Siderophore vaccine conjugates protect against uropathogenic Escherichia coli urinary tract infection

Significance Urinary tract infections (UTIs) are primarily caused by uropathogenic Escherichia coli (UPEC), and 1 in 40 women experience chronic UTIs during their lifetime. The antibiotic courses required to treat infections promote antibiotic resistance, and current vaccine options offer limited protection. We have pioneered a strategy using small iron-chelating compounds called siderophores as vaccine antigens. These siderophores are not produced by commensal bacteria and are required for UTI. The siderophore vaccines reported here are easy to formulate and reduce bacterial burdens in a murine model of UTI. This report highlights the untapped resource of bacteria-specific small molecules as potential vaccine antigens and provides a proof of principle for incorporating these compounds into multicomponent vaccines for the prevention of bacterial infections. Uropathogenic Escherichia coli (UPEC) is the primary cause of uncomplicated urinary tract infections (UTIs). Whereas most infections are isolated cases, 1 in 40 women experience recurrent UTIs. The rise in antibiotic resistance has complicated the management of chronic UTIs and necessitates new preventative strategies. Currently, no UTI vaccines are approved for use in the United States, and the development of a highly effective vaccine remains elusive. Here, we have pursued a strategy for eliciting protective immunity by vaccinating with small molecules required for pathogenesis, rather than proteins or peptides. Small iron-chelating molecules called siderophores were selected as antigens to vaccinate against UTI for this vaccine strategy. These pathogen-associated stealth siderophores evade host immune defenses and enhance bacterial virulence. Previous animal studies revealed that vaccination with siderophore receptor proteins protects against UTI. The poor solubility of these integral outer-membrane proteins in aqueous solutions limits their practical utility. Because their cognate siderophores are water soluble, we hypothesized that these bacterial-derived small molecules are prime vaccine candidates. To test this hypothesis, we immunized mice with siderophores conjugated to an immunogenic carrier protein. The siderophore–protein conjugates elicited an adaptive immune response that targeted bacterial stealth siderophores and protected against UTI. Our study has identified additional antigens suitable for a multicomponent UTI vaccine and highlights the potential use of bacterial-derived small molecules as antigens in vaccine therapies.

[1]  Yan He,et al.  Siderophore Biosynthesis Governs the Virulence of Uropathogenic Escherichia coli by Coordinately Modulating the Differential Metabolism. , 2016, Journal of proteome research.

[2]  Eric P. Skaar,et al.  Metals in infectious diseases and nutritional immunity. , 2015, Metallomics : integrated biometal science.

[3]  S. Sheppard,et al.  Variation in Siderophore Biosynthetic Gene Distribution and Production across Environmental and Faecal Populations of Escherichia coli , 2015, PloS one.

[4]  D. Rasko,et al.  Host-specific induction of Escherichia coli fitness genes during human urinary tract infection , 2014, Proceedings of the National Academy of Sciences.

[5]  C. Dozois,et al.  Iron, copper, zinc, and manganese transport and regulation in pathogenic Enterobacteria: correlations between strains, site of infection and the relative importance of the different metal transport systems for virulence , 2013, Front. Cell. Infect. Microbiol..

[6]  D. Rasko,et al.  Draft genome sequences of five recent human uropathogenic Escherichia coli isolates. , 2013, Pathogens and disease.

[7]  A. Spivak,et al.  Urinary tract infections: current and emerging management strategies. , 2013, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[8]  H. Mobley,et al.  Immunization with the Yersiniabactin Receptor, FyuA, Protects against Pyelonephritis in a Murine Model of Urinary Tract Infection , 2013, Infection and Immunity.

[9]  Eric P. Skaar,et al.  Iron in infection and immunity. , 2013, Cell host & microbe.

[10]  G. H. Reed,et al.  The ubiquity of iron. , 2012, ACS chemical biology.

[11]  H. Mobley,et al.  Preventing urinary tract infection: progress toward an effective Escherichia coli vaccine , 2012, Expert review of vaccines.

[12]  C. Dozois,et al.  The Ins and Outs of siderophore mediated iron uptake by extra-intestinal pathogenic Escherichia coli. , 2011, Veterinary microbiology.

