In vivo and in vitro adjuvant activities of the B subunit of Type IIb heat-labile enterotoxin (LT-IIb-B5) from Escherichia coli.

The pentameric B subunit of the Escherichia coli LT-IIb enterotoxin (LT-IIb-B(5)) activates TLR2 signaling in macrophages. Herein we demonstrate that LT-IIb-B(5), in contrast to a TLR2-nonbinding point mutant, induces functional activation of bone marrow-derived dendritic cells and stimulates CD4(+) T cell proliferation, activities which suggested that LT-IIb-B(5) might function as an adjuvant in vivo. Indeed, in an intranasal mouse immunization model, LT-IIb-B(5) augmented specific mucosal and serum antibody responses to a co-administered immunogen, at levels which were almost comparable to those induced by intact LT-IIb holotoxin, a potent but toxic adjuvant. Therefore, LT-IIb-B(5) displays useful adjuvant properties which, combined with lack of enterotoxicity and relative stability against degradation, may find application in mucosal vaccines.

[1]  Shizuo Akira,et al.  Toll-like receptor signalling , 2004, Nature Reviews Immunology.

[2]  R. Tapping,et al.  Mapping of a Microbial Protein Domain Involved in Binding and Activation of the TLR2/TLR1 Heterodimer1 , 2009, The Journal of Immunology.

[3]  J. Clements,et al.  Mucosal adjuvants. , 2005, Vaccine.

[4]  W. Hol,et al.  Crystal structure of a new heat-labile enterotoxin, LT-IIb. , 1996, Structure.

[5]  G. Dougan,et al.  Structure and mucosal adjuvanticity of cholera and Escherichia coli heat-labile enterotoxins. , 1999, Immunology today.

[6]  G. Schuler,et al.  An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. , 1999, Journal of immunological methods.

[7]  T. D. Connell,et al.  Cholera toxin, LT-I, LT-IIa and LT-IIb: the critical role of ganglioside binding in immunomodulation by Type I and Type II heat-labile enterotoxins , 2007, Expert review of vaccines.

[8]  O. Leavy,et al.  Effects of cholera toxin on innate and adaptive immunity and its application as an immunomodulatory agent , 2004, Journal of leukocyte biology.

[9]  P. Godowski,et al.  Microbial Lipopeptides Stimulate Dendritic Cell Maturation Via Toll-Like Receptor 21 , 2001, The Journal of Immunology.

[10]  R. Rappuoli,et al.  Mutants of the Escherichia coli heat-labile enterotoxin as safe and strong adjuvants for intranasal delivery of vaccines , 2003, Expert review of vaccines.

[11]  C. Czerkinsky,et al.  Mucosal immunisation and adjuvants: a brief overview of recent advances and challenges. , 2003, Vaccine.

[12]  B. Pulendran,et al.  Yeast zymosan, a stimulus for TLR2 and dectin-1, induces regulatory antigen-presenting cells and immunological tolerance. , 2006, The Journal of clinical investigation.

[13]  G. Hajishengallis,et al.  Lipid raft‐dependent uptake, signalling and intracellular fate of Porphyromonas gingivalis in mouse macrophages , 2008, Cellular microbiology.

[14]  T. D. Connell,et al.  In Vitro Induction of Immunoglobulin A (IgA)- and IgM-Secreting Plasma Blasts by Cholera Toxin Depends on T-Cell Help and Is Mediated by CD154 Up-Regulation and Inhibition of Gamma Interferon Synthesis , 2007, Infection and Immunity.

[15]  B. Pulendran,et al.  The science of adjuvants , 2007, Expert review of vaccines.

[16]  R. Steinman,et al.  Dendritic cells and the control of immunity , 1998, Nature.

[17]  Karla J. F. Satchell Activation and suppression of the proinflammatory immune response by Vibrio cholerae toxins. , 2003, Microbes and infection.

[18]  R. Tapping,et al.  Ganglioside GD1a Is an Essential Coreceptor for Toll-like Receptor 2 Signaling in Response to the B subunit of Type IIb Enterotoxin* , 2007, Journal of Biological Chemistry.

[19]  W. Hol,et al.  Crystal structure of a new heat-labile enterotoxin, LT-11b, that can ADP-ribosylate Gs-alpha , 1996 .

[20]  C. Sousa,et al.  Inflammatory mediators are insufficient for full dendritic cell activation and promote expansion of CD4+ T cell populations lacking helper function , 2005, Nature Immunology.

