Advertisement

Group 2 Innate Lymphoid Cells Coordinate Damage Response in the Stomach

Published:September 03, 2020DOI:https://doi.org/10.1053/j.gastro.2020.08.051

      Background & Aims

      Severe injury to the lining of the stomach leads to changes in the epithelium (reprogramming) that protect and promote repair of the tissue, including development of spasmolytic polypeptide-expressing metaplasia (SPEM) and tuft and foveolar cell hyperplasia. Acute gastric damage elicits a type-2 inflammatory response that includes production of type-2 cytokines and infiltration by eosinophils and alternatively activated macrophages. Stomachs of mice that lack interleukin 33 (IL33) or interleukin 13 (IL13) did not undergo epithelial reprogramming after drug-induced injury. We investigated the role of group 2 innate lymphoid cells (ILC2s) in gastric epithelial repair.

      Methods

      Acute gastric injury was induced in C57BL/6J mice (wild-type and RAG1 knockout) by administration of L635. We isolated ILC2s by flow cytometry from stomachs of mice that were and were not given L635 and performed single-cell RNA sequencing. ILC2s were depleted from wild-type and RAG1-knockout mice by administration of anti-CD90.2. We assessed gastric cell lineages, markers of metaplasia, inflammation, and proliferation. Gastric tissue microarrays from patients with gastric adenocarcinoma were analyzed by immunostaining.

      Results

      There was a significant increase in the number of GATA3-positive ILC2s in stomach tissues from wild-type mice after L635-induced damage, but not in stomach tissues from IL33-knockout mice. We characterized a marker signature of gastric mucosal ILC2s and identified a transcription profile of metaplasia-associated ILC2s, which included changes in expression of Il5, Il13, Csf2, Pd1, and Ramp3; these changes were validated by quantitative polymerase chain reaction and immunocytochemistry. Depletion of ILC2s from mice blocked development of metaplasia after L635-induced injury in wild-type and RAG1-knockout mice and prevented foveolar and tuft cell hyperplasia and infiltration or activation of macrophages after injury. Numbers of ILC2s were increased in stomach tissues from patients with SPEM compared with patients with normal corpus mucosa.

      Conclusions

      In analyses of stomach tissues from mice with gastric tissue damage and patients with SPEM, we found evidence of type 2 inflammation and increased numbers of ILC2s. Our results suggest that ILC2s coordinate the metaplastic response to severe gastric injury.

      Graphical abstract

      Keywords

      Abbreviations used in this paper:

      DMEM (Dulbecco’s modified Eagle’s medium), FACS (fluorescence-activated cell sorter), GIF (gastric intrinsic factor), Ig (immunoglobulin), IL (interleukin), ILCs (innate lymphoid cells), PBS (phosphate-buffered saline), SPEM (spasmolytic polypeptide-expressing metaplasia), TMA (tissue microarrays)
      To read this article in full you will need to make a payment
      AGA Member Login
      Login with your AGA username and password.
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Petersen C.P.
        • Meyer A.R.
        • De Salvo C.
        • et al.
        A signalling cascade of IL-33 to IL-13 regulates metaplasia in the mouse stomach.
        Gut. 2017; 67: 805-817
        • Schwartz C.
        • O'Grady K.
        • Lavelle E.C.
        • et al.
        Interleukin 33: an innate alarm for adaptive responses beyond Th2 immunity-emerging roles in obesity, intestinal inflammation, and cancer.
        Eur J Immunol. 2016; 46: 1091-1100
        • Buzzelli J.N.
        • Chalinor H.V.
        • Pavlic D.I.
        • et al.
        IL33 is a stomach alarmin that initiates a skewed Th2 response to injury and infection.
        Cell Mol Gastroenterol Hepatol. 2015; 1: 203-221.e3
        • Neill D.R.
        • Wong S.H.
        • Bellosi A.
        • et al.
        Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity.
        Nature. 2010; 464: 1367-1370
        • Guo L.
        • Junttila I.S.
        • Paul W.E.
        Cytokine-induced cytokine production by conventional and innate lymphoid cells.
        Trends Immunol. 2012; 33: 598-606
        • Eberl G.
        • Colonna M.
        • Di Santo J.P.
        • et al.
        Innate lymphoid cells. Innate lymphoid cells: a new paradigm in immunology.
        Science. 2015; 348aaa6566
        • Spits H.
        • Di Santo J.P.
        The expanding family of innate lymphoid cells: regulators and effectors of immunity and tissue remodeling.
        Nat Immunol. 2011; 12: 21-27
        • Serafini N.
        • Vosshenrich C.A.
        • Di Santo J.P.
        Transcriptional regulation of innate lymphoid cell fate.
        Nat Rev Immunol. 2015; 15: 415-428
        • Klein Wolterink R.G.
        • Serafini N.
        • van Nimwegen M.
        • et al.
        Essential, dose-dependent role for the transcription factor Gata3 in the development of IL-5+ and IL-13+ type 2 innate lymphoid cells.
        Proc Natl Acad Sci U S A. 2013; 110: 10240-10245
        • Li D.
        • Guabiraba R.
        • Besnard A.G.
        • et al.
        IL-33 promotes ST2-dependent lung fibrosis by the induction of alternatively activated macrophages and innate lymphoid cells in mice.
        J Allergy Clin Immunol. 2014; 134: 1422-1432.e11
        • Salimi M.
        • Barlow J.L.
        • Saunders S.P.
        • et al.
        A role for IL-25 and IL-33-driven type-2 innate lymphoid cells in atopic dermatitis.
        J Exp Med. 2013; 210: 2939-2950
        • Stier M.T.
        • Bloodworth M.H.
        • Toki S.
        • et al.
        Respiratory syncytial virus infection activates IL-13-producing group 2 innate lymphoid cells through thymic stromal lymphopoietin.
        J Allergy Clin Immunol. 2016; 138: 814-824.e11
        • Fuchs A.
        • Colonna M.
        Innate lymphoid cells in homeostasis, infection, chronic inflammation and tumors of the gastrointestinal tract.
        Curr Opin Gastroenterol. 2013; 29: 581-587
        • Satoh-Takayama N.
        • Kato T.
        • Motomura Y.
        • et al.
        Bacteria-induced group 2 innate lymphoid cells in the stomach provide immune protection through induction of IgA.
        Immunity. 2020; 52: 635-649
        • Bernink J.H.
        • Germar K.
        • Spits H.
        The role of ILC2 in pathology of type 2 inflammatory diseases.
        Curr Opin Immunol. 2014; 31: 115-120
        • Goldenring J.R.
        Pyloric metaplasia, pseudopyloric metaplasia, ulcer-associated cell lineage and spasmolytic polypeptide-expressing metaplasia: reparative lineages in the gastrointestinal mucosa.
        J Pathol. 2018; 245: 132-137
        • Burkitt M.D.
        • Duckworth C.A.
        • Williams J.M.
        • et al.
        Helicobacter pylori-induced gastric pathology: insights from in vivo and ex vivo models.
        Dis Model Mech. 2017; 10: 89-104
        • Nam K.T.
        • Lee H.J.
        • Sousa J.F.
        • et al.
        Mature chief cells are cryptic progenitors for metaplasia in the stomach.
        Gastroenterology. 2010; 139: 2028-2037.e9
        • Barlow J.L.
        • Bellosi A.
        • Hardman C.S.
        • et al.
        Innate IL-13-producing nuocytes arise during allergic lung inflammation and contribute to airways hyperreactivity.
        J Allergy Clin Immunol. 2012; 129: 191-198.e1–e4
        • Petersen C.P.
        • Weis V.G.
        • Nam K.T.
        • et al.
        Macrophages promote progression of spasmolytic polypeptide-expressing metaplasia after acute loss of parietal cells.
        Gastroenterology. 2014; 146: 1727-1738.e8
        • Leys C.M.
        • Nomura S.
        • Rudzinski E.
        • et al.
        Expression of Pdx-1 in human gastric metaplasia and gastric adenocarcinoma.
        Hum Pathol. 2006; 37: 1162-1168
        • Weis V.G.
        • Petersen C.P.
        • Weis J.A.
        • et al.
        Maturity and age influence chief cell ability to transdifferentiate into metaplasia.
        Am J Physiol Gastrointest Liver Physiol. 2017; 312: G67-G76
        • Jones T.R.
        • Kang I.H.
        • Wheeler D.B.
        • et al.
        CellProfiler Analyst: data exploration and analysis software for complex image-based screens.
        BMC Bioinformatics. 2008; 9: 482
        • Tait Wojno E.D.
        • Beamer C.A.
        Isolation and identification of innate lymphoid cells (ILCs) for immunotoxicity testing.
