Aurélie Berland1, Jérémie Rosain2, Sophie Kaltenbach1, Vincent Allain3, Nizar Mahlaoui4, Isabelle Melki5, Alice Fievet6, Catherine Dubois d'Enghien7, Marie Ouachée-Chardin8, Laurence Perrin9, Nathalie Auger10, Funda Erol Cipe11, Andrea Finocchi12, Figen Dogu13, Felipe Suarez14, Despina Moshous15, Thierry Leblanc8, Alexandre Belot16, Claire Fieschi17, David Boutboul17, Marion Malphettes17, Lionel Galicier17, Eric Oksenhendler17, Stéphane Blanche18, Alain Fischer19, Patrick Revy1, Dominique Stoppa-Lyonnet20, Capucine Picard2, Jean-Pierre de Villartay21. 1. Laboratory "Genome Dynamics in the Immune System", INSERM UMR1163, Paris, France; Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France. 2. Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France; Study Center for Primary Immunodeficiencies, Necker-Enfants Malades Hospital, Assistance Publique Hôpitaux de Paris (APHP), Necker Medical School, Paris, France. 3. Study Center for Primary Immunodeficiencies, Necker-Enfants Malades Hospital, Assistance Publique Hôpitaux de Paris (APHP), Necker Medical School, Paris, France. 4. Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France; Pediatric Immuno-Haematology and Rheumatology Unit, Necker Enfants Malades University Hospital, Assistance Publique-Hôpitaux de Paris, Paris, France. 5. Pediatric Immuno-Haematology and Rheumatology Unit, Necker Enfants Malades University Hospital, Assistance Publique-Hôpitaux de Paris, Paris, France; General Pediatrics, Infectious Disease and Internal Medicine Department, Hôpital Robert Debré, (APHP), Paris, France. 6. INSERM U830, Institut Curie, Paris, France; Service de Génétique, Institut Curie, Paris, France. 7. Service de Génétique, Institut Curie, Paris, France. 8. Department of Pediatric Hematology, Robert-Debré (APHP), Paris, France. 9. Department of Genetics, Robert Debré Hospital, (APHP), Paris, France. 10. Department of Biopathology, Institut Gustave Roussy, Villejuif, France. 11. Department of Pediatric Allergy-Immunology, Kanuni Sultan Suleyman Research and Training Hospital, Istanbul, Turkey. 12. DPUO, University Department of Pediatrics, Bambino Gesù Children's Hospital and University of Tor Vergata School of Medicine, Rome, Italy. 13. Department of Pediatric Immunology and Allergy, Ankara University School of Medicine, Ankara, Turkey. 14. Department of Haematology, Necker-Enfants Malades University Hospital, (APHP), Paris, France. 15. Laboratory "Genome Dynamics in the Immune System", INSERM UMR1163, Paris, France; Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France; Pediatric Immuno-Haematology and Rheumatology Unit, Necker Enfants Malades University Hospital, Assistance Publique-Hôpitaux de Paris, Paris, France. 16. Pediatric Rheumatology, Nephrology and Dermatology Department, Hôpital Femme-Mère-Enfant, Hospices civils de Lyon, Lyon, France. 17. Department of Clinical Immunology, Hôpital Saint-Louis, (APHP), Paris, France. 18. Pediatric Immuno-Haematology and Rheumatology Unit, Necker Enfants Malades University Hospital, Assistance Publique-Hôpitaux de Paris, Paris, France. 19. Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France; Study Center for Primary Immunodeficiencies, Necker-Enfants Malades Hospital, Assistance Publique Hôpitaux de Paris (APHP), Necker Medical School, Paris, France; INSERM UMR1163, Paris, France; Collège de France, Paris, France. 20. INSERM U830, Institut Curie, Paris, France; Service de Génétique, Institut Curie, Paris, France; Université Paris Descartes Sorbonne Paris Cité, Paris, France. 21. Laboratory "Genome Dynamics in the Immune System", INSERM UMR1163, Paris, France; Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris, France. Electronic address: devillartay@gmail.com.
