Recombination activity of human recombination-activating gene 2 (RAG2) mutations and correlation with clinical phenotype.
Adolescent
B-Lymphocytes
/ physiology
Cell Line, Transformed
Child
Child, Preschool
DNA-Binding Proteins
/ genetics
Disease Progression
Female
Gene Knockdown Techniques
Genetic Association Studies
High-Throughput Nucleotide Sequencing
Humans
Immunoglobulin Heavy Chains
/ genetics
Infant
Infant, Newborn
Male
Mutation
/ genetics
Nuclear Proteins
/ genetics
Polymorphism, Genetic
Receptors, Antigen, B-Cell
/ genetics
Severe Combined Immunodeficiency
/ genetics
Omenn syndrome
Recombination-activating gene 2
VDJ recombination
autoimmunity
genotype-phenotype correlation
severe combined immunodeficiency
Journal
The Journal of allergy and clinical immunology
ISSN: 1097-6825
Titre abrégé: J Allergy Clin Immunol
Pays: United States
ID NLM: 1275002
Informations de publication
Date de publication:
02 2019
02 2019
Historique:
received:
24
07
2017
revised:
22
04
2018
accepted:
27
04
2018
pubmed:
18
5
2018
medline:
26
2
2020
entrez:
18
5
2018
Statut:
ppublish
Résumé
Mutations in recombination-activating gene (RAG) 1 and RAG2 are associated with a broad range of clinical and immunologic phenotypes in human subjects. Using a flow cytometry-based assay, we aimed to measure the recombinase activity of naturally occurring RAG2 mutant proteins and to correlate our results with the severity of the clinical and immunologic phenotype. Abelson virus-transformed Rag2 The RAG2 variants showed a wide range of recombination activity. Mutations associated with severe combined immunodeficiency and Omenn syndrome had significantly lower activity than those detected in patients with less severe clinical presentations. Four variants (P253R, F386L, N474S, and M502V) previously thought to be pathogenic were found to have wild-type levels of activity. Use of bicistronic vectors permitted us to assess more carefully the effect of compound heterozygous mutations, with good correlation between GFP expression and the number and diversity of Igh rearrangements. Our data support genotype-phenotype correlation in the setting of RAG2 deficiency. The assay described can be used to define the possible disease-causing role of novel RAG2 variants and might help predict the severity of the clinical phenotype.
Sections du résumé
BACKGROUND
Mutations in recombination-activating gene (RAG) 1 and RAG2 are associated with a broad range of clinical and immunologic phenotypes in human subjects.
OBJECTIVE
Using a flow cytometry-based assay, we aimed to measure the recombinase activity of naturally occurring RAG2 mutant proteins and to correlate our results with the severity of the clinical and immunologic phenotype.
METHODS
Abelson virus-transformed Rag2
RESULTS
The RAG2 variants showed a wide range of recombination activity. Mutations associated with severe combined immunodeficiency and Omenn syndrome had significantly lower activity than those detected in patients with less severe clinical presentations. Four variants (P253R, F386L, N474S, and M502V) previously thought to be pathogenic were found to have wild-type levels of activity. Use of bicistronic vectors permitted us to assess more carefully the effect of compound heterozygous mutations, with good correlation between GFP expression and the number and diversity of Igh rearrangements.
CONCLUSIONS
Our data support genotype-phenotype correlation in the setting of RAG2 deficiency. The assay described can be used to define the possible disease-causing role of novel RAG2 variants and might help predict the severity of the clinical phenotype.
Identifiants
pubmed: 29772310
pii: S0091-6749(18)30699-7
doi: 10.1016/j.jaci.2018.04.027
pmc: PMC6295349
mid: NIHMS982557
pii:
doi:
Substances chimiques
DNA-Binding Proteins
0
Immunoglobulin Heavy Chains
0
Nuclear Proteins
0
RAG2 protein, human
0
Receptors, Antigen, B-Cell
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
726-735Subventions
Organisme : NIAID NIH HHS
ID : R01 AI100887
Pays : United States
Organisme : Intramural NIH HHS
ID : Z99 AI999999
Pays : United States
Informations de copyright
Copyright © 2018 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.
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