        Methods Mol Biol. 2018; 1803: 353-370
        • Hoyler T.
        • Klose C.S.
        • Souabni A.
        • et al.
        The transcription factor GATA-3 controls cell fate and maintenance of type 2 innate lymphoid cells.
        Immunity. 2012; 37: 634-648
        • Picelli S.
        • Faridani O.R.
        • Bjorklund A.K.
        • et al.
        Full-length RNA-seq from single cells using Smart-seq2.
        Nat Protoc. 2014; 9: 171-181
        • Gour N.
        • Smole U.
        • Yong H.M.
        • et al.
        C3a is required for ILC2 function in allergic airway inflammation.
        Mucosal Immunol. 2018; 11: 1653-1662
        • Robinette M.L.
        • Fuchs A.
        • Cortez V.S.
        • et al.
        Transcriptional programs define molecular characteristics of innate lymphoid cell classes and subsets.
        Nat Immunol. 2015; 16: 306-317
        • Lei A.H.
        • Xiao Q.
        • Liu G.Y.
        • et al.
        ICAM-1 controls development and function of ILC2.
        J Exp Med. 2018; 215: 2157-2174
        • Moro K.
        • Yamada T.
        • Tanabe M.
        • et al.
        Innate production of T(H)2 cytokines by adipose tissue-associated c-Kit(+)Sca-1(+) lymphoid cells.
        Nature. 2010; 463: 540-544
        • Kim B.S.
        • Wang K.
        • Siracusa M.C.
        • et al.
        Basophils promote innate lymphoid cell responses in inflamed skin.
        J Immunol. 2014; 193: 3717-3725
        • Taylor S.
        • Huang Y.
        • Mallett G.
        • et al.
        PD-1 regulates KLRG1(+) group 2 innate lymphoid cells.
        J Exp Med. 2017; 214: 1663-1678
        • Sui P.
        • Wiesner D.L.
        • Xu J.
        • et al.
        Pulmonary neuroendocrine cells amplify allergic asthma responses.
        Science. 2018; 360 (eaan3546)
        • Nagashima H.
        • Mahlakoiv T.
        • Shih H.Y.
        • et al.
        Neuropeptide CGRP Limits group 2 innate lymphoid cell responses and constrains type 2 inflammation.
        immunity. 2019; 51: 682-695 e6
        • Wallrapp A.
        • Burkett P.R.
        • Riesenfeld S.J.
        • et al.
        Calcitonin gene-related peptide negatively regulates alarmin-driven type 2 innate lymphoid cell responses.
        Immunity. 2019; 51: 709-723.e6
        • Hayakawa Y.
        • Ariyama H.
        • Stancikova J.
        • et al.
        Mist1 Expressing gastric stem cells maintain the normal and neoplastic gastric epithelium and are supported by a perivascular stem cell niche.
        Cancer Cell. 2015; 28: 800-814
        • Engelbertsen D.
        • Foks A.C.
        • Alberts-Grill N.
        • et al.
        Expansion of CD25+ innate lymphoid cells reduces atherosclerosis.
        Arterioscler Thromb Vasc Biol. 2015; 35: 2526-2535
        • Monticelli L.A.
        • Sonnenberg G.F.
        • Abt M.C.
        • et al.
        Innate lymphoid cells promote lung-tissue homeostasis after infection with influenza virus.
        Nat Immunol. 2011; 12: 1045-1054
        • Ramsey V.G.
        • Doherty J.M.
        • Chen C.C.
        • et al.
        The maturation of mucus-secreting gastric epithelial progenitors into digestive-enzyme secreting zymogenic cells requires Mist1.
        Development. 2007; 134: 211-222
        • Saqui-Salces M.
        • Keeley T.M.
        • Grosse A.S.
        • et al.
        Gastric tuft cells express DCLK1 and are expanded in hyperplasia.
        Histochem Cell Biol. 2011; 136: 191-204
        • Gerbe F.
        • Legraverend C.
        • Jay P.
        The intestinal epithelium tuft cells: specification and function.
        Cell Mol Life Sci. 2012; 69: 2907-2917
        • Choi E.
        • Petersen C.P.
        • Lapierre L.A.
        • et al.
        Dynamic expansion of gastric mucosal doublecortin-like kinase 1-expressing cells in response to parietal cell loss is regulated by gastrin.
        Am J Pathol. 2015; 185: 2219-2231
        • Howitt M.R.
        • Lavoie S.
        • Michaud M.
        • et al.
        Tuft cells, taste-chemosensory cells, orchestrate parasite type 2 immunity in the gut.
        Science. 2016; 351: 1329-1333
        • von Moltke J.
        • Ji M.
        • Liang H.E.
        • et al.
        Tuft-cell-derived IL-25 regulates an intestinal ILC2-epithelial response circuit.
        Nature. 2016; 529: 221-225
        • Leushacke M.
        • Tan S.H.
        • Wong A.
        • et al.
        Lgr5-expressing chief cells drive epithelial regeneration and cancer in the oxyntic stomach.
        Nat Cell Biol. 2017; 19: 774-786
        • Radyk M.D.
        • Burclaff J.
        • Willet S.G.
        • et al.
        Metaplastic cells in the stomach arise, independently of stem cells, via dedifferentiation or transdifferentiation of chief cells.
        Gastroenterology. 2018; 154: 839-843
        • Nomura S.
        • Yamaguchi H.
        • Ogawa M.
        • et al.
        Alterations in gastric mucosal lineages induced by acute oxyntic atrophy in wild-type and gastrin-deficient mice.
        Am J Physiol Gastrointest Liver Physiol. 2005; 288: G362-G375
        • Weller P.F.
        • Spencer L.A.
        Functions of tissue-resident eosinophils.
        Nat Rev Immunol. 2017; 17: 746-760
        • Engevik A.C.
        • Feng R.
        • Choi E.
        • et al.
        The development of spasmolytic polypeptide/TFF2-expressing metaplasia (SPEM) during gastric repair is absent in the aged stomach.
        Cell Mol Gastroenterol Hepatol. 2016; 2: 605-624
        • Scanlon S.T.
        • McKenzie A.N.
        Type 2 innate lymphoid cells: new players in asthma and allergy.
        Curr Opin Immunol. 2012; 24: 707-712
        • Ricardo-Gonzalez R.R.
        • Van Dyken S.J.
        • Schneider C.
        • et al.
        Tissue signals imprint ILC2 identity with anticipatory function.
        Nat Immunol. 2018; 19: 1093-1099