Abstract
BACKGROUND: V(D)J recombination ensures the diversity of the adaptive immune system. Although its complete defect causes severe combined immunodeficiency (ie, T-B- severe combined immunodeficiency), its suboptimal activity is associated with a broad spectrum of immune manifestations, such as late-onset combined immunodeficiency and autoimmunity. The earliest molecular diagnosis of these patients is required to adopt the best therapy strategy, particularly when it involves a myeloablative conditioning regimen for hematopoietic stem cell transplantation. OBJECTIVE: We aimed at developing biomarkers based on analysis of the T-cell receptor (TCR) α repertoire to assist in the diagnosis of patients with primary immunodeficiencies with V(D)J recombination and DNA repair deficiencies. METHODS: We used flow cytometric (fluorescence-activated cell sorting) analysis to quantify TCR-Vα7.2-expressing T lymphocytes in peripheral blood and developed PROMIDISα, a multiplex RT-PCR/next-generation sequencing assay, to evaluate a subset of the TCRα repertoire in T lymphocytes. RESULTS: The combined fluorescence-activated cell sorting and PROMIDISα analyses revealed specific signatures in patients with V(D)J recombination-defective primary immunodeficiencies or ataxia telangiectasia/Nijmegen breakage syndromes. CONCLUSION: Analysis of the TCRα repertoire is particularly appropriate in a prospective way to identify patients with partial immune defects caused by suboptimal V(D)J recombination activity, a DNA repair defect, or both. It also constitutes a valuable tool for the retrospective in vivo functional validation of variants identified through exome or panel sequencing. Its broader implementation might be of interest to assist early diagnosis of patients presenting with hypomorphic DNA repair defects inclined to experience acute toxicity during prehematopoietic stem cell transplantation conditioning.
BACKGROUND: V(D)J recombination ensures the diversity of the adaptive immune system. Although its complete defect causes severe combined immunodeficiency (ie, T-B- severe combined immunodeficiency), its suboptimal activity is associated with a broad spectrum of immune manifestations, such as late-onset combined immunodeficiency and autoimmunity. The earliest molecular diagnosis of these patients is required to adopt the best therapy strategy, particularly when it involves a myeloablative conditioning regimen for hematopoietic stem cell transplantation. OBJECTIVE: We aimed at developing biomarkers based on analysis of the T-cell receptor (TCR) α repertoire to assist in the diagnosis of patients with primary immunodeficiencies with V(D)J recombination and DNA repair deficiencies. METHODS: We used flow cytometric (fluorescence-activated cell sorting) analysis to quantify TCR-Vα7.2-expressing T lymphocytes in peripheral blood and developed PROMIDISα, a multiplex RT-PCR/next-generation sequencing assay, to evaluate a subset of the TCRα repertoire in T lymphocytes. RESULTS: The combined fluorescence-activated cell sorting and PROMIDISα analyses revealed specific signatures in patients with V(D)J recombination-defective primary immunodeficiencies or ataxia telangiectasia/Nijmegen breakage syndromes. CONCLUSION: Analysis of the TCRα repertoire is particularly appropriate in a prospective way to identify patients with partial immune defects caused by suboptimal V(D)J recombination activity, a DNA repair defect, or both. It also constitutes a valuable tool for the retrospective in vivo functional validation of variants identified through exome or panel sequencing. Its broader implementation might be of interest to assist early diagnosis of patients presenting with hypomorphic DNA repair defects inclined to experience acute toxicity during prehematopoietic stem cell transplantation conditioning.
