Eosinophils are specialized myeloid cells associated with allergy and helminth infections. Blood eosinophils demonstrate circadian cycling, as described over 80 years ago1, and are abundant in the healthy gastrointestinal tract. Although a cytokine, interleukin (IL)-5, and chemokines such as eotaxins mediate eosinophil development and survival2, and tissue recruitment3, respectively, the processes underlying the basal regulation of these signals remain unknown. Here we show that serum IL-5 levels are maintained by long-lived type 2 innate lymphoid cells (ILC2) resident in peripheral tissues. ILC2 cells secrete IL-5 constitutively and are induced to co-express IL-13 during type 2 inflammation, resulting in localized eotaxin production and eosinophil accumulation. In the small intestine where eosinophils and eotaxin are constitutive4, ILC2 cells co-express IL-5 and IL-13; this co-expression is enhanced after caloric intake. The circadian synchronizer vasoactive intestinal peptide also stimulates ILC2 cells through the VPAC2 receptor to release IL-5, linking eosinophil levels with metabolic cycling. Tissue ILC2 cells regulate basal eosinophilopoiesis and tissue eosinophil accumulation through constitutive and stimulated cytokine expression, and this dissociated regulation can be tuned by nutrient intake and central circadian rhythms.
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Rothenberg, M. E. & Hogan, S. P. The eosinophil. Annu. Rev. Immunol. 24, 147–174 (2006)
Takatsu, K. & Nakajima, H. IL-5 and eosinophilia. Curr. Opin. Immunol. 20, 288–294 (2008)
Pope, S. et al. IL-13 induces eosinophil recruitment into the lung by an IL-5- and eotaxin-dependent mechanism. J. Allergy Clin. Immunol. 108, 594–601 (2001)
Mishra, A., Hogan, S., Lee, J., Foster, P. & Rothenberg, M. Fundamental signals that regulate eosinophil homing to the gastrointestinal tract. J. Clin. Invest. 103, 1719–1727 (1999)
Kopf, M. et al. IL-5-deficient mice have a developmental defect in CD5+ B-1 cells and lack eosinophilia but have normal antibody and cytotoxic T cell responses. Immunity 4, 15–24 (1996)
Yoshida, T. et al. Defective B-1 cell development and impaired immunity against Angiostrongylus cantonensis in IL-5Rα-deficient mice. Immunity 4, 483–494 (1996)
Molofsky, A. B. et al. Innate lymphoid type 2 cells sustain visceral adipose tissue eosinophils and alternatively activated macrophages. J. Exp. Med. 210, 535–549 (2013)
Ikutani, M. et al. Identification of innate IL-5-producing cells and their role in lung eosinophil regulation and antitumor immunity. J. Immunol. 188, 703–713 (2012)
Spits, H. & Cupedo, T. Innate lymphoid cells: emerging insights in development, lineage relationships, and function. Annu. Rev. Immunol. 30, 647–675 (2012)
Price, A. et al. Systemically dispersed innate IL-13-expressing cells in type 2 immunity. Proc. Natl Acad. Sci. USA 107, 11489–11494 (2010)
Liang, H. E. et al. Divergent expression patterns of IL-4 and IL-13 define unique functions in allergic immunity. Nature Immunol. 13, 58–66 (2012)
Neill, D. R. et al. Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464, 1367–1370 (2010)
Moro, K. et al. Innate production of TH2 cytokines by adipose tissue-associated c-Kit+Sca-1+ lymphoid cells. Nature 463, 540–544 (2010)
Carlens, J. et al. Common γ-chain-dependent signals confer selective survival of eosinophils in the murine small intestine. J. Immunol. 183, 5600–5607 (2009)
Halberg, F., Visscher, M. & Bittner, J. Eosinophil rhythm in mice: range of occurrence; effects of illumination, feeding, and adrenalectomy. Am. J. Physiol. 174, 109–122 (1953)
Pauly, J. et al. Meal timing dominates the lighting regimen as a synchronizer of the eosinophil rhythm in mice. Acta Anat. 93, 60–68 (1975)
Lelievre, V. et al. Gastrointestinal dysfunction in mice with a targeted mutation in the gene encoding vasoactive intestinal polypeptide: a model for the study of intestinal ileus and Hirschsprung’s disease. Peptides 28, 1688–1699 (2007)
