Hsp65-Producing Lactococcus lactis Prevents Inflammatory Intestinal Disease in Mice by IL-10- and TLR2-Dependent Pathways

Heat shock proteins (Hsps) are highly expressed at all sites of inflammation. As they are ubiquitous and immunodominant antigens, these molecules represent good candidates for the therapeutic use of oral tolerance in autoimmune and chronic inflammatory diseases. Evidences from human and animal studies indicate that inflammatory bowel disease (IBD) results from uncontrolled inflammatory responses to intestinal microbiota. Hsps are immunodominant proteins expressed by several immune cells and by commensal bacteria. Using an IBD mouse model, we showed that oral pretreatment with genetically modified Lactococcus lactis that produces and releases Mycobacterium Hsp65, completely prevented DSS-induced colitis in C57BL/6 mice. Protection was associated with reduced pro-inflammatory cytokines, such as IFN-γ, IL-6, and TNF-α; increased IL-10 production in colonic tissue; and expansion of CD4+Foxp3+ and CD4+LAP+ regulatory T cells in spleen and mesenteric lymph nodes. This effect was dependent on IL-10 and toll-like receptor 2. Thus, this approach may open alternative options for long-term management of IBD.

[1]  A. C. Gomes-Santos,et al.  Antigen administration by continuous feeding enhances oral tolerance and leads to long-lasting effects. , 2015, Journal of immunological methods.

[2]  G. Fitzgerald,et al.  From field to fermentation: the origins of Lactococcus lactis and its domestication to the dairy environment. , 2015, Food microbiology.

[3]  J. Fichna,et al.  Current overview of extrinsic and intrinsic factors in etiology and progression of inflammatory bowel diseases , 2014, Pharmacological reports : PR.

[4]  Martyn F. Symmons,et al.  Assembly and localization of Toll-like receptor signalling complexes , 2014, Nature Reviews Immunology.

[5]  S. Targan,et al.  Microbiota activation and regulation of innate and adaptive immunity , 2014, Immunological reviews.

[6]  H. Al‐Salami,et al.  Inflammatory bowel disease: clinical aspects and treatments , 2014, Journal of inflammation research.

[7]  C. Huttenhower,et al.  Inflammatory bowel disease as a model for translating the microbiome. , 2014, Immunity.

[8]  A. Miyoshi,et al.  Oral Combined Therapy with Probiotics and Alloantigen Induces B Cell–Dependent Long-Lasting Specific Tolerance , 2014, The Journal of Immunology.

[9]  Kunhua Wang,et al.  Heat Shock Factor 2 Levels Are Associated with the Severity of Ulcerative Colitis , 2014, PloS one.

[10]  C. Bonorino,et al.  Extracellular Hsp70 inhibits pro-inflammatory cytokine production by IL-10 driven down-regulation of C/EBPβ and C/EBPδ , 2013, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[11]  B. Corthésy,et al.  Multi-Faceted Functions of Secretory IgA at Mucosal Surfaces , 2013, Front. Immunol..

[12]  D. V. van Bockel,et al.  Innate and Adaptive Immunity in Long-Term Non-Progression in HIV Disease , 2013, Front. Immunol..

[13]  H. Weiner,et al.  Hsp65-producing Lactococcus lactis prevents experimental autoimmune encephalomyelitis in mice by inducing CD4+LAP+ regulatory T cells. , 2013, Journal of autoimmunity.

[14]  N. Vaz,et al.  Oral tolerance correlates with high levels of lymphocyte activity. , 2012, Cellular immunology.

[15]  C. Bonorino,et al.  The anti-inflammatory mechanisms of Hsp70 , 2012, Front. Immun..

[16]  A. Miyoshi,et al.  Cytoplasmic and extracellular expression of pharmaceutical-grade mycobacterial 65-kDa heat shock protein in Lactococcus lactis. , 2012, Genetics and molecular research : GMR.

[17]  J. Lafaille,et al.  Induced CD4+Foxp3+ regulatory T cells in immune tolerance. , 2012, Annual review of immunology.

[18]  A. Mowat,et al.  Oral tolerance to food protein , 2012, Mucosal Immunology.

