Novel mouse mutants with primary cellular immunodeficiencies generated by genome-wide mutagenesis

J Allergy Clin Immunol. 2008 Jan;121(1):179-184.e7. doi: 10.1016/j.jaci.2007.07.018. Epub 2007 Sep 4.

Abstract

Background: Primary cellular immunodeficiencies are a group of genetic disorders