Supplementary Figure Legends
Figure S1. (A) Serum IgE levels in naïve and T. muris(Tm)-infected mice measured by ELISA. Flow cytometric analysis for mast cells (B) (CD45+Lineage (Lin)FcεRIα+IgE+CD49b+/-ckit+) in naïve and Tm-infected mice showing percentage of Lin- in (C) cecum and (D) spleen. (E) MLN basophil gating (CD45+Lineage (Lin)FcεRIα+IgE+CD49b+ckit-). Basophil percentage of (F) Lin- and (G) CD45+ MLN cells. Data from all combined experiments are presented as the mean ± SEM. Statistical analysis was performed using a linear fixed-effect model with pairwise comparison; P ≤ 0.05, **P ≤ 0.01, *** P ≤ 0.001. n=2-3 for each time point per experiment; 3 independent experiments.
Figure S2. Flow cytometric analysis of basophils in the spleen in naïve and T. muris (Tm)-infected mice, displaying (A) representative plots for Notch2 staining, (B) percent Notch2+ basophils, and (C) Notch2 geometric mean fluorescence intensity (gMFI) on spleen basophils. Data from all combined experiments are presented as the mean ± SEM. Statistical analysis was done using a linear fixed-effect model with pairwise comparison; ***P ≤ 0.001. n=2-3 for each time point per experiment; 3 independent experiments.
Figure S3. Real time PCR analysis of proximal colon homogenate from naïve and T. muris (Tm)-infected mice, quantifying (A)Il4 and (B) Il13 (normalized to Actb ). Data from all combined experiments are presented as the mean ± SEM. Statistical analysis was done using a linear fixed-effect model with pairwise comparison; *P ≤ 0.05. n=2-3 for each time point per experiment; 3 independent experiments.
References
1. Giacomin PR, Siracusa MC, Walsh KP, Grencis RK, Kubo M, Comeau MR, et al. Thymic stromal lymphopoietin-dependent basophils promote Th2 cytokine responses following intestinal helminth infection. J Immunol. 2012;189(9):4371-8.
2. Grencis RK. Immunity to helminths: resistance, regulation, and susceptibility to gastrointestinal nematodes. Annu Rev Immunol. 2015;33:201-25.
3. Harris NL, Loke P. Recent Advances in Type-2-Cell-Mediated Immunity: Insights from Helminth Infection. Immunity. 2018;48(2):396.
4. Maizels RM, Pearce EJ, Artis D, Yazdanbakhsh M, Wynn TA. Regulation of pathogenesis and immunity in helminth infections. J Exp Med. 2009;206(10):2059-66.
5. Perrigoue JG, Marshall FA, Artis D. On the hunt for helminths: innate immune cells in the recognition and response to helminth parasites. Cell Microbiol. 2008;10(9):1757-64.
6. von Moltke J, Locksley RM. I-L-C-2 it: type 2 immunity and group 2 innate lymphoid cells in homeostasis. Curr Opin Immunol. 2014;31:58-65.
7. Webb LM, Oyesola OO, Fruh SP, Kamynina E, Still KM, Patel RK, et al. The Notch signaling pathway promotes basophil responses during helminth-induced type 2 inflammation. J Exp Med. 2019;216(6):1268-79.
8. Zaph C, Cooper PJ, Harris NL. Mucosal immune responses following intestinal nematode infection. Parasite Immunol. 2014;36(9):439-52.
9. Antignano F, Mullaly SC, Burrows K, Zaph C. Trichuris muris infection: a model of type 2 immunity and inflammation in the gut. J Vis Exp. 2011(51).
10. Cliffe LJ, Grencis RK. The Trichuris muris system: a paradigm of resistance and susceptibility to intestinal nematode infection. Adv Parasitol. 2004;57:255-307.
11. Else KJ, Hultner L, Grencis RK. Cellular immune responses to the murine nematode parasite Trichuris muris. II. Differential induction of TH-cell subsets in resistant versus susceptible mice. Immunology. 1992;75(2):232-7.
12. Hurst RJ, Else KJ. Trichuris muris research revisited: a journey through time. Parasitology. 2013;140(11):1325-39.
13. Klementowicz JE, Travis MA, Grencis RK. Trichuris muris: a model of gastrointestinal parasite infection. Semin Immunopathol. 2012;34(6):815-28.
14. Else KJ, Keiser J, Holland CV, Grencis RK, Sattelle DB, Fujiwara RT, et al. Whipworm and roundworm infections. Nat Rev Dis Primers. 2020;6(1):44.
15. Artis D, Potten CS, Else KJ, Finkelman FD, Grencis RK. Trichuris muris: host intestinal epithelial cell hyperproliferation during chronic infection is regulated by interferon-gamma. Exp Parasitol. 1999;92(2):144-53.
16. Else K, Wakelin D. The effects of H-2 and non-H-2 genes on the expulsion of the nematode Trichuris muris from inbred and congenic mice. Parasitology. 1988;96 ( Pt 3):543-50.
