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Helicobacter pylori exploits human CEACAMs via HopQ for adherence and translocation of CagA

Nature Microbiology volume 2, Article number: 16188 (2017) Cite this article

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An Erratum to this article was published on 31 October 2016

Abstract

Helicobacter pylori (Hp) strains that carry the cag type IV secretion system (cag-T4SS) to inject the cytotoxin-associated antigen A (CagA) into host cells are associated with peptic ulcer disease and gastric adenocarcinoma. CagA translocation by Hp is mediated by β1 integrin interaction of the cag-T4SS. However, other cellular receptors or bacterial outer membrane adhesins essential for this process are unknown. Here, we identify the HopQ protein as a genuine Hp adhesin, exploiting defined members of the carcinoembryonic antigen-related cell adhesion molecule family (CEACAMs) as host cell receptors. HopQ binds the amino-terminal IgV-like domain of human CEACAM1, CEACAM3, CEACAM5 or CEACAM6 proteins, thereby enabling translocation of the major pathogenicity factor CagA into host cells. The HopQ–CEACAM interaction is characterized by a remarkably high affinity (KD from 23 to 268 nM), which is independent of CEACAM glycosylation, identifying CEACAMs as bona fide protein receptors for Hp. Our data suggest that the HopQ–CEACAM interaction contributes to gastric colonization or Hp-induced pathologies, although the precise role and functional consequences of this interaction in vivo remain to be determined.

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Figure 1: Hp interacts with soluble CEACAM1 and CEACAM5 molecules with different specificities.
Figure 2: The Hp outer membrane protein HopQ is the adhesin interacting with soluble CEACAM1 and CEACAM5.
Figure 3: The HopQ–CEACAM interaction is essential for Hp CagA translocation.
Figure 4: CEACAM1, CEACAM3, CEACAM5 and CEACAM6 act as cellular receptors for Hp, and the N-terminal IgV domain of CEACAM5 is sufficient to mediate CagA translocation.
Figure 5: Hp co-localizes with CEACAM5 in the gastric tissue of Hp-infected patients.
Figure 6: CEACAM5 expression levels determined on human gastric biopsy sections.

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Acknowledgements

The authors acknowledge support from collaborating gastroenterologists who collected gastric biopsies in the framework of the Africa Infectiology Study in Nigeria, especially at Lagos University Teaching Hospital, the University College Hospital (UCH) Ibadan and the University Teaching Hospital Complex Ile-Ife. The authors thank V. Naegele for CEACAM1 and CEACAM5 transfected HEK293 cells, E. Vetter for staining of CEA immunohistology sections, M. Schiemann and L. Henkel for FACS sorting of transfected cells and E. Weiss for technical support. This work was supported by grants from DFG (HA2856/6-2 to C.R.H. and HA2697/16-1, 17-1 and 18-1 and SFB914 Project B05 to R.H.) and in part by an Alexander von Humboldt Foundation Experienced Research Fellowship (to E.J.S.).

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Author notes

  1. Verena Königer and Lea Holsten: These authors contributed equally to this work.

Authors and Affiliations

  1. Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstraße 9a, D-80336 München, Germany

    Verena Königer, Lea Holsten, Ute Harrison, Benjamin Busch, Eva Loell, Qing Zhao, Wolfgang Fischer & Rainer Haas

  2. Institute of Human Virology, University of Maryland School of Medicine, University of Maryland, Baltimore, Maryland 21201, USA

    Daniel A. Bonsor & Eric J. Sundberg

  3. Lehrstuhl Zellbiologie, Fachbereich Biologie, Universität Konstanz, D-78457 Konstanz, Germany

    Alexandra Roth, Arnaud Kengmo-Tchoupa & Christof R. Hauck

  4. Molecular Biology and Biotechnology Division, Nigerian Institute of Medical Research, 6 Edmond Crescent, Yaba, PMB 2013 Lagos, Nigeria

    Stella I. Smith

  5. Institut für Pathologie, Ludwig-Maximilians-Universität, D-80337 München, Germany

    Susanna Mueller

  6. Department of Medicine, University of Maryland School of Medicine, University of Maryland, Baltimore, 21201, Maryland, USA

    Eric J. Sundberg

  7. Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland, Baltimore, 21201, Maryland, USA

    Eric J. Sundberg

  8. Tumor Immunology Laboratory, LIFE Center, Ludwig-Maximilians-Universität, Feodor-Lynen-Str. 19, D-81377 München, Germany

    Wolfgang Zimmermann

  9. Department of Urology, University Hospital, Ludwig-Maximilians-Universität, Marchioninistr. 15, D-81377 München, Germany

    Wolfgang Zimmermann

  10. German Center for Infection Research (DZIF), partner site LMU München, Germany

    Rainer Haas

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Contributions

V.K. carried out Hp mutant generation, pulldown assays and knockdowns. L.H. performed mutant CEACAM construction and Hp binding assays. E.L. carried out microscopy studies with patient material and B.B. carried out microscopy studies with Hp-infected cell lines. D.A.B. investigated the expression of HopQ and CEA-N in E. coli and carried out ITC experiments. U.H. performed in vivo studies and histological experiments. A.R. generated the soluble CEACAM construct. A.K.-T. prepared and purified soluble CEACAM constructs. S.I.S. coordinated biopsies in Lagos. S.M. generated pathology data. E.J.S. analysed the ITC experimental results. W.Z. carried out CEACAM plasmid construction. Q.Z. conducted CEACAM5 luciferase reporter assays. W.F. carried out Hp omp gene analysis and Hp mutant construction. C.R.H. provided CEACAM reagents and CEA-transgenic mice. R.H. designed and coordinated the project and wrote the paper.

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Correspondence to Rainer Haas.

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The authors declare no competing financial interests.

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Supplementary Figures 1-10, Supplementary Table 1-4, Original gel images (PDF 4790 kb)

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Königer, V., Holsten, L., Harrison, U. et al. Helicobacter pylori exploits human CEACAMs via HopQ for adherence and translocation of CagA. Nat Microbiol 2, 16188 (2017). https://doi.org/10.1038/nmicrobiol.2016.188

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