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Germline CYBB mutations that selectively affect macrophages in kindreds with X-linked predisposition to tuberculous mycobacterial disease

Nature Immunology volume 12, pages 213–221 (2011)Cite this article

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Abstract

Germline mutations in CYBB, the human gene encoding the gp91phox subunit of the phagocyte NADPH oxidase, impair the respiratory burst of all types of phagocytes and result in X-linked chronic granulomatous disease (CGD). We report here two kindreds in which otherwise healthy male adults developed X-linked recessive Mendelian susceptibility to mycobacterial disease (MSMD) syndromes. These patients had previously unknown mutations in CYBB that resulted in an impaired respiratory burst in monocyte-derived macrophages but not in monocytes or granulocytes. The macrophage-specific functional consequences of the germline mutation resulted from cell-specific impairment in the assembly of the NADPH oxidase. This 'experiment of nature' indicates that CYBB is associated with MSMD and demonstrates that the respiratory burst in human macrophages is a crucial mechanism for protective immunity to tuberculous mycobacteria.

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Figure 1: CYBB mutations encoding Q231P and T178 substitutions in XR-MSMD-2.
Figure 2: NADPH oxidase activity in PMNs and monocytes.
Figure 3: NADPH oxidase activity in MDMs.
Figure 4: NADPH oxidase activity in EBV-B cells.
Figure 5: Expression of gp91phox and flavocytochrome b558.
Figure 6: Expression and function of mutant gp91phox in cell lines.

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Acknowledgements

We thank the family members for participating in this study; J. Curnutte (3-V Biosciences) for EBV-B cells from CGD controls; D. Roos (University of Amsterdam) for mAb 449; F. Morel (Grenoble University) for mAb 7A2; M. Quinn (Montana State University) for mAb 54.1; all members of the two branches of the laboratory of Human Genetics of Infectious Diseases for discussions; and T. Leclerc, Y. Rose, M. de Suremain, M. Kezadi, and N. Stull for technical assistance. Supported by Institut National de la Santé et de la Recherche Médicale (J.B.), the European Union (NEOTIM EEA05095KKA to J.B., and HOMITB HEALTH-F3-2008-200732), the March of Dimes (RO5050KK to J.B.), Fundação de Amparo a Pesquisa do Estado de São Paulo (A.C.-N.), Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (A.C.-N.), Fondation BNP-Paribas, Fondation Schlumberger, Institut Universitaire de France, Agence Nationale de Recherche, The Rockefeller University Center for Clinical and Translational Science (5UL1RR024143-03), The Rockefeller University, the National Institutes of Health (AI 079788 and DK54369 to P.E.N., and HL045635 to M.C.D.), the Riley Children's Foundation (M.C.D.) and the Howard Hughes Medical Institute (J.-L.C.).

Author information

Author notes

  1. Andres A Arias, Guillaume Vogt and Capucine Picard: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Human Genetics of Infectious Diseases, Necker Branch, Institut National de la Santé et de la Recherche Médicale, U980, Paris, France.,

    Jacinta Bustamante, Capucine Picard, Lizbeth Blancas Galicia, Audrey V Grant, Marjorie Hubeau, Ariane Chapgier, Ludovic de Beaucoudrey, Anne Puel, Jacqueline Feinberg, Lucile Jannière, Claire Fieschi, Stéphanie Boisson-Dupuis, Laurent Abel & Jean-Laurent Casanova

  2. Paris Descartes University, Necker Medical School, Paris, France

    Jacinta Bustamante, Capucine Picard, Lizbeth Blancas Galicia, Audrey V Grant, Marjorie Hubeau, Ariane Chapgier, Ludovic de Beaucoudrey, Anne Puel, Jacqueline Feinberg, Lucile Jannière, Claire Fieschi, Stéphanie Boisson-Dupuis, Laurent Abel & Jean-Laurent Casanova

  3. Department of Pediatrics, Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, Indiana, USA

    Andres A Arias, Christophe C Marchal, Ethan Valinetz & Mary C Dinauer

  4. St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, New York, USA

    Guillaume Vogt, Carolina Prando, Stéphanie Boisson-Dupuis, Laurent Abel & Jean-Laurent Casanova

  5. Center for the Study of Primary Immunodeficiencies, Assistance Publique–Hôpitaux de Paris AP-HP, Necker Hospital, Paris, France

    Capucine Picard

  6. Pediatric Hematology-Immunology Unit, Necker Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France

    Capucine Picard, Stéphane Blanche & Jean-Laurent Casanova

  7. National Institute of Pediatrics, Unit of Immunodeficiency, Mexico City, Mexico

    Lizbeth Blancas Galicia

  8. National Genotyping Center, Evry, France

    Céline Besse & Anne Boland

  9. Department of Internal Medicine, Nantes Hospital, Nantes, France

    Jean-Marie Brisseau

  10. Department of Infectious Diseases, AP-HP, Necker Hospital, Paris, France

    Olivier Lortholary

  11. Adult Immunopathology Unit, Saint Louis Hospital, Paris, France

    Claire Fieschi

  12. Department of Pathology, Institut National de la Santé et de la Recherche Médicale, U602, Ambroise Paré Hospital and Versailles Saint-Quentin-en-Yvelines University, Boulogne, France

    Jean-François Emile

  13. Department of Pediatrics, Prince Naif Center for Immunology Research, College of Medicine, King Saud University, Riyadh, Saudi Arabia

    Saleh Al-Muhsen & Jean-Laurent Casanova

  14. Department of Pathology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Bruce Woda

  15. Departments of Pediatrics and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Peter E Newburger

  16. Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil

    Antonio Condino-Neto

Authors
  1. Jacinta Bustamante
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  2. Andres A Arias
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  4. Capucine Picard
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  5. Lizbeth Blancas Galicia
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  6. Carolina Prando
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Contributions

J.B., L.A. and J.-L.C. designed the study and contributed intellectually to the experimental process; J.B. did most of the experiments under the supervision of J.-L.C.; A.A.A., C.C.M., E.V. and M.C.D. did the experiments with retroviral transduction of gp91phox into EBV-B, CHO and PLB-985 cells; G.V. made the nonretroviral CYBB vectors, infected macrophages with BCG and made intellectual contributions to various experiments; C. Picard contributed to the recruitment of patients and initiated the clinical investigation; L.B.G., C. Prando, L.J. and M.H. analyzed many controls; A.C. did quantitative RT-PCR; L.d.B. did bioinformatics analysis; J.-F.E. did histological analysis of lymph nodes; B.W. did immunoperoxidase staining; A.V.G., C.B. and A.B. provided data for linkage analysis; A.P., J.F. and S.B.-D. provided experimental advice about cell culture; J.-M.B., S.B., O.L. and C.F. contributed to the recruitment and follow-up of the patients and CGD controls; S.A.-M., P.N., A.C.-N. and M.C.D. provided CGD controls and intellectual guidance for the development of various assays; J.B. and J.-L.C. wrote the paper; and all authors commented on and discussed the paper. B.W., P.E.N., A.C.-N. and M.C.D. contributed equally to this work.

Corresponding author

Correspondence to Jean-Laurent Casanova.

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Bustamante, J., Arias, A., Vogt, G. et al. Germline CYBB mutations that selectively affect macrophages in kindreds with X-linked predisposition to tuberculous mycobacterial disease. Nat Immunol 12, 213–221 (2011). https://doi.org/10.1038/ni.1992

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