Altered Mucosal Microbiome Diversity and Disease Severity in Sjögren Syndrome

There is mounting evidence that the microbiome has potent immunoregulatory functions. We assessed the effects of intestinal dysbiosis in a model of Sjögren syndrome (SS) by subjecting mice to desiccating stress (DS) and antibiotics (ABX). We characterized the conjunctival, tongue and fecal microbiome profiles of patients with SS. Severity of ocular surface and systemic disease was graded. 16S ribosomal RNA gene sequencing characterized the microbiota. ABX + DS mice had a significantly worse dry eye phenotype compared to controls, a decrease in Clostridium and an increase in Enterobacter, Escherichia/Shigella, and Pseudomonas in stool after ABX + DS for 10 days. Goblet cell density was significantly lower in ABX treated groups compared to controls. Stool from SS subjects had greater relative abundances of Pseudobutyrivibrio, Escherichia/Shigella, Blautia, and Streptococcus, while relative abundance of Bacteroides, Parabacteroides, Faecalibacterium, and Prevotella was reduced compared to controls. The severity of SS ocular and systemic disease was inversely correlated with microbial diversity. These findings suggest that SS is marked by a dysbiotic intestinal microbiome driven by low relative abundance of commensal bacteria and high relative abundance of potentially pathogenic genera that is associated with worse ocular mucosal disease in a mouse model of SS and in SS patients.

[1]  S. Pflugfelder,et al.  Interferon-γ-Induced Unfolded Protein Response in Conjunctival Goblet Cells as a Cause of Mucin Deficiency in Sjögren Syndrome. , 2016, The American journal of pathology.

[2]  Katherine H. Huang,et al.  Structure, Function and Diversity of the Healthy Human Microbiome , 2012, Nature.

[3]  De-Quan Li,et al.  Corticosteroid and doxycycline suppress MMP-9 and inflammatory cytokine expression, MAPK activation in the corneal epithelium in experimental dry eye. , 2006, Experimental eye research.

[4]  R. V. Van Gelder,et al.  Considerations in understanding the ocular surface microbiome. , 2014, American journal of ophthalmology.

[5]  De-Quan Li,et al.  Entrapment of conjunctival goblet cells by desiccation-induced cornification. , 2011, Investigative ophthalmology & visual science.

[6]  A. Peck,et al.  Unraveling the pathophysiology of Sjogren syndrome-associated dry eye disease. , 2009, The ocular surface.

[7]  A. C. Johannessen,et al.  Estimation of the prevalence of primary Sjögren's syndrome in two age‐different community‐based populations using two sets of classification criteria: the Hordaland Health Study , 2008, Scandinavian journal of rheumatology.

[8]  C. Bernstein,et al.  Association Between the Use of Antibiotics in the First Year of Life and Pediatric Inflammatory Bowel Disease , 2010, The American Journal of Gastroenterology.

[9]  R. Xavier,et al.  Regulation of inflammatory responses by gut microbiota and chemoattractant receptor GPR43 , 2009, Nature.

[10]  P. Asbell,et al.  Clinical guidelines for management of dry eye associated with Sjögren disease. , 2015, The ocular surface.

[11]  E. Veerman,et al.  Antimicrobial defense systems in saliva. , 2014, Monographs in oral science.

[12]  William A. Walters,et al.  Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms , 2012, The ISME Journal.

[13]  A. Tzioufas,et al.  Pathogenesis of Sjögren's syndrome: what we know and what we should learn. , 2012, Journal of autoimmunity.

[14]  R. Dana,et al.  Amelioration of murine dry eye disease by topical antagonist to chemokine receptor 2. , 2009, Archives of ophthalmology.

[15]  I. van Seuningen,et al.  Resistin‐like molecule &bgr; regulates intestinal mucous secretion and curtails TNBS‐induced colitis in mice , 2008, Inflammatory bowel diseases.

[16]  R. Dana,et al.  A novel pro-lymphangiogenic function for Th17/IL-17. , 2011, Blood.

[17]  J. Anaya,et al.  Genetic Basis of Sjögren's Syndrome. How Strong is the Evidence? , 2006, Clinical & developmental immunology.

