The Microbiota Mediates Pathogen Clearance from the Gut Lumen after Non-Typhoidal Salmonella Diarrhea

Many enteropathogenic bacteria target the mammalian gut. The mechanisms protecting the host from infection are poorly understood. We have studied the protective functions of secretory antibodies (sIgA) and the microbiota, using a mouse model for S. typhimurium diarrhea. This pathogen is a common cause of diarrhea in humans world-wide. S. typhimurium (S. tm att, sseD) causes a self-limiting gut infection in streptomycin-treated mice. After 40 days, all animals had overcome the disease, developed a sIgA response, and most had cleared the pathogen from the gut lumen. sIgA limited pathogen access to the mucosal surface and protected from gut inflammation in challenge infections. This protection was O-antigen specific, as demonstrated with pathogens lacking the S. typhimurium O-antigen (wbaP, S. enteritidis) and sIgA-deficient mice (TCRβ−/−δ−/−, JH −/−, IgA−/−, pIgR−/−). Surprisingly, sIgA-deficiency did not affect the kinetics of pathogen clearance from the gut lumen. Instead, this was mediated by the microbiota. This was confirmed using ‘L-mice’ which harbor a low complexity gut flora, lack colonization resistance and develop a normal sIgA response, but fail to clear S. tm att from the gut lumen. In these mice, pathogen clearance was achieved by transferring a normal complex microbiota. Thus, besides colonization resistance ( = pathogen blockage by an intact microbiota), the microbiota mediates a second, novel protective function, i.e. pathogen clearance. Here, the normal microbiota re-grows from a state of depletion and disturbed composition and gradually clears even very high pathogen loads from the gut lumen, a site inaccessible to most “classical” immune effector mechanisms. In conclusion, sIgA and microbiota serve complementary protective functions. The microbiota confers colonization resistance and mediates pathogen clearance in primary infections, while sIgA protects from disease if the host re-encounters the same pathogen. This has implications for curing S. typhimurium diarrhea and for preventing transmission.

[1]  C. von Mering,et al.  Like Will to Like: Abundances of Closely Related Species Can Predict Susceptibility to Intestinal Colonization by Pathogenic and Commensal Bacteria , 2010, PLoS pathogens.

[2]  V. Kunin,et al.  Wrinkles in the rare biosphere: pyrosequencing errors can lead to artificial inflation of diversity estimates. , 2009, Environmental microbiology.

[3]  P. Brandtzaeg.  Mucosal Immunity: Induction, Dissemination, and Effector Functions , 2009, Scandinavian journal of immunology.

[4]  Dan R. Littman,et al.  Induction of Intestinal Th17 Cells by Segmented Filamentous Bacteria , 2009, Cell.

[5]  Annaïg Lan,et al.  The key role of segmented filamentous bacteria in the coordinated maturation of gut helper T cell responses. , 2009, Immunity.

[6]  L. Hooper,et al.  Immune responses to the microbiota at the intestinal mucosal surface. , 2009, Immunity.

[7]  C. Quince,et al.  Accurate determination of microbial diversity from 454 pyrosequencing data , 2009, Nature Methods.

[8]  Thomas F. Tedder,et al.  Innate and Adaptive Immunity Cooperate Flexibly to Maintain Host-Microbiota Mutualism , 2009, Science.

[9]  P. Teunis,et al.  Estimation of incidences of infectious diseases based on antibody measurements , 2009, Statistics in medicine.

[10]  Balamurugan Periaswamy,et al.  Accelerated Type III Secretion System 2-Dependent Enteropathogenesis by a Salmonella enterica Serovar Enteritidis PT4/6 Strain , 2009, Infection and Immunity.

[11]  M. Aebi,et al.  O-Antigen-Negative Salmonella enterica Serovar Typhimurium Is Attenuated in Intestinal Colonization but Elicits Colitis in Streptomycin-Treated Mice , 2009, Infection and Immunity.

[12]  James R. Cole,et al.  The Ribosomal Database Project: improved alignments and new tools for rRNA analysis , 2008, Nucleic Acids Res..

[13]  B. Roe,et al.  A core gut microbiome in obese and lean twins , 2008, Nature.

[14]  Christian von Mering,et al.  STRING 8—a global view on proteins and their functional interactions in 630 organisms , 2008, Nucleic Acids Res..

[15]  Susan M. Huse,et al.  Exploring Microbial Diversity and Taxonomy Using SSU rRNA Hypervariable Tag Sequencing , 2008, PLoS genetics.

[16]  Les Dethlefsen,et al.  The Pervasive Effects of an Antibiotic on the Human Gut Microbiota, as Revealed by Deep 16S rRNA Sequencing , 2008, PLoS biology.

[17]  G. Brysbaert,et al.  EBER2 RNA-induced transcriptome changes identify cellular processes likely targeted during Epstein Barr Virus infection , 2008, BMC Research Notes.

[18]  R. Knight,et al.  Evolution of Mammals and Their Gut Microbes , 2008, Science.

[19]  W. Rabsch,et al.  Motility allows S. Typhimurium to benefit from the mucosal defence , 2008, Cellular microbiology.

[20]  Wolf-Dietrich Hardt,et al.  The role of microbiota in infectious disease. , 2008, Trends in microbiology.

[21]  Steffen Jung,et al.  Microbe sampling by mucosal dendritic cells is a discrete, MyD88-independent stepin ΔinvG S. Typhimurium colitis , 2008, The Journal of experimental medicine.

[22]  N. Salzman,et al.  Enteric Salmonellosis Disrupts the Microbial Ecology of the Murine Gastrointestinal Tract , 2007, Infection and Immunity.

[23]  M. Rescigno,et al.  Entry route of Salmonella typhimurium directs the type of induced immune response. , 2007, Immunity.

[24]  G. Dougan,et al.  Salmonella enterica Serovar Typhimurium Exploits Inflammation to Compete with the Intestinal Microbiota , 2007, PLoS biology.

[25]  J. Rawls.  Enteric infection and inflammation alter gut microbial ecology. , 2007, Cell host & microbe.

[26]  Barry S. Michaels,et al.  Outbreaks where food workers have been implicated in the spread of foodborne disease. Part 1. Description of the problem, methods, and agents involved. , 2007, Journal of food protection.

[27]  J. Tiedje,et al.  Naïve Bayesian Classifier for Rapid Assignment of rRNA Sequences into the New Bacterial Taxonomy , 2007, Applied and Environmental Microbiology.

[28]  Sebastian Noth,et al.  Normalization Using Weighted Negative Second Order Exponential Error Functions (NeONORM) Provides Robustness Against Asymmetries in Comparative Transcriptome Profiles and Avoids False Calls , 2006, Genom. Proteom. Bioinform..

[29]  Susan M. Huse,et al.  Microbial diversity in the deep sea and the underexplored “rare biosphere” , 2006, Proceedings of the National Academy of Sciences.

[30]  Eoin L. Brodie,et al.  Greengenes, a Chimera-Checked 16S rRNA Gene Database and Workbench Compatible with ARB , 2006, Applied and Environmental Microbiology.

[31]  R. Strugnell,et al.  Innate secretory antibodies protect against natural Salmonella typhimurium infection , 2006, The Journal of experimental medicine.

[32]  S. Hapfelmeier,et al.  Virulence of Broad- and Narrow-Host-Range Salmonella enterica Serovars in the Streptomycin-PretreatedMouse Model , 2006, Infection and Immunity.

[33]  B. Finlay,et al.  Analysis of the Contribution of Salmonella Pathogenicity Islands 1 and 2 to Enteric Disease Progression Using a Novel Bovine Ileal Loop Model and a Murine Model of Infectious Enterocolitis , 2005, Infection and Immunity.

[34]  Rachel E. Klevit,et al.  Recognition of Antimicrobial Peptides by a Bacterial Sensor Kinase , 2005, Cell.

[35]  F. Bäckhed,et al.  Host-Bacterial Mutualism in the Human Intestine , 2005, Science.

[36]  A. Aderem,et al.  CD4+ T Cells and Toll-Like Receptors Recognize Salmonella Antigens Expressed in Bacterial Surface Organelles , 2005, Infection and Immunity.

[37]  S. Akira,et al.  The Salmonella Pathogenicity Island (SPI)-2 and SPI-1 Type III Secretion Systems Allow Salmonella Serovar typhimurium to Trigger Colitis via MyD88-Dependent and MyD88-Independent Mechanisms1 , 2005, The Journal of Immunology.

[38]  C. Simmons,et al.  Vaccine‐induced protection against gastrointestinal bacterial infections in the absence of secretory antibodies , 2005, European journal of immunology.

[39]  S. Hapfelmeier,et al.  A mouse model for S. typhimurium-induced enterocolitis. , 2005, Trends in microbiology.

[40]  B. Stecher,et al.  Flagella and Chemotaxis Are Required for Efficient Induction of Salmonella enterica Serovar Typhimurium Colitis in Streptomycin-Pretreated Mice , 2004, Infection and Immunity.

[41]  B. Finlay,et al.  Clearance of Citrobacter rodentium Requires B Cells but Not Secretory Immunoglobulin A (IgA) or IgM Antibodies , 2004, Infection and Immunity.

[42]  P. Michel,et al.  Risk factors for Salmonella Typhimurium DT104 and non-DT104 infection: a Canadian multi-provincial case-control study , 2004, Epidemiology and Infection.

[43]  J. Fox,et al.  Spatial Distribution and Stability of the Eight Microbial Species of the Altered Schaedler Flora in the Mouse Gastrointestinal Tract , 2004, Applied and Environmental Microbiology.

[44]  B. Stecher,et al.  Role of the Salmonella Pathogenicity Island 1 Effector Proteins SipA, SopB, SopE, and SopE2 in Salmonella enterica Subspecies 1 Serovar Typhimurium Colitis in Streptomycin-Pretreated Mice , 2004, Infection and Immunity.

[45]  M. Hogardt,et al.  Pretreatment of Mice with Streptomycin Provides a Salmonella enterica Serovar Typhimurium Colitis Model That Allows Analysis of Both Pathogen and Host , 2003, Infection and Immunity.

[46]  R. Strugnell,et al.  Role of the Polymeric Ig Receptor in Mucosal B Cell Homeostasis1 , 2003, The Journal of Immunology.

[47]  Takahiro Kanagawa,et al.  Bias and artifacts in multitemplate polymerase chain reactions (PCR). , 2003, Journal of bioscience and bioengineering.

[48]  Sean R. Eddy,et al.  A memory-efficient dynamic programming algorithm for optimal alignment of a sequence to an RNA secondary structure , 2002, BMC Bioinformatics.

[49]  R. Curtiss,et al.  Immune Responses to Recombinant Pneumococcal PspA Antigen Delivered by Live Attenuated Salmonella enterica Serovar Typhimurium Vaccine , 2002, Infection and Immunity.

[50]  C. Mueller.  Tumour necrosis factor in mouse models of chronic intestinal inflammation , 2002, Immunology.

[51]  J. Hughes,et al.  Counting the Uncountable: Statistical Approaches to Estimating Microbial Diversity , 2001, Applied and Environmental Microbiology.

[52]  E. L. Hohmann.  Nontyphoidal salmonellosis. , 2001, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[53]  R. Zinkernagel,et al.  A primitive T cell-independent mechanism of intestinal mucosal IgA responses to commensal bacteria. , 2000, Science.

[54]  E. Hohmann,et al.  Pilot Study of phoP/phoQ-DeletedSalmonella enterica Serovar Typhimurium ExpressingHelicobacter pylori Urease in Adult Volunteers , 2000, Infection and Immunity.

[55]  S. Kaufmann,et al.  Immune response to infection with Salmonella typhimurium in mice , 2000, Journal of leukocyte biology.

[56]  S. Kaufmann,et al.  Cutting Edge: Role of B Lymphocytes in Protective Immunity Against Salmonella typhimurium Infection , 2000, The Journal of Immunology.

[57]  P. Krajči,et al.  Absence of Epithelial Immunoglobulin a Transport, with Increased Mucosal Leakiness, in Polymeric Immunoglobulin Receptor/Secretory Component–Deficient Mice , 1999, The Journal of experimental medicine.

[58]  A. Bradley,et al.  Targeted deletion of the IgA constant region in mice leads to IgA deficiency with alterations in expression of other Ig isotypes. , 1999, Journal of immunology.

[59]  S. Ahmed,et al.  Convalescent excretion of Salmonella enteritidis in infants. , 1999, The Journal of infection.

[60]  S. Miller,et al.  PmrA–PmrB‐regulated genes necessary for 4‐aminoarabinose lipid A modification and polymyxin resistance , 1998, Molecular microbiology.

[61]  H. Ljunggren,et al.  Peripheral lymphoid development and function in TCR mutant mice. , 1994, International immunology.

[62]  E. Vollaard,et al.  Colonization resistance , 1994, Antimicrobial Agents and Chemotherapy.

[63]  R. Curtiss,et al.  Live oral avirulent Salmonella vaccines. , 1993, Veterinary microbiology.

[64]  F. Alt,et al.  Immunoglobulin gene rearrangement in B cell deficient mice generated by targeted deletion of the JH locus. , 1993, International immunology.

[65]  R. F. Wang,et al.  Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. , 1991, Gene.

[66]  A. Brown,et al.  The antibody response to salmonellae in mice and humans studied by immunoblots and ELISA. , 1989, Microbial pathogenesis.

[67]  E. Bambirra,et al.  Intragastric infection of germfree and conventional mice with Salmonella typhimurium. , 1989, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[68]  M. Blaser,et al.  A review of human salmonellosis: II. Duration of excretion following infection with nontyphi Salmonella. , 1984, Reviews of infectious diseases.

[69]  B. Stocker,et al.  Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines , 1981, Nature.

[70]  D. van der Waaij,et al.  Colonization resistance of the digestive tract in conventional and antibiotic-treated mice. , 1971 .

[71]  D. van der Waaij,et al.  Colonization resistance of the digestive tract in conventional and antibiotic-treated mice , 1971, Journal of Hygiene.

[72]  M. Bohnhoff,et al.  Effect of Streptomycin on Susceptibility of Intestinal Tract to Experimental Salmonella Infection.∗ , 1954, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.