[13]  T. D. de Reijke,et al.  Cranberries vs antibiotics to prevent urinary tract infections: a randomized double-blind noninferiority trial in premenopausal women. , 2011, Archives of internal medicine.

[14]  C. Dozois,et al.  Klebsiella pneumoniae Yersiniabactin Promotes Respiratory Tract Infection through Evasion of Lipocalin 2 , 2011, Infection and Immunity.

[15]  C. Dozois,et al.  Secretion, but not overall synthesis, of catecholate siderophores contributes to virulence of extraintestinal pathogenic Escherichia coli , 2011, Molecular microbiology.

[16]  C. Alteri,et al.  Identification of In Vivo-Induced Antigens Including an RTX Family Exoprotein Required for Uropathogenic Escherichia coli Virulence , 2011, Infection and Immunity.

[17]  Richard Colgan,et al.  International clinical practice guidelines for the treatment of acute uncomplicated cystitis and pyelonephritis in women: A 2010 update by the Infectious Diseases Society of America and the European Society for Microbiology and Infectious Diseases. , 2011, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[18]  H. Mobley,et al.  Redundancy and Specificity of Escherichia coli Iron Acquisition Systems during Urinary Tract Infection , 2011, Infection and Immunity.

[19]  Betsy Foxman,et al.  The epidemiology of urinary tract infection , 2010, Nature Reviews Urology.

[20]  D. Rasko,et al.  Escherichia coli Global Gene Expression in Urine from Women with Urinary Tract Infection , 2010, PLoS pathogens.

[21]  H. Mobley,et al.  Waging War against Uropathogenic Escherichia coli: Winning Back the Urinary Tract , 2009, Infection and Immunity.

[22]  A. Marchese,et al.  The ARESC study: an international survey on the antimicrobial resistance of pathogens involved in uncomplicated urinary tract infections. , 2009, International journal of antimicrobial agents.

[23]  C. Alteri,et al.  Mucosal Immunization with Iron Receptor Antigens Protects against Urinary Tract Infection , 2009, PLoS pathogens.

[24]  H. Mobley,et al.  Identification of uropathogenic Escherichia coli surface proteins by shotgun proteomics. , 2009, Journal of microbiological methods.

[25]  R. Bergeron,et al.  Vibriobactin antibodies: a vaccine strategy. , 2009, Journal of medicinal chemistry.

[26]  A. Nicolau,et al.  Antibiotherapy and pathogenesis of uncomplicated UTI: difficult relationships , 2009, Journal of applied microbiology.

[27]  C. Alteri,et al.  Fitness of Escherichia coli during Urinary Tract Infection Requires Gluconeogenesis and the TCA Cycle , 2009, PLoS pathogens.

[28]  H. Mobley,et al.  Haem acquisition is facilitated by a novel receptor Hma and required by uropathogenic Escherichia coli for kidney infection , 2009, Molecular microbiology.

[29]  W. Hörl,et al.  Current concepts of molecular defence mechanisms operative during urinary tract infection , 2008, European journal of clinical investigation.

[30]  T. J. Wiles,et al.  Origins and virulence mechanisms of uropathogenic Escherichia coli. , 2008, Experimental and molecular pathology.

[31]  P. Humphrey,et al.  Detection of Intracellular Bacterial Communities in Human Urinary Tract Infection , 2007, PLoS medicine.

[32]  M. Marahiel,et al.  Siderophore-Based Iron Acquisition and Pathogen Control , 2007, Microbiology and Molecular Biology Reviews.

[33]  B. Foxman,et al.  Antibiotic resistance and pyelonephritis. , 2007, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[34]  M. Guzmán,et al.  Kinetics of antibodies in sera, saliva, and urine samples from adult patients with primary or secondary dengue 3 virus infections. , 2007, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[35]  C. Alteri,et al.  Quantitative Profile of the Uropathogenic Escherichia coli Outer Membrane Proteome during Growth in Human Urine , 2007, Infection and Immunity.

[36]  J. Karlowsky,et al.  Fluoroquinolone-Resistant Urinary Isolates of Escherichia coli from Outpatients Are Frequently Multidrug Resistant: Results from the North American Urinary Tract Infection Collaborative Alliance-Quinolone Resistance Study , 2006, Antimicrobial Agents and Chemotherapy.