[19]  D. C. Cara,et al.  New Insights into the Immunological Changes in IL-10-Deficient Mice during the Course of Spontaneous Inflammation in the Gut Mucosa , 2012, Clinical & developmental immunology.

[20]  V. Coelho,et al.  HSP60: Issues and Insights on Its Therapeutic Use as an Immunoregulatory Agent , 2012, Front. Immun..

[21]  A. Miyoshi,et al.  Lactococcus lactis as a live vector: heterologous protein production and DNA delivery systems. , 2011, Protein expression and purification.

[22]  G. Panayi,et al.  Resolution‐associated molecular patterns (RAMP): RAMParts defending immunological homeostasis? , 2011, Clinical and experimental immunology.

[23]  Fiona Powrie,et al.  Intestinal homeostasis and its breakdown in inflammatory bowel disease , 2011, Nature.

[24]  H. Weiner,et al.  Oral tolerance , 2011, Immunological reviews.

[25]  S. Albani,et al.  Heat Shock Protein-Derived T-Cell Epitopes Contribute to Autoimmune Inflammation in Pediatric Crohn's Disease , 2009, PloS one.

[26]  Mahavir Singh,et al.  IL-10 Is Critically Involved in Mycobacterial HSP70 Induced Suppression of Proteoglycan-Induced Arthritis , 2009, PloS one.

[27]  E. Chang,et al.  Do heat shock proteins play any role in gut inflammation? , 2008, Inflammatory bowel diseases.

[28]  A. Sartori,et al.  Immunization with pVAXhsp65 Decreases Inflammation and Modulates Immune Response in Experimental Encephalomyelitis , 2008, Neuroimmunomodulation.

[29]  Makiya Nishikawa,et al.  Heat shock protein derivatives for delivery of antigens to antigen presenting cells. , 2008, International journal of pharmaceutics.

[30]  I. Cohen,et al.  Stress, Heat Shock Proteins, and Autoimmunity , 2007, Annals of the New York Academy of Sciences.

[31]  M. Chamaillard,et al.  Prevention and treatment of colitis with Lactococcus lactis secreting the immunomodulatory Yersinia LcrV protein. , 2007, Gastroenterology.

[32]  C. Bonorino,et al.  Mycobacterium tuberculosis heat‐shock protein 70 impairs maturation of dendritic cells from bone marrow precursors, induces interleukin‐10 production and inhibits T‐cell proliferation in vitro , 2007, Immunology.

[33]  G Gerken,et al.  Toll-like receptor 2 controls mucosal inflammation by regulating epithelial barrier function. , 2007, Gastroenterology.

[34]  Ruslan Medzhitov,et al.  Role of toll-like receptors in spontaneous commensal-dependent colitis. , 2006, Immunity.

[35]  H. Weiner,et al.  Oral Tolerance: Therapeutic Implications for Autoimmune Diseases , 2006, Clinical & developmental immunology.

[36]  F. Powrie,et al.  Regulatory T cells and intestinal homeostasis , 2005, Immunological reviews.

[37]  B. Prakken,et al.  Heat-shock proteins induce T-cell regulation of chronic inflammation , 2005, Nature Reviews Immunology.

[38]  C. Ramos,et al.  Protein folding assisted by chaperones. , 2005, Protein and peptide letters.

[39]  M. Tsan,et al.  Endogenous ligands of Toll‐like receptors , 2004, Journal of leukocyte biology.

[40]  W. Kuis,et al.  CD4+CD25bright Regulatory T Cells Actively Regulate Inflammation in the Joints of Patients with the Remitting Form of Juvenile Idiopathic Arthritis , 2004, The Journal of Immunology.

[41]  A. Zuercher,et al.  Interactions of commensal gut microbes with subsets of B- and T-cells in the murine host. , 2004, Vaccine.

[42]  P. Srivastava,et al.  Heat‐Shock Proteins , 2003, Current protocols in immunology.

[43]  I. Cohen,et al.  T cells respond to heat shock protein 60 via TLR2: activation of adhesion and inhibition of chemokine receptors , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[44]  Toru Takahashi,et al.  Direct Binding of Toll-Like Receptor 2 to Zymosan, and Zymosan-Induced NF-κB Activation and TNF-α Secretion Are Down-Regulated by Lung Collectin Surfactant Protein A1 , 2003, The Journal of Immunology.

[45]  H. Weiner,et al.  Oral tolerance induced by continuous feeding: enhanced up-regulation of transforming growth factor-beta/interleukin-10 and suppression of experimental autoimmune encephalomyelitis. , 2003, Journal of autoimmunity.

[46]  P. Libby,et al.  Mucosal Administration of Heat Shock Protein-65 Decreases Atherosclerosis and Inflammation in Aortic Arch of Low-Density Lipoprotein Receptor-Deficient Mice , 2002, Circulation.

[47]  Johannes Buchner,et al.  Molecular chaperones--cellular machines for protein folding. , 2002, Angewandte Chemie.

[48]  K. Ramaswamy,et al.  Colon epithelial cellular protein induces oral tolerance in the experimental model of colitis by trinitrobenzene sulfonic acid. , 2001, The Journal of laboratory and clinical medicine.

[49]  W. Fiers,et al.  Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. , 2000, Science.

[50]  K. Seno,et al.  Involvement of Luminal Bacteria, Heat Shock Protein 60, Macrophages and γδ T Cells in Dextran Sulfate Sodium-Induced Colitis in Rats , 2000, Digestive Diseases and Sciences.

[51]  J. Wallace,et al.  Spontaneously developing chronic colitis in IL-10/iNOS double-deficient mice. , 2000, American journal of physiology. Gastrointestinal and liver physiology.

[52]  I. Cohen,et al.  The pathogenicity of islet‐infiltrating lymphocytes in the non‐obese diabetic (NOD) mouse , 1999, Clinical and experimental immunology.

[53]  B. Stransky,et al.  Aging affects oral tolerance induction but not its maintenance in mice , 1998, Mechanisms of Ageing and Development.

[54]  T. Macdonald.  Breakdown of tolerance to the intestinal bacterial flora in inflammatory bowel disease (IBD) , 1995, Clinical and experimental immunology.

[55]  K. Rajewsky,et al.  Interleukin-10-deficient mice develop chronic enterocolitis , 1993, Cell.

[56]  H. Cooper,et al.  Treatment of dextran sulfate sodium-induced murine colitis by intracolonic cyclosporin , 1993, Digestive Diseases and Sciences.

[57]  D. Rampton,et al.  Circulating antibodies to heat‐shock protein 60 in Crohn's disease and ulcerative colitis , 1992, Clinical and experimental immunology.

[58]  I. Cohen,et al.  Autoimmunity, microbial immunity and the immunological homunculus. , 1991, Immunology today.

[59]  D. Parrott,et al.  Preparation and purification of lymphocytes from the epithelium and lamina propria of murine small intestine. , 1981, Gut.

[60]  I. Cohen,et al.  Heat shock protein 60 enhances CD4+CD25+ regulatory T cell function via innate TLR2 signaling. , 2018, The Journal of clinical investigation.

[61]  P. van Baarlen,et al.  Host-recognition of pathogens and commensals in the mammalian intestine. , 2013, Current topics in microbiology and immunology.

[62]  E. Szigethy,et al.  Inflammatory bowel disease. , 2011, Pediatric clinics of North America.

[63]  E. Remaut,et al.  Orally administered L. lactis secreting an anti-TNF Nanobody demonstrate efficacy in chronic colitis , 2010, Mucosal Immunology.

[64]  Toru Takahashi,et al.  Direct binding of Toll-like receptor 2 to zymosan, and zymosan-induced NF-kappa B activation and TNF-alpha secretion are down-regulated by lung collectin surfactant protein A. , 2003, Journal of immunology.

[65]  K. Seno,et al.  Involvement of luminal bacteria, heat shock protein 60, macrophages and gammadelta T cells in dextran sulfate sodium-induced colitis in rats. , 2000, Digestive diseases and sciences.

[66]  H. Weiner,et al.  Oral tolerance: mechanisms and therapeutic applications. , 1999, Advances in immunology.

[67]  R. Sartor.  Pathogenesis and immune mechanisms of chronic inflammatory bowel diseases. , 1997, The American journal of gastroenterology.