17. Else KJ, Grencis RK. Cellular immune responses to the murine nematode parasite Trichuris muris. I. Differential cytokine production during acute or chronic infection. Immunology. 1991;72(4):508-13.
18. Qu SY, He YL, Zhang J, Wu CG. Transcription factor RBP-J-mediated signalling regulates basophil immunoregulatory function in mouse asthma model. Immunology. 2017;152(1):115-24.
19. Qu SY, Lin JJ, Zhang J, Song LQ, Yang XM, Wu CG. Notch signaling pathway regulates the growth and the expression of inflammatory cytokines in mouse basophils. Cell Immunol. 2017;318:29-34.
20. Sullivan BM, Locksley RM. Basophils: a nonredundant contributor to host immunity. Immunity. 2009;30(1):12-20.
21. Voehringer D. The role of basophils in helminth infection. Trends Parasitol. 2009;25(12):551-6.
22. Webb LM, Tait Wojno ED. Notch Signaling Orchestrates Helminth-Induced Type 2 Inflammation. Trends Immunol. 2019;40(6):538-52.
23. Eberle JU, Voehringer D. Role of basophils in protective immunity to parasitic infections. Semin Immunopathol. 2016;38(5):605-13.
24. Noti M, Wojno ED, Kim BS, Siracusa MC, Giacomin PR, Nair MG, et al. Thymic stromal lymphopoietin-elicited basophil responses promote eosinophilic esophagitis. Nat Med. 2013;19(8):1005-13.
25. Ohnmacht C, Schwartz C, Panzer M, Schiedewitz I, Naumann R, Voehringer D. Basophils orchestrate chronic allergic dermatitis and protective immunity against helminths. Immunity. 2010;33(3):364-74.
26. Siracusa MC, Saenz SA, Hill DA, Kim BS, Headley MB, Doering TA, et al. TSLP promotes interleukin-3-independent basophil haematopoiesis and type 2 inflammation. Nature. 2011;477(7363):229-33.
27. Sullivan BM, Liang HE, Bando JK, Wu D, Cheng LE, McKerrow JK, et al. Genetic analysis of basophil function in vivo. Nat Immunol. 2011;12(6):527-35.
28. Voehringer D, Reese TA, Huang X, Shinkai K, Locksley RM. Type 2 immunity is controlled by IL-4/IL-13 expression in hematopoietic non-eosinophil cells of the innate immune system. J Exp Med. 2006;203(6):1435-46.
29. Webb LM, Tait Wojno ED. The role of rare innate immune cells in Type 2 immune activation against parasitic helminths. Parasitology. 2017;144(10):1288-301.
30. Maillard I, He Y, Pear WS. From the yolk sac to the spleen: New roles for Notch in regulating hematopoiesis. Immunity. 2003;18(5):587-9.
31. Else KJ, Finkelman FD, Maliszewski CR, Grencis RK. Cytokine-mediated regulation of chronic intestinal helminth infection. J Exp Med. 1994;179(1):347-51.
32. Poorafshar M, Helmby H, Troye-Blomberg M, Hellman L. MMCP-8, the first lineage-specific differentiation marker for mouse basophils. Elevated numbers of potent IL-4-producing and MMCP-8-positive cells in spleens of malaria-infected mice. Eur J Immunol. 2000;30(9):2660-8.
33. Wada T, Ishiwata K, Koseki H, Ishikura T, Ugajin T, Ohnuma N, et al. Selective ablation of basophils in mice reveals their nonredundant role in acquired immunity against ticks. J Clin Invest. 2010;120(8):2867-75.
34. Siracusa MC, Comeau MR, Artis D. New insights into basophil biology: initiators, regulators, and effectors of type 2 inflammation. Ann N Y Acad Sci. 2011;1217:166-77.
35. Cruickshank SM, Deschoolmeester ML, Svensson M, Howell G, Bazakou A, Logunova L, et al. Rapid dendritic cell mobilization to the large intestinal epithelium is associated with resistance to Trichuris muris infection. J Immunol. 2009;182(5):3055-62.
36. Sahputra R, Ruckerl D, Couper KN, Muller W, Else KJ. The Essential Role Played by B Cells in Supporting Protective Immunity Against Trichuris muris Infection Is by Controlling the Th1/Th2 Balance in the Mesenteric Lymph Nodes and Depends on Host Genetic Background. Front Immunol. 2019;10:2842.
37. Taylor BC, Zaph C, Troy AE, Du Y, Guild KJ, Comeau MR, et al. TSLP regulates intestinal immunity and inflammation in mouse models of helminth infection and colitis. J Exp Med. 2009;206(3):655-67.
38. Chenery AL, Antignano F, Hughes MR, Burrows K, McNagny KM, Zaph C. Chronic Trichuris muris infection alters hematopoiesis and causes IFN-gamma-expressing T-cell accumulation in the mouse bone marrow. Eur J Immunol. 2016;46(11):2587-96.
39. Hasnain SZ, Evans CM, Roy M, Gallagher AL, Kindrachuk KN, Barron L, et al. Muc5ac: a critical component mediating the rejection of enteric nematodes. J Exp Med. 2011;208(5):893-900.