Maywood, E. S. et al. Analysis of core circadian feedback loop in suprachiasmatic nucleus of mCry1-luc transgenic reporter mouse. Proc. Natl Acad. Sci. USA 110, 9547–9552 (2013)
Harmar, A. J. et al. The VPAC2 receptor is essential for circadian function in the mouse suprachiasmatic nuclei. Cell 109, 497–508 (2002)
Colwell, C. S. et al. Disrupted circadian rhythms in VIP- and PHI-deficient mice. Am. J. Physiol. Regul. Integr. Comp. Physiol. 285, R939–R949 (2003)
Sheward, W. J. et al. Entrainment to feeding but not to light: circadian phenotype of VPAC2 receptor-null mice. J. Neurosci. 27, 4351–4358 (2007)
Voice, J. et al. c-Maf and JunB mediation of Th2 differentiation induced by the type 2 G protein-coupled receptor (VPAC2) for vasoactive intestinal peptide. J. Immunol. 172, 7289–7296 (2004)
Samarasinghe, A. E., Hoselton, S. A. & Schuh, J. M. The absence of the VPAC2 receptor does not protect mice from Aspergillus induced allergic asthma. Peptides 31, 1068–1075 (2010)
Tsutsumi, M. et al. A potent and highly selective VPAC2 agonist enhances glucose-induced insulin release and glucose disposal: a potential therapy for type 2 diabetes. Diabetes 51, 1453–1460 (2002)
Dickson, L. & Finlayson, K. VPAC and PAC receptors: From ligands to function. Pharmacol. Ther. 121, 294–316 (2009)
Domarus, A. v. Die bedeutung der kammerzahlung der eosinophilen fur die klinik. Dtsch. Arch. Klin. Med. 171, 333–358 (1931)
Mjösberg, J. M. et al. Human IL-25- and IL-33-responsive type 2 innate lymphoid cells are defined by expression of CRTH2 and CD161. Nature Immunol. 12, 1055–1062 (2011)
Sullivan, B. M. et al. Genetic analysis of basophil function in vivo. Nature Immunol. 12, 527–535 (2011)
Thornton, E. E. et al. Spatiotemporally separated antigen uptake by alveolar dendritic cells and airway presentation to T cells in the lung. J. Exp. Med. 209, 1183–1199 (2012)
Vomhof-DeKrey, E. E. et al. Radical reversal of vasoactive intestinal peptide (VIP) receptors during early lymphopoiesis. Peptides 32, 2058–2066 (2011)
We thank the NIH Tetramer Core Facility for reagents, B. Sullivan, N. Flores, M. Consengco and Z. Wang for technical expertise, and M. Anderson, C. Lowell and M. McCune for comments on the manuscript. Supported by NIH (AI026918, AI030663, AI078869, HL107202), the Diabetes Endocrinology Research Center grant (DK063720), the Howard Hughes Medical Institute and the Sandler Asthma Basic Research Center at the University of California San Francisco. J.C.N. is supported by NIH training grants (AI007641 and AI007334).
The authors declare no competing financial interests.
Extended data figures and tables
a, b, Flow cytometry with intracellular staining for IL-4, IL-13, IL-5 and IFN-γ of CD4+ T cells from wild-type and R5/R5 mice cultured under TH2 (a) or TH1 (b) conditions and then re-stimulated for 24 h. Numbers represent per cent of CD3+CD4+ cells. c, Percentage of cultured CD3+CD4+ cells in the R5+ gate and ELISA for IL-5 from the supernatants in R5/+ and wild-type TH2 cultures re-stimulated on plate-bound anti-CD3ε for 4 days. Data are representative of two independent experiments (a, b), and ELISA data (c) obtained by averaging four replicates per time point.
a, Gating of cells from lung and small intestine. Numbers in first two panels (lung) and first panel (intestine) are percentage of live (DAPI−) cells, and remaining panel previously gated on R5+CD90.2+ cells (lung) or R5+CD45+ cells (intestine). Histograms show R5+CD4−CD5− cells, total CD4+CD5+ cells, and eosinophils (SiglecF+CD11b+SSChi). b, Percentage R5+ of Lin−CD127+T1/ST2+ cells and R5 fluorescence in indicated tissues. c, IL-5 ELISPOT of R5+ cells sorted from the lungs of R5/+ or R5/R5 mice and cultured (3,000 cells per well) for 48 h. Data are representative of two independent experiments with three mice per group (a, b), or representative of two independent experiments using sorted cells pooled from four mice (c). Represented as mean ± s.e.m. Lin, lineage markers (B220, CD5, CD11b, CD11c, Ly6G, FcεRI and NK1.1); MFI, mean fluorescence intensity; *P < 0.05; ***P < 0.001, by Student’s t-test.
