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J Allergy Clin Immunol. 2014 Apr;133(4):1099-108. doi: 10.1016/j.jaci.2013.10.007. Epub 2013 Nov 28.

A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency.

Author information

1
Division of Immunology and Manton Center for Orphan Disease Research, Children's Hospital, Harvard Medical School, Boston, Mass.
2
Division of Immunology and Manton Center for Orphan Disease Research, Children's Hospital, Harvard Medical School, Boston, Mass; Division of Pediatric Allergy/Immunology, Massachusetts General Hospital for Children, Boston, Mass.
3
A. Nocivelli Institute for Molecular Medicine, Pediatric Clinic, University of Brescia, and the Section of Genetics, Department of Pathology Spedali Civili, Brescia, Italy.
4
Department of Paediatric Immunology, Newcastle Upon Tyne Hospital, NHS Foundation Trust, United Kingdom and Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
5
Department of Pediatrics, Faculty of Medicine, Kuwait University, Kuwait City, Kuwait.
6
Department of Pediatrics and Department of Microbiology, Infectiology and Immunology, University of Montreal, CHU Sainte-Justine Research Center, Montreal, Quebec, Canada.
7
Division of Hematology/Oncology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
8
Division of Hematology-Oncology, Boston Children's Hospital, Boston, Mass.
9
Divisions of Hematology and Oncology, Indiana University School of Medicine, Indianapolis, Ind.
10
Department of Pediatric Allergy and Immunology, Children's Hospital, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
11
Paediatric Infectious Diseases and Immunodeficiencies Unit, Vall d'Hebron University Hospital, Barcelona, Spain.
12
Division of Allergy and Immunology, Ann & Robert H. Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, Ill.
13
Immunology Clinic, Levine Children's Hospital, Carolinas Medical Center, Charlotte, NC.
14
Department of Pediatrics, University of California San Francisco and UCSF Benioff Children's Hospital, San Francisco, Calif.
15
DPUO, University Department of Pediatrics, Bambino Gesù Children's Hospital and University of Tor Vergata School of Medicine, Rome, Italy.
16
Division of Allergy, Immunology & Rheumatology, Department of Pediatrics, University of Utah, Salt Lake City, Utah.
17
Department of Experimental Medical Science, Lund University, Lund, Sweden.
18
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Boston Children's Hospital, and the Department of Genetics, Harvard Medical School, Boston, Mass. Electronic address: alt@enders.tch.harvard.edu.
19
Division of Immunology and Manton Center for Orphan Disease Research, Children's Hospital, Harvard Medical School, Boston, Mass. Electronic address: luigi.notarangelo@childrens.harvard.edu.

Abstract

BACKGROUND:

The recombination-activating gene (RAG) 1/2 proteins play a critical role in the development of T and B cells by initiating the VDJ recombination process that leads to generation of a broad T-cell receptor (TCR) and B-cell receptor repertoire. Pathogenic mutations in the RAG1/2 genes result in various forms of primary immunodeficiency, ranging from T(-)B(-) severe combined immune deficiency to delayed-onset disease with granuloma formation, autoimmunity, or both. It is not clear what contributes to such heterogeneity of phenotypes.

OBJECTIVE:

We sought to investigate the molecular basis for phenotypic diversity presented in patients with various RAG1 mutations.

METHODS:

We have developed a flow cytometry-based assay that allows analysis of RAG recombination activity based on green fluorescent protein expression and have assessed the induction of the Ighc locus rearrangements in mouse Rag1(-/-) pro-B cells reconstituted with wild-type or mutant human RAG1 (hRAG1) using deep sequencing technology.

RESULTS:

Here we demonstrate correlation between defective recombination activity of hRAG1 mutant proteins and severity of the clinical and immunologic phenotype and provide insights on the molecular mechanisms accounting for such phenotypic diversity.

CONCLUSIONS:

Using a sensitive assay to measure the RAG1 activity level of 79 mutations in a physiologic setting, we demonstrate correlation between recombination activity of RAG1 mutants and the severity of clinical presentation and show that RAG1 mutants can induce specific abnormalities of the VDJ recombination process.

KEYWORDS:

Omenn syndrome; Recombination-activating gene 1; V(D)J recombination; autoimmunity; genotype-phenotype correlation; immune repertoire; severe combined immune deficiency

PMID:
24290284
PMCID:
PMC4005599
DOI:
10.1016/j.jaci.2013.10.007
[Indexed for MEDLINE]
Free PMC Article

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