[18]  Fiona Powrie,et al.  Microbiota, Disease, and Back to Health: A Metastable Journey , 2012, Science Translational Medicine.

[19]  E. A. Volpe,et al.  Aqueous Tear Deficiency Increases Conjunctival Interferon-γ (IFN-γ) Expression and Goblet Cell Loss. , 2015, Investigative ophthalmology & visual science.

[20]  Katherine H. Huang,et al.  A framework for human microbiome research , 2012, Nature.

[21]  J. Doyle,et al.  Utility assessment to measure the impact of dry eye disease. , 2006, The ocular surface.

[22]  R. Dana,et al.  Modulation of integrin alpha4beta1 (VLA-4) in dry eye disease. , 2008, Archives of ophthalmology.

[23]  De-Quan Li,et al.  Dry eye-induced conjunctival epithelial squamous metaplasia is modulated by interferon-gamma. , 2007, Investigative ophthalmology & visual science.

[24]  H. Sokol,et al.  Faecalibacterium prausnitzii and human intestinal health. , 2013, Current opinion in microbiology.

[25]  W. Feuer,et al.  Conjunctival Cytologiec Features of Primary Sjögren's Syndrome , 1990 .

[26]  L. Criswell,et al.  American College of Rheumatology classification criteria for Sjögren's syndrome: A data‐driven, expert consensus approach in the Sjögren's International Collaborative Clinical Alliance Cohort , 2012, Arthritis care & research.

[27]  J. Doré,et al.  Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients , 2008, Proceedings of the National Academy of Sciences.

[28]  A. Rudensky,et al.  Metabolites produced by commensal bacteria promote peripheral regulatory T cell generation , 2013, Nature.

[29]  W. Feuer,et al.  Conjunctival cytologic features of primary Sjögren's syndrome. , 1990, Ophthalmology.

[30]  R. Jonsson,et al.  Clinical, immunological, and immunogenetic aspects of autoantibody production against Ro/SSA, La/SSB and their linear epitopes in primary Sjögren's syndrome (pSS): a European multicentre study , 2002, Annals of the rheumatic diseases.

[31]  E. A. Volpe,et al.  Topical interferon-gamma neutralization prevents conjunctival goblet cell loss in experimental murine dry eye. , 2014, Experimental eye research.

[32]  D. Corry,et al.  Homeostatic control of conjunctival mucosal goblet cells by NKT-derived IL-13 , 2011, Mucosal Immunology.

[33]  B. B. Finlay,et al.  Effects of antibiotics on human microbiota and subsequent disease. , 2014, Annual review of microbiology.

[34]  K. Honda,et al.  Induction of Colonic Regulatory T Cells by Indigenous Clostridium Species , 2011, Science.

[35]  H. Dua,et al.  Antimicrobial defensin peptides of the human ocular surface , 1999, The British journal of ophthalmology.

[36]  Pelin Yilmaz,et al.  The SILVA ribosomal RNA gene database project: improved data processing and web-based tools , 2012, Nucleic Acids Res..

[37]  Shital Patel,et al.  The Human Microbiome Project strategy for comprehensive sampling of the human microbiome and why it matters , 2013, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[38]  K. Berer,et al.  Commensal microbiota and myelin autoantigen cooperate to trigger autoimmune demyelination , 2011, Nature.

[39]  K. Honda,et al.  Microbiota-Dependent Activation of an Autoreactive T Cell Receptor Provokes Autoimmunity in an Immunologically Privileged Site. , 2015, Immunity.

[40]  H. Moutsopoulos,et al.  Characteristics of the minor salivary gland infiltrates in Sjögren's syndrome. , 2010, Journal of autoimmunity.

[41]  C. Schwartz,et al.  Resolvin E1 (RX-10001) Reduces Corneal Epithelial Barrier Disruption and Protects Against Goblet Cell Loss in a Murine Model of Dry Eye , 2012, Cornea.

[42]  T. Macfarlane,et al.  Changes in the oral flora in Sjögren's syndrome , 1974, Journal of clinical pathology.