a, Flow cytometry of lung cells from mouse strains as indicated. Numbers are percentage of Lin− cells. b, Flow cytometry of lung cells previously gated as in a (Lin−CD90.2+T1/ST2+). Numbers are percentage of Lin−CD90.2+T1/ST2+ cells. c, Total lung ILC2 cells (Lin−CD90.2+T1/ST2+) in mouse strains as indicated. d, Flow cytometry of R5/+ lung; previously gated on Lin−CD90.2+ cells. e, Lung Lin−CD90.2+T1/ST2+ cells as a percentage of CD45+ cells at neonatal day 1, day 8, or week 8 of life. f, Per cent BrdU+ of thymus CD4−CD8− (DN) cells, lung CD4+ T cells, and lung R5+ ILC2 cells after 14 days BrdU. Data are representative of two independent experiments (a, b and d); pooled from three independent experiments for 4 (wild type and IL7rα−/−) or 7 (all others) mice per group (c); pooled from three independent experiments for 5 (day 1), 6 (day 8), or 4 (adult) mice per group (e); or pooled from two independent experiments for 3 mice per group (f). Represented as mean ± s.e.m. Lin, lineage markers (B220, CD5, CD11b, CD11c, Ly6G, FcεRI and NK1.1); NS, not significant by Kruskal–Wallis (c) or by Student’s t-test (f); *P < 0.05; ***P < 0.001, by Student’s t-test.
a, b, Flow cytometry of R5/R5 and R5/R5 ROSA–YFP lungs 12 days after infection with N. brasiliensis, previously gated on live (DAPI−) cells. a, Numbers are per cent YFP+ of live (DAPI−) cells (left two panels), and per cent R5+CD4+ and R5+CD4− of YFP+ cells (second from right). Far right panel, CD90.2 and T1/ST2 staining of R5+CD4− cells. b, Numbers are per cent of CD4+ cells. c, Baseline total cells and per cent CD4+ T cells in bone marrow (single femur), spleen and lung, and CD4+ cells as a per cent of CD45+ cells in small intestine lamina propria of R5/R5 or R5/R5 deleter mice. d, Representative flow cytometry of small intestine lamina propria cells (previously gated as CD45+CD8−NK1.1−). Data are representative of 2 mice in each group from one experiment (a, b); or pooled from three independent experiments for 6 (small intestine, and R5/R5 bone marrow and spleen) or 9 (all others) mice per group (c); or representative of two mice in each group from one experiment (d). Represented as means ± s.e.m. BM, bone marrow; ***P < 0.001 by Student’s t-test.
Flow cytometry and quantification of R5 and KLRG1 fluorescence in ILC2 cells from untreated Rag1−/− and R5/R5 Rag1−/− mice, and R5/R5 Rag1−/− mice treated with IL-2 and IL-33. Lung cells previously gated as Lin−CD90.2+. Data are pooled from two independent experiments for 3 (R5/R5 + PBS) or 8 (R5/R5 + IL-2/IL-33) mice per group. Represented as means ± s.e.m. MFI, mean fluorescence intensity.
a, Schematic for feeding during light-only or dark-only 9 days before collecting tissues. b, Flow cytometry of small intestine cells previously gated on CD45+Lin−CD127+ (left panels), or CD45+Lin−CD127−ICOS+R5+ (right panels) showing human CD4 and R5 fluorescence. c, Per cent R5+ and R5 fluorescence of lamina propria ILC2 cells after 10-day food schedule in a. d, Per cent Smart13+ (S13) of lamina propria after 16-h fast. Data are representative of 2 independent experiments with 4 mice per group (b, c), or one experiment with 4 (fed) or 2 (fasted) mice per group (d). Represented as mean ± s.e.m. Lin, lineage markers (CD4, CD5, CD8, B220, CD11b, CD11c, NK1.1, Gr1); MFI, mean fluorescence intensity.
a, Representative flow cytometry of ILC2 cells from small intestine of wild-type or R5/R5 mice, previously gated on Lin−CD45+KLRG1+. b, c, IL-5 in culture supernatant measured by cytometric bead array. b, Lin−CD45+KLRG1+ ILC2 cells sorted from small intestine cultured at 10,000 per well in IL-7 (10 ng ml−1) alone or with VIP (1 μM) for 6 h. c, Lin−CD90.2+CD25+ ILC2 cells sorted from lung cultured at 5,000 per well in IL-7 (10 ng ml−1) alone or with VIP or VPAC2-specific agonist BAY 55-9837 (both 1 μM) for 18 h. Data are representative of three independent experiments (a) or pooled averages of duplicate cultures from 4 (b) or 3 (c) independent cell sorts. Represented as mean ± s.e.m. Lin, lineage markers (CD4, CD5, CD8, B220, CD11b, CD11c, NK1.1, Gr1); *P < 0.05 by paired Student’s t-test.
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Nussbaum, J., Van Dyken, S., von Moltke, J. et al. Type 2 innate lymphoid cells control eosinophil homeostasis. Nature 502, 245–248 (2013). https://doi.org/10.1038/nature12526
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