Hypomorphic Rag1 mutations alter the preimmune repertoire at early stages of lymphoid development

Blood. 2018 Jul 19;132(3):281-292. doi: 10.1182/blood-2017-12-820985. Epub 2018 May 9.

Abstract

Hypomorphic RAG1 mutations allowing residual T- and B-cell development have been found in patients presenting with delayed-onset combined immune deficiency with granulomas and/or autoimmunity (CID-G/AI) and abnormalities of the peripheral T- and B-cell repertoire. To examine how hypomorphic Rag1 mutations affect the earliest stages of lymphocyte development, we used CRISPR/Cas9 to generate mouse models with mutations equivalent to those found in patients with CID-G/AI. Immunological characterization showed partial development of T and B lymphocytes, with persistence of naïve cells and preserved serum immunoglobulin but impaired antibody responses and presence of autoantibodies, thereby recapitulating the phenotype seen in patients with CID-G/AI. By using high-throughput sequencing, we identified marked skewing of Igh V and Trb V gene usage in early progenitors, with a bias for productive Igh and Trb rearrangements after selection occurred and increased apoptosis of B-cell progenitors. Rearrangement at the Igk locus was impaired, and polyreactive immunoglobulin M antibodies were detected. This study provides novel insights into how hypomorphic Rag1 mutations alter the primary repertoire of T and B cells, setting the stage for immune dysregulation frequently seen in patients.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Alleles
  • Animals
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • Biomarkers
  • Cell Differentiation / genetics*
  • Disease Susceptibility / immunology
  • Gene Editing
  • Gene Expression Regulation
  • Genes, RAG-1*
  • Genetic Predisposition to Disease
  • Immunity, Humoral
  • Immunophenotyping
  • Lymphopoiesis / genetics*
  • Mice
  • Mice, Transgenic
  • Mutation*
  • Phenotype
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • V(D)J Recombination

Substances

  • Biomarkers