Authors: Emmanuel Martin; Norbert Minet; Anne-Claire Boschat; Sylvia Sanquer; Steicy Sobrino; Christelle Lenoir; Jean Pierre de Villartay; Maria Leite-de-Moraes; Capucine Picard; Claire Soudais; Tim Bourne; Sophie Hambleton; Stephen M Hughes; Robert F Wynn; Tracy A Briggs; Smita Patel; Monica G Lawrence; Alain Fischer; Peter D Arkwright; Sylvain Latour Journal: JCI Insight Date: 2020-03-12
Authors: Jocelyn R Farmer; Zsofia Foldvari; Boglarka Ujhazi; Suk See De Ravin; Karin Chen; Jack J H Bleesing; Catharina Schuetz; Waleed Al-Herz; Roshini S Abraham; Avni Y Joshi; Beatriz T Costa-Carvalho; David Buchbinder; Claire Booth; Andreas Reiff; Polly J Ferguson; Asghar Aghamohammadi; Hassan Abolhassani; Jennifer M Puck; Mehdi Adeli; Caterina Cancrini; Paolo Palma; Alice Bertaina; Franco Locatelli; Gigliola Di Matteo; Raif S Geha; Maria G Kanariou; Lilia Lycopoulou; Marianna Tzanoudaki; John W Sleasman; Suhag Parikh; Gloria Pinero; Bernard M Fischer; Ghassan Dbaibo; Ekrem Unal; Turkan Patiroglu; Musa Karakukcu; Khulood Khalifa Al-Saad; Meredith A Dilley; Sung-Yun Pai; Cullen M Dutmer; Erwin W Gelfand; Christoph B Geier; Martha M Eibl; Hermann M Wolf; Lauren A Henderson; Melissa M Hazen; Carmem Bonfim; Beata Wolska-Kuśnierz; Manish J Butte; Joseph D Hernandez; Sarah K Nicholas; Polina Stepensky; Shanmuganathan Chandrakasan; Maurizio Miano; Emma Westermann-Clark; Vera Goda; Gergely Kriván; Steven M Holland; Olajumoke Fadugba; Sarah E Henrickson; Ahmet Ozen; Elif Karakoc-Aydiner; Safa Baris; Ayca Kiykim; Robbert Bredius; Birgit Hoeger; Kaan Boztug; Olga Pashchenko; Benedicte Neven; Despina Moshous; Jean-Pierre de Villartay; Ahmed Aziz Bousfiha; Harry R Hill; Luigi D Notarangelo; Jolan E Walter Journal: J Allergy Clin Immunol Pract Date: 2019-03-12
Authors: Ottavia M Delmonte; Jenna R E Bergerson; Tomoki Kawai; Hye Sun Kuehn; David H McDermott; Irene Cortese; Michael T Zimmermann; A Kerry Dobbs; Marita Bosticardo; Danielle Fink; Shamik Majumdar; Boaz Palterer; Francesca Pala; Nikita R Dsouza; Marie Pouzolles; Naomi Taylor; Katherine R Calvo; Stephen R Daley; Daniel Velez; Anahita Agharahimi; Katherine Myint-Hpu; Lesia K Dropulic; Jonathan J Lyons; Steven M Holland; Alexandra F Freeman; Rajarshi Ghosh; Morgan B Similuk; Julie E Niemela; Jennifer Stoddard; Douglas B Kuhns; Raul Urrutia; Sergio D Rosenzweig; Magdalena A Walkiewicz; Philip M Murphy; Luigi D Notarangelo Journal: Blood Date: 2021-09-23 Impact factor: 25.476
Authors: Cameron L Gardner; Mara Pavel-Dinu; Kerry Dobbs; Marita Bosticardo; Paul K Reardon; Justin Lack; Suk See DeRavin; Kent Le; Ezekiel Bello; Francesca Pala; Ottavia M Delmonte; Harry Malech; Amelie Montel-Hagan; Gay Crooks; Oreste Acuto; Matthew H Porteus; Luigi D Notarangelo Journal: J Clin Immunol Date: 2021-03-01 Impact factor: 8.542
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