B cell Surface markers Supp. 2a,c

21. 21 DR Neill et al. Nature 000, 1-4 (2010) doi:10.1038/nature08900 Adoptive transfer of wild-type nuocytes, but not IL13-deficient nuocytes, restores rapid worm expulsion in N. brasiliensis-infected Il17br _ / _ mice.