[43]  F. Bushman,et al.  Commensal bacterial–derived signals regulate basophil hematopoiesis and allergic inflammation , 2012, Nature Medicine.

[44]  D. Corry,et al.  IL-13 Stimulates Proliferation and Expression of Mucin and Immunomodulatory Genes in Cultured Conjunctival Goblet Cells. , 2015, Investigative ophthalmology & visual science.

[45]  A. Murphy,et al.  Goblet Cell-Derived Resistin-Like Molecule β Augments CD4+ T Cell Production of IFN-γ and Infection-Induced Intestinal Inflammation1 , 2008, The Journal of Immunology.

[46]  G. Baron,et al.  EULAR Sjögren's syndrome disease activity index: development of a consensus systemic disease activity index for primary Sjögren's syndrome , 2009, Annals of the rheumatic diseases.

[47]  V. Young,et al.  Microbial ecology of the murine gut associated with the development of dextran sodium sulfate‐induced colitis , 2011, Inflammatory bowel diseases.

[48]  R. Ley,et al.  Innate immunity and intestinal microbiota in the development of Type 1 diabetes , 2008, Nature.

[49]  De-Quan Li,et al.  Apical corneal barrier disruption in experimental murine dry eye is abrogated by methylprednisolone and doxycycline. , 2006, Investigative ophthalmology & visual science.

[50]  A. Murphy,et al.  Goblet cell-derived resistin-like molecule beta augments CD4+ T cell production of IFN-gamma and infection-induced intestinal inflammation. , 2008, Journal of immunology.

[51]  L. Bielory,et al.  Pharmacoeconomics of anterior ocular inflammatory disease , 2013, Current opinion in allergy and clinical immunology.

[52]  S. Pflugfelder,et al.  Altered cytokine balance in the tear fluid and conjunctiva of patients with Sjögren's syndrome keratoconjunctivitis sicca. , 1999, Current eye research.

[53]  C. Schaumburg,et al.  Dry Eye as a Mucosal Autoimmune Disease , 2013, International reviews of immunology.

[54]  L. Criswell,et al.  Differentiation of follicular helper T cells by salivary gland epithelial cells in primary Sjögren's syndrome. , 2014, Journal of autoimmunity.

[55]  Leah M. Feazel,et al.  Sex Differences in the Gut Microbiome Drive Hormone-Dependent Regulation of Autoimmunity , 2013, Science.

[56]  De-Quan Li,et al.  Production and activity of matrix metalloproteinase-9 on the ocular surface increase in dysfunctional tear syndrome. , 2009, Investigative ophthalmology & visual science.

[57]  Robert C. Edgar,et al.  UPARSE: highly accurate OTU sequences from microbial amplicon reads , 2013, Nature Methods.

[58]  S. Pflugfelder,et al.  Desiccating Stress Decreases Apical Corneal Epithelial Cell Size-Modulation by the Metalloproteinase Inhibitor Doxycycline , 2008, Cornea.

[59]  R. Dana,et al.  Therapeutic Efficacy of Topical Epigallocatechin Gallate in Murine Dry Eye , 2011, Cornea.

[60]  R. Knight,et al.  UniFrac: a New Phylogenetic Method for Comparing Microbial Communities , 2005, Applied and Environmental Microbiology.

[61]  S. Carsons.  A review and update of Sjögren's syndrome: manifestations, diagnosis, and treatment. , 2001, The American journal of managed care.

[62]  De-Quan Li,et al.  IL-17 disrupts corneal barrier following desiccating stress , 2009, Mucosal Immunology.

[63]  E. A. Volpe,et al.  Interferon- (cid:1) Exacerbates Dry Eye–Induced Apoptosis in Conjunctiva through Dual Apoptotic Pathways , 2022 .

[64]  M. Tomita,et al.  Commensal microbe-derived butyrate induces the differentiation of colonic regulatory T cells , 2013, Nature.

[65]  R. Dana,et al.  The CCR6/CCL20 axis mediates Th17 cell migration to the ocular surface in dry eye disease. , 2013, Investigative ophthalmology & visual science.