A high-throughput sequencing approach identifies immunotherapeutic targets for bacterial meningitis in neonatesResearch in context
Stéphanie Pons,
Eric Frapy,
Youssouf Sereme,
Charlotte Gaultier,
François Lebreton,
Andrea Kropec,
Olga Danilchanka,
Laura Schlemmer,
Cécile Schrimpf,
Margaux Allain,
François Angoulvant,
Hervé Lecuyer,
Stéphane Bonacorsi,
Hugues Aschard,
Harry Sokol,
Colette Cywes-Bentley,
John J. Mekalanos,
Thomas Guillard,
Gerald B. Pier,
Damien Roux,
David Skurnik
Affiliations
Stéphanie Pons
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Department of Anesthesiology and Critical Care, Sorbonne University, GRC 29, AP-HP, DMU DREAM, Pitié-Salpêtrière, Paris, France
Eric Frapy
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France; Faculté de Médecine, University of Paris City, Paris, France
Youssouf Sereme
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France; Faculté de Médecine, University of Paris City, Paris, France
Charlotte Gaultier
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
François Lebreton
Department of Ophthalmology and Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02114, USA
Andrea Kropec
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
Olga Danilchanka
Department of Microbiology, Harvard Medical School, Boston, MA 02115, USA
Laura Schlemmer
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France
Cécile Schrimpf
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France
Margaux Allain
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France
François Angoulvant
Assistance Publique - Hôpitaux de Paris, Pediatric Emergency Department, Necker-Enfants Malades University Hospital, University of Paris City, Paris, France; INSERM, Centre de Recherche des Cordeliers, UMRS 1138, Sorbonne Université, Université de Paris, Paris, France
Hervé Lecuyer
CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France; Faculté de Médecine, University of Paris City, Paris, France; Department of Clinical Microbiology, Fédération Hospitalo-Universitaire Prématurité (FHU PREMA), Necker-Enfants Malades University Hospital, University of Paris City, Paris, France
Stéphane Bonacorsi
E IAME, UMR 1137, INSERM, Université de Paris, AP-HP, Paris, France; Laboratoire de Microbiologie, Hôpital Robert Debré, AP-HP, Paris, France
Hugues Aschard
Centre de Bioinformatique, Biostatistique et Biologie Intégrative (C3BI), Institut Pasteur, Paris, France; Department of Epidemiology, Harvard TH Chan School of Public Health, Boston, MA, USA
Harry Sokol
Gastroenterology Department, Sorbonne University, INSERM, Centre de Recherche Saint-Antoine, CRSA, AP-HP, Saint Antoine Hospital, F-75012 Paris, France; INRA, UMR1319 Micalis & AgroParisTech, Jouy en Josas, France; Paris Centre for Microbiome Medicine FHU, Paris, France
Colette Cywes-Bentley
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
John J. Mekalanos
Department of Microbiology, Harvard Medical School, Boston, MA 02115, USA
Thomas Guillard
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Université de Reims Champagne-Ardenne, SFR CAP-Santé, Inserm UMR-S 1250 P3Cell, Reims, France; Laboratoire de Bactériologie-Virologie-Hygiène Hospitalière-Parasitologie-Mycologie, CHU, Reims, France
Gerald B. Pier
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
Damien Roux
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Université de Paris, INSERM, UMR 1137 IAME, F-75018 Paris, France; AP-HP, Médecine Intensive Réanimation, Hôpital Louis Mourier, F-92700 Colombes, France
David Skurnik
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; CNRS, INSERM, Institut Necker Enfants Malades-INEM, F-75015 Paris, France; Faculté de Médecine, University of Paris City, Paris, France; Department of Clinical Microbiology, Fédération Hospitalo-Universitaire Prématurité (FHU PREMA), Necker-Enfants Malades University Hospital, University of Paris City, Paris, France; Corresponding author. Département de Microbiologie, Université de Paris- Hôpital Necker Enfants Malades, 149, rue de Sèvres, 75015 Paris, France.
Summary: Background: Worldwide, Escherichia coli is the leading cause of neonatal Gram-negative bacterial meningitis, but full understanding of the pathogenesis of this disease is not yet achieved. Moreover, to date, no vaccine is available against bacterial neonatal meningitis. Methods: Here, we used Transposon Sequencing of saturated banks of mutants (TnSeq) to evaluate E. coli K1 genetic fitness in murine neonatal meningitis. We identified E. coli K1 genes encoding for factors important for systemic dissemination and brain infection, and focused on products with a likely outer-membrane or extra-cellular localization, as these are potential vaccine candidates. We used in vitro and in vivo models to study the efficacy of active and passive immunization. Results: We selected for further study the conserved surface polysaccharide Poly-β-(1-6)-N-Acetyl Glucosamine (PNAG), as a strong candidate for vaccine development. We found that PNAG was a virulence factor in our animal model. We showed that both passive and active immunization successfully prevented and/or treated meningitis caused by E. coli K1 in neonatal mice. We found an excellent opsonophagocytic killing activity of the antibodies to PNAG and in vitro these antibodies were also able to decrease binding, invasion and crossing of E. coli K1 through two blood brain barrier cell lines. Finally, to reinforce the potential of PNAG as a vaccine candidate in bacterial neonatal meningitis, we demonstrated that Group B Streptococcus, the main cause of neonatal meningitis in developed countries, also produced PNAG and that antibodies to PNAG could protect in vitro and in vivo against this major neonatal pathogen. Interpretation: Altogether, these results indicate the utility of a high-throughput DNA sequencing method to identify potential immunotherapy targets for a pathogen, including in this study a potential broad-spectrum target for prevention of neonatal bacterial infections. Fundings: ANR Seq-N-Vaq, Charles Hood Foundation, Hearst Foundation, and Groupe Pasteur Mutualité.