[37]  L. Nicolle,et al.  Antibiotic resistance in outpatient urinary isolates: final results from the North American Urinary Tract Infection Collaborative Alliance (NAUTICA). , 2005, International journal of antimicrobial agents.

[38]  H. Mobley,et al.  The IrgA Homologue Adhesin Iha Is an Escherichia coli Virulence Factor in Murine Urinary Tract Infection , 2005, Infection and Immunity.

[39]  Shizuo Akira,et al.  Lipocalin 2 mediates an innate immune response to bacterial infection by sequestrating iron , 2004, Nature.

[40]  H. Mobley,et al.  Transcriptome of Uropathogenic Escherichia coli during Urinary Tract Infection , 2004, Infection and Immunity.

[41]  M. Footer,et al.  Differentiation and developmental pathways of uropathogenic Escherichia coli in urinary tract pathogenesis. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[42]  H. Mobley,et al.  Development of an Intranasal Vaccine To Prevent Urinary Tract Infection by Proteus mirabilis , 2004, Infection and Immunity.

[43]  G. Leverson,et al.  Phase 2 clinical trial of a vaginal mucosal vaccine for urinary tract infections. , 2003, The Journal of urology.

[44]  T. Russo,et al.  IroN Functions as a Siderophore Receptor and Is a Urovirulence Factor in an Extraintestinal Pathogenic Isolate of Escherichia coli , 2002, Infection and Immunity.

[45]  R. Strong,et al.  The neutrophil lipocalin NGAL is a bacteriostatic agent that interferes with siderophore-mediated iron acquisition. , 2002, Molecular cell.

[46]  A. Torres,et al.  TonB-Dependent Systems of Uropathogenic Escherichia coli: Aerobactin and Heme Transport and TonB Are Required for Virulence in the Mouse , 2001, Infection and Immunity.

[47]  T. Hooton,et al.  Increasing Antimicrobial Resistance and the Management of Uncomplicated Community-Acquired Urinary Tract Infections , 2001, Annals of Internal Medicine.

[48]  C. Janeway,et al.  Adaptive Immunity to Infection , 2001 .

[49]  D. Le Roy,et al.  Results of passive and active immunization directed against ferric aerobactin in experimental enterobacterial infections in mice and chickens. , 1995, Research in microbiology.

[50]  J. Buyer,et al.  A Method for Detection of Pseudobactin, the Siderophore Produced by a Plant-Growth-Promoting Pseudomonas Strain, in the Barley Rhizosphere , 1993, Applied and environmental microbiology.

[51]  D. Le Roy,et al.  Comparison of chemical assay, bioassay, enzyme-linked immunosorbent assay, and dot blot hybridization for detection of aerobactin in members of the family Enterobacteriaceae , 1993, Applied and environmental microbiology.

[52]  D. Le Roy,et al.  Activity and specificity of a mouse monoclonal antibody to ferric aerobactin , 1992, Infection and immunity.

[53]  E. Graves National Hospital Discharge Survey. , 1989, Vital and health statistics. Series 13, Data from the National Health Survey.

[54]  Urinary albumin, transferrin and iron excretion in diabetic patients. , 1991, Kidney international.

[55]  H. Tammen Immunobiotherapy with Uro-Vaxom in recurrent urinary tract infection. The German Urinary Tract Infection Study Group. , 1990, British journal of urology.

[56]  J. G. Michael,et al.  Cationization of protein antigens. II. Alteration of regulatory properties. , 1987, Journal of immunology.

[57]  C. Bolin,et al.  Passive immunization with antibodies against iron-regulated outer membrane proteins protects turkeys from Escherichia coli septicemia , 1987, Infection and immunity.

[58]  J. G. Michael,et al.  Cationization of protein antigens. I. Alteration of immunogenic properties. , 1987, Journal of immunology.

[59]  J. G. Michael,et al.  Cationic antigens. Problems associated with measurement by ELISA. , 1986, Journal of immunological methods.

[60]  R. Freter,et al.  Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin , 1983, Infection and immunity.

[61]  E. Kamil,et al.  Induction of membranous nephropathy in rabbits by administration of an exogenous cationic antigen. , 1982, The Journal of clinical investigation.