A systematic analysis of recombination activity and genotype-phenotype correlation in human recombination-activating gene 1 deficiency - 29/03/14
, Luigi D. Notarangelo, MD a, ⁎ 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.
Le texte complet de cet article est disponible en PDF.Key words : Recombination-activating gene 1, V(D)J recombination, severe combined immune deficiency, Omenn syndrome, autoimmunity, genotype-phenotype correlation, immune repertoire
Abbreviations used : A-MuLV, CDR, CID-G/A, γδ-T, GFP, HBR, hRAG1, ICL, mRag1, NBR, OS, PON-P, RAG, RF, RSS, SCID, TCR
Plan
| Supported in part by National Institutes of Health grants U54AI082973 (to L.D.N.) and 1P01AI076210-01A1 (to L.D.N., F.W.A., and R.S.G.). F.W.A is an investigator of the Howard Hughes Medical Institute. This work received additionally contributions from the March of Dimes (grant 1-FY-13-500 to L.D.N.), the Jeffrey Modell Foundation (to L.D.N.), the Translational Research Program grant TRP 2009 042809 (to L.D.N.), the Dubai Harvard Foundation for Medical Research (to L.D.N. and R.S.G.), and the Manton Foundation (to Y.N.L. and L.D.N.). |
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| Disclosure of potential conflict of interest: Y. N. Lee has received research support from the Manton Foundation and the March of Dimes and is employed by Boston Children's Hospital. F. Frugoni, J. Puck, and F. Alt have received research support from the National Institutes of Health (NIH). J. E. Walter is employed by Massachusetts General Hospital and receives research support from the National Institute of Allergy and Infectious Disease. L. A. Henderson has received travel support from the American College of Rheumatology and the American Academy of Pediatrics. S.-Y. Pai is employed by Boston Children's Hospital and receives research support from the Translational Investigator Service Award from Boston Children's Hospital. P. Soler-Palacin has consultant arrangements with CSL Behring; has provided expert testimony on behalf of CSL Behring; has received research support from Baxter, CSL Behring, and Octapharma; has received payment for lectures from CSL Behring; and has received travel support from the European Society of Immunodeficiencies. R. S. Geha has received research support from the NIH and the Dubai Harvard Foundation for Medical Research. L. D. Notarangelo has received research support from the NIH, the Jeffrey Modell Foundation, the March of Dimes, the Dubai Harvard Foundation for Medical Research, and the Manton Foundation; has board memberships with the Program in Molecular and Cellular Medicine and Pediatric University Hospital “Meyer” in Florence, Italy; is employed by Boston Children's Hospital; and has received royalties from UpToDate. The rest of the authors declare that they have no relevant conflicts of interest. |
Vol 133 - N° 4
P. 1099 - avril 2014 Retour au numéro
Article précédent
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