[21]  A. Bowie,et al.  The family of five: TIR-domain-containing adaptors in Toll-like receptor signalling , 2007, Nature Reviews Immunology.

[22]  F. Yoshimura,et al.  Differential interactions of fimbriae and lipopolysaccharide from Porphyromonas gingivalis with the Toll‐like receptor 2‐centred pattern recognition apparatus , 2006, Cellular microbiology.

[23]  A. Iwasaki,et al.  Toll-like receptor control of the adaptive immune responses , 2004, Nature Immunology.

[24]  R. Tapping,et al.  The Type II Heat-Labile Enterotoxins LT-IIa and LT-IIb and Their Respective B Pentamers Differentially Induce and Regulate Cytokine Production in Human Monocytic Cells , 2004, Infection and Immunity.

[25]  O. Leavy,et al.  Cholera Toxin Promotes the Induction of Regulatory T Cells Specific for Bystander Antigens by Modulating Dendritic Cell Activation 1 , 2003, The Journal of Immunology.

[26]  T. D. Connell,et al.  Mucosal Adjuvant Properties of Mutant LT-IIa and LT-IIb Enterotoxins That Exhibit Altered Ganglioside-Binding Activities , 2005, Infection and Immunity.

[27]  T. D. Connell,et al.  The A Subunit of Type IIb Enterotoxin (LT-IIb) Suppresses the Proinflammatory Potential of the B Subunit and Its Ability to Recruit and Interact with TLR21 , 2007, The Journal of Immunology.

[28]  R. Steinman,et al.  The Linkage of Innate to Adaptive Immunity via Maturing Dendritic Cells In Vivo Requires CD40 Ligation in Addition to Antigen Presentation and CD80/86 Costimulation , 2004, The Journal of experimental medicine.

[29]  R. Rappuoli,et al.  Modulation of Innate and Acquired Immune Responses by Escherichia coli Heat-Labile Toxin: Distinct Pro- and Anti-Inflammatory Effects of the Nontoxic AB Complex and the Enzyme Activity1 , 2000, The Journal of Immunology.

[30]  M. Russell,et al.  Affinity and Specificity of the Interactions between Streptococcus mutans Antigen I/II and Salivary Components , 1994, Journal of dental research.

[31]  R. Tapping,et al.  Toll-Like Receptor 2 Mediates Cellular Activation by the B Subunits of Type II Heat-Labile Enterotoxins , 2005, Infection and Immunity.

[32]  T. D. Connell,et al.  Immunomodulation with Enterotoxins for the Generation of Secretory Immunity or Tolerance: Applications for Oral Infections , 2005, Journal of dental research.

[33]  R. Schwartz,et al.  Molecular regulation of interleukin‐2 expression by CD28 co‐stimulation and anergy , 1998, Immunological reviews.

[34]  C. Giampaolo,et al.  Comparative study of Clostridium difficile toxin A and cholera toxin in rabbit ileum. , 1989, Gastroenterology.

[35]  R. Steinman,et al.  The Induction of Tolerance by Dendritic Cells That Have Captured Apoptotic Cells , 2000, The Journal of experimental medicine.

[36]  Shizuo Akira,et al.  Toll-like receptors as adjuvant receptors. , 2002, Biochimica et biophysica acta.

[37]  S. Dimmeler,et al.  The Role of Toll-like Receptors (TLRs) in Bacteria-induced Maturation of Murine Dendritic Cells (DCs) , 2001, The Journal of Biological Chemistry.

[38]  G. Dougan,et al.  Immunomodulation Using Bacterial Enterotoxins , 2001, Scandinavian journal of immunology.

[39]  S. Michalek,et al.  Comparative Analysis of the Mucosal Adjuvanticity of the Type II Heat-Labile Enterotoxins LT-IIa and LT-IIb , 2000, Infection and Immunity.

[40]  S. Lehar,et al.  The CD28-related molecule ICOS is required for effective T cell-dependent immune responses. , 2000, Immunity.

[41]  R. Finkelstein,et al.  Subunit number and arrangement in Escherichia coli heat-labile enterotoxin , 1981, Infection and immunity.

[42]  M. Russell,et al.  Mucosal immunization with a bacterial protein antigen genetically coupled to cholera toxin A2/B subunits. , 1995, Journal of immunology.

[43]  T. Lehner,et al.  Protein antigens of Streptococcus mutans: purification and properties of a double antigen and its protease-resistant component , 1980, Infection and immunity.