Diverse Intestinal Bacteria Contain Putative Zwitterionic Capsular Polysaccharides with Anti-inflammatory Properties.

Zwitterionic capsular polysaccharides (ZPSs) are bacterial products that modulate T cells, including inducing anti-inflammatory IL-10-secreting T regulatory cells (Tregs). However, only a few diverse bacteria are known to modulate the host immune system via ZPS. We present a genomic screen for bacteria encoding ZPS molecules. We identify diverse host-associated bacteria, including commensals and pathogens with known anti-inflammatory properties, with the capacity to produce ZPSs. Human mononuclear cells stimulated with lysates from putative ZPS-producing bacteria induce significantly greater IL-10 production and higher proportions of Tregs than lysates from non-ZPS-encoding relatives or a commensal strain of Bacteroides cellulosilyticus in which a putative ZPS biosynthetic operon was genetically disrupted. Similarly, wild-type B. cellulosilyticus DSM 14838, but not a close relative lacking a putative ZPS, attenuated experimental colitis in mice. Collectively, this screen identifies bacterial strains that may use ZPSs to interact with the host as well as those with potential probiotic properties.

[1]  T. Chatila,et al.  The Toll-Like Receptor 2 Pathway Establishes Colonization by a Commensal of the Human Microbiota , 2011, Science.

[2]  B. Palmer,et al.  Impaired Function of CTLA-4 in the Lungs of Patients with Chronic Beryllium Disease Contributes to Persistent Inflammation , 2013, The Journal of Immunology.

[3]  Kunyan Zhang,et al.  Bacteremia Caused by Clostridium symbiosum , 2004, Journal of Clinical Microbiology.

[4]  Frank Oliver Glöckner,et al.  Graphical representation of ribosomal RNA probe accessibility data using ARB software package , 2005, BMC Bioinformatics.

[5]  A. Rudensky,et al.  Single-cell analysis of normal and FOXP3-mutant human T cells: FOXP3 expression without regulatory T cell development. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[6]  T. Hudcovic,et al.  Oral administration of Parabacteroides distasonis antigens attenuates experimental murine colitis through modulation of immunity and microbiota composition , 2011, Clinical and experimental immunology.

[7]  B. Polk,et al.  Bacteroides fragilis subspecies in clinical isolates. , 1977, Annals of internal medicine.

[8]  A. Weintraub,et al.  Virulence factors in infections with bacteroides fragilis: isolation and characterization of capsular polysaccharide and lipopolysaccharide. , 1982, Scandinavian journal of infectious diseases. Supplementum.

[9]  Ying Xu,et al.  DOOR: a database for prokaryotic operons , 2008, Nucleic Acids Res..

[10]  P. Lawson,et al.  Bacteroides cellulosilyticus sp. nov., a cellulolytic bacterium from the human gut microbial community. , 2007, International journal of systematic and evolutionary microbiology.

[11]  J. Bartlett,et al.  The capsular polysaccharide of Bacteroides fragilis as a virulence factor: comparison of the pathogenic potential of encapsulated and unencapsulated strains. , 1977, The Journal of infectious diseases.

[12]  Y. Belkaid,et al.  Intestinal microbiota: shaping local and systemic immune responses. , 2012, Seminars in immunology.

[13]  D. Kasper,et al.  The yin yang of bacterial polysaccharides: lessons learned from B. fragilis PSA , 2012, Immunological reviews.

[14]  C. Dinarello,et al.  Alpha-1-antitrypsin Therapy Ameliorates Acute Colitis and Chronic Murine Ileitis , 2013, Inflammatory bowel diseases.

[15]  J. Richards,et al.  Structural characterization and MHCII-dependent immunological properties of the zwitterionic O-chain antigen of Morganella morganii. , 2011, Glycobiology.

[16]  Klaus Ley,et al.  Bacterial colonization factors control specificity and stability of the gut microbiota , 2013, Nature.

[17]  C. Weidenmaier,et al.  The Zwitterionic Cell Wall Teichoic Acid of Staphylococcus aureus Provokes Skin Abscesses in Mice by a Novel CD4+ T-Cell-Dependent Mechanism , 2010, PloS one.

[18]  H. Kiyono,et al.  Probiotic Bifidobacterium breve Induces IL-10-Producing Tr1 Cells in the Colon , 2012, PLoS pathogens.

[19]  D. Kasper,et al.  A commensal symbiotic factor derived from Bacteroides fragilis promotes human CD39+Foxp3+ T cells and Treg function , 2015, Gut microbes.

[20]  Robert C. Edgar,et al.  MUSCLE: multiple sequence alignment with high accuracy and high throughput. , 2004, Nucleic acids research.

[21]  A. Macpherson,et al.  Immune adaptations that maintain homeostasis with the intestinal microbiota , 2010, Nature Reviews Immunology.

[22]  S. Mazmanian,et al.  Outer membrane vesicles of a human commensal mediate immune regulation and disease protection. , 2012, Cell host & microbe.

[23]  Katherine O'Riordan,et al.  Staphylococcus aureus Capsular Polysaccharides , 2004, Clinical Microbiology Reviews.

[24]  M. Hattori,et al.  Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota , 2013, Nature.

[25]  S. Robson,et al.  An intestinal commensal symbiosis factor controls neuroinflammation via TLR2-mediated CD39 signaling , 2014, Nature Communications.

[26]  Y. Belkaid.  Regulatory T cells and infection: a dangerous necessity , 2007, Nature Reviews Immunology.

[27]  S. Mazmanian,et al.  Inducible Foxp3+ regulatory T-cell development by a commensal bacterium of the intestinal microbiota , 2010, Proceedings of the National Academy of Sciences.

[28]  Bernard Henrissat,et al.  Effects of Diet on Resource Utilization by a Model Human Gut Microbiota Containing Bacteroides cellulosilyticus WH2, a Symbiont with an Extensive Glycobiome , 2013, PLoS biology.

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

[30]  Lori S C Kreisman,et al.  Glycoantigens Induce Human Peripheral Tr1 Cell Differentiation with Gut-homing Specialization* , 2011, The Journal of Biological Chemistry.

[31]  T. Vicsek,et al.  Uncovering the overlapping community structure of complex networks in nature and society , 2005, Nature.

[32]  H. Ploegh,et al.  Zwitterionic Polysaccharides Stimulate T Cells by MHC Class II-Dependent Interactions1 , 2002, The Journal of Immunology.

[33]  B. Palmer,et al.  Lymphocytic alveolitis is associated with the accumulation of functionally impaired HIV-specific T cells in the lung of antiretroviral therapy-naive subjects. , 2015, American journal of respiratory and critical care medicine.

[34]  C. Hsieh,et al.  Peripheral education of the immune system by colonic commensal microbiota , 2011, Nature.

[35]  K. Mills,et al.  Regulatory T cells: friend or foe in immunity to infection? , 2004, Nature Reviews Immunology.

[36]  T. Stephen,et al.  The Modulation of Adaptive Immune Responses by Bacterial Zwitterionic Polysaccharides , 2010, International journal of microbiology.

[37]  R. Plattner,et al.  Identification of L-iduronic acid as a constituent of the major extracellular polysaccharide produced by Butyrivibrio fibrisolvens strain X6C61 , 1988 .

[38]  William A. Walters,et al.  QIIME allows analysis of high-throughput community sequencing data , 2010, Nature Methods.

[39]  Lynn K. Carmichael,et al.  A Genomic View of the Human-Bacteroides thetaiotaomicron Symbiosis , 2003, Science.

[40]  B. Palmer,et al.  HIV-induced alteration in gut microbiota , 2014, Gut microbes.

[41]  Robert C. Edgar,et al.  BIOINFORMATICS APPLICATIONS NOTE , 2001 .

[42]  D. Kasper,et al.  Type I Streptococcus pneumoniae carbohydrate utilizes a nitric oxide and MHC II‐dependent pathway for antigen presentation , 2009, Immunology.

[43]  J. Faith,et al.  Extensive personal human gut microbiota culture collections characterized and manipulated in gnotobiotic mice , 2011, Proceedings of the National Academy of Sciences.

[44]  G. Donelli,et al.  Haemagglutination and surface structures in strains of Clostridium spiroforme. , 1989, FEMS microbiology letters.

[45]  D. Kasper,et al.  Polysaccharide Biosynthesis Locus Required for Virulence of Bacteroides fragilis , 2001, Infection and Immunity.

[46]  Xin Chen,et al.  DOOR 2.0: presenting operons and their functions through dynamic and integrated views , 2013, Nucleic Acids Res..

[47]  Christian L. Lauber,et al.  PrimerProspector: de novo design and taxonomic analysis of barcoded polymerase chain reaction primers , 2011, Bioinform..

[48]  S. Mazmanian,et al.  The love–hate relationship between bacterial polysaccharides and the host immune system , 2006, Nature Reviews Immunology.

[49]  J. Dankert,et al.  Gas gangrene in an immunocompromised girl due to a Clostridium ramosum infection. , 1999, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[50]  Y. Wang,et al.  Structural basis of the abscess-modulating polysaccharide A2 from Bacteroides fragilis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Laurie E. Comstock,et al.  Extensive surface diversity of a commensal microorganism by multiple DNA inversions , 2001, Nature.

[52]  M. Alexeyev.  The pKNOCK series of broad-host-range mobilizable suicide vectors for gene knockout and targeted DNA insertion into the chromosome of gram-negative bacteria. , 1999, BioTechniques.

[53]  Wei Hu,et al.  Location, location, location: tissue‐specific regulation of immune responses , 2013, Journal of leukocyte biology.

[54]  S. Mazmanian,et al.  A microbial symbiosis factor prevents intestinal inflammatory disease , 2008, Nature.

[55]  D. Kasper,et al.  Structural features of polysaccharides that induce intra-abdominal abscesses. , 1993, Science.

[56]  B. Palmer,et al.  Regulation of Virus-Specific CD4+ T Cell Function by Multiple Costimulatory Receptors during Chronic HIV Infection , 2010, The Journal of Immunology.

[57]  S. Mazmanian,et al.  An Immunomodulatory Molecule of Symbiotic Bacteria Directs Maturation of the Host Immune System , 2005, Cell.

[58]  D. Hafler,et al.  FOXP3+ regulatory T cells in the human immune system , 2010, Nature Reviews Immunology.

[59]  F. Avci,et al.  How bacterial carbohydrates influence the adaptive immune system. , 2010, Annual review of immunology.

[60]  J. Bassaganya-Riera,et al.  CD4+ T‐cell responses and distribution at the colonic mucosa during Brachyspira hyodysenteriae‐induced colitis in pigs , 2005, Immunology.