Uptake through glycoprotein 2 of FimH+ bacteria by M cells initiates mucosal immune response

The mucosal immune system forms the largest part of the entire immune system, containing about three-quarters of all lymphocytes and producing grams of secretory IgA daily to protect the mucosal surface from pathogens. To evoke the mucosal immune response, antigens on the mucosal surface must be transported across the epithelial barrier into organized lymphoid structures such as Peyer’s patches. This function, called antigen transcytosis, is mediated by specialized epithelial M cells. The molecular mechanisms promoting this antigen uptake, however, are largely unknown. Here we report that glycoprotein 2 (GP2), specifically expressed on the apical plasma membrane of M cells among enterocytes, serves as a transcytotic receptor for mucosal antigens. Recombinant GP2 protein selectively bound a subset of commensal and pathogenic enterobacteria, including Escherichia coli and Salmonella enterica serovar Typhimurium (S. Typhimurium), by recognizing FimH, a component of type I pili on the bacterial outer membrane. Consistently, these bacteria were colocalized with endogenous GP2 on the apical plasma membrane as well as in cytoplasmic vesicles in M cells. Moreover, deficiency of bacterial FimH or host GP2 led to defects in transcytosis of type-I-piliated bacteria through M cells, resulting in an attenuation of antigen-specific immune responses in Peyer’s patches. GP2 is therefore a previously unrecognized transcytotic receptor on M cells for type-I-piliated bacteria and is a prerequisite for the mucosal immune response to these bacteria. Given that M cells are considered a promising target for oral vaccination against various infectious diseases, the GP2-dependent transcytotic pathway could provide a new target for the development of M-cell-targeted mucosal vaccines.

[1]  N. Mantis,et al.  Collaboration of epithelial cells with organized mucosal lymphoid tissues , 2001, Nature Immunology.

[2]  H. Kiyono,et al.  Comprehensive Gene Expression Profiling of Peyer’s Patch M Cells, Villous M-Like Cells, and Intestinal Epithelial Cells1 , 2008, The Journal of Immunology.

[3]  V. Sukhatme,et al.  GP-2/THP gene family encodes self-binding glycosylphosphatidylinositol-anchored proteins in apical secretory compartments of pancreas and kidney , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Keiichiro Suzuki,et al.  Critical Roles of Activation-Induced Cytidine Deaminase in the Homeostasis of Gut Flora , 2002, Science.

[5]  J. Glasner,et al.  Gene replacement without selection: regulated suppression of amber mutations in Escherichia coli. , 2003, Gene.

[6]  M. Rescigno,et al.  The biology of intestinal immunoglobulin A responses. , 2008, Immunity.

[7]  T. Hoops,et al.  Isolation of the cDNA encoding glycoprotein-2 (GP-2), the major zymogen granule membrane protein. Homology to uromodulin/Tamm-Horsfall protein. , 1991, The Journal of biological chemistry.

[8]  J. Sirard,et al.  Live attenuated Salmonella: a paradigm of mucosal vaccines , 1999, Immunological reviews.

[9]  T. Hoops,et al.  The pancreatic membrane protein GP-2 localizes specifically to secretory granules and is shed into the pancreatic juice as a protein aggregate. , 1990, European journal of cell biology.

[10]  F. Collins,et al.  Experimental Yersinia enterocolitica infection in mice: Kinetics of growth , 1974, Infection and immunity.

[11]  B. Hirst,et al.  M-Cell Surface β1 Integrin Expression and Invasin-Mediated Targeting of Yersinia pseudotuberculosis to Mouse Peyer’s Patch M Cells , 1998, Infection and Immunity.

[12]  S. Fukuyama,et al.  A novel M cell–specific carbohydrate-targeted mucosal vaccine effectively induces antigen-specific immune responses , 2007, The Journal of experimental medicine.

[13]  J. Kraehenbuhl,et al.  A recombinant Salmonella typhimurium vaccine strain is taken up and survives within murine Peyer's patch dendritic cells , 2000, Cellular microbiology.

[14]  J. Kraehenbuhl,et al.  Epithelial M cells: differentiation and function. , 2000, Annual review of cell and developmental biology.

[15]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Mcghee,et al.  Optimizing oral vaccines: induction of systemic and mucosal B-cell and antibody responses to tetanus toxoid by use of cholera toxin as an adjuvant , 1993, Infection and immunity.

[17]  J. Mcghee,et al.  Alternate Mucosal Immune System: Organized Peyer’s Patches Are Not Required for IgA Responses in the Gastrointestinal Tract1 , 2000, The Journal of Immunology.

[18]  S. Fukuoka,et al.  GP2/THP gene family of self-binding, GPI-anchored proteins forms a cluster at chromosome 7F1 region in mouse genome. , 2004, Biochemical and biophysical research communications.

[19]  Hong-ying Huang,et al.  Ablation of the Tamm-Horsfall protein gene increases susceptibility of mice to bladder colonization by type 1-fimbriated Escherichia coli. , 2004, American journal of physiology. Renal physiology.

[20]  J. Gordon,et al.  IgA response to symbiotic bacteria as a mediator of gut homeostasis. , 2007, Cell host & microbe.

[21]  P. Kujala,et al.  Clusterin in human gut-associated lymphoid tissue, tonsils, and adenoids: localization to M cells and follicular dendritic cells , 2008, Histochemistry and Cell Biology.

[22]  D. McDonald,et al.  Recruitment of HIV and Its Receptors to Dendritic Cell-T Cell Junctions , 2003, Science.

[23]  E. Allen-Vercoe,et al.  Salmonella enterica var Typhimurium and Salmonella enterica var Enteritidis express type 1 fimbriae in the rat in vivo. , 1997, FEMS immunology and medical microbiology.

[24]  J. Hughes,et al.  Analysis of Host Cells Associated with the Spv-Mediated Increased Intracellular Growth Rate of Salmonella typhimurium in Mice , 1998, Infection and Immunity.

[25]  S. Craig,et al.  PEYER'S PATCHES: AN ENRICHED SOURCE OF PRECURSORS FOR IGA-PRODUCING IMMUNOCYTES IN THE RABBIT , 1971, The Journal of experimental medicine.

[26]  H. Kiyono,et al.  Peyer's Patches Are Required for Intestinal Immunoglobulin A Responses to Salmonella spp , 2007, Infection and Immunity.

[27]  M. Komatsu,et al.  Biochemical and morphological detection of inclusion bodies in autophagy-deficient mice. , 2009, Methods in enzymology.

[28]  J. Mcghee,et al.  Regulation of mucosal and systemic antibody responses by T helper cell subsets, macrophages, and derived cytokines following oral immunization with live recombinant Salmonella. , 1996, Journal of immunology.

[29]  T. Sasazuki,et al.  T helper type 2 differentiation and intracellular trafficking of the interleukin 4 receptor-α subunit controlled by the Rac activator Dock2 , 2007, Nature Immunology.

[30]  P. Sansonetti,et al.  M cells as ports of entry for enteroinvasive pathogens: mechanisms of interaction, consequences for the disease process. , 1999, Seminars in immunology.

[31]  M. Loeffelholz,et al.  PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid , 1994, Journal of clinical microbiology.

[32]  K. Kawano,et al.  Distinct gene expression profiles characterize cellular phenotypes of follicle-associated epithelium and M cells. , 2005, DNA research : an international journal for rapid publication of reports on genes and genomes.

[33]  A. Iwasaki,et al.  Localization of Distinct Peyer's Patch Dendritic Cell Subsets and Their Recruitment by Chemokines Macrophage Inflammatory Protein (Mip)-3α, Mip-3β, and Secondary Lymphoid Organ Chemokine , 2000, The Journal of experimental medicine.

[34]  A. Lowe,et al.  The pancreatic zymogen granule membrane protein, GP2, binds Escherichia coli type 1 Fimbriae , 2009, BMC gastroenterology.

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

[36]  A. Jones,et al.  Epithelial cell specialization within human Peyer's patches: an ultrastructural study of intestinal lymphoid follicles. , 1974, Gastroenterology.

[37]  S. Yonehara,et al.  The Membrane-Bound Chemokine CXCL16 Expressed on Follicle-Associated Epithelium and M Cells Mediates Lympho-Epithelial Interaction in GALT1 , 2006, The Journal of Immunology.

[38]  P. Cossart,et al.  Bacterial Adhesion and Entry into Host Cells , 2006, Cell.

[39]  S. Michie,et al.  Absence of the Major Zymogen Granule Membrane Protein, GP2, Does Not Affect Pancreatic Morphology or Secretion* , 2004, Journal of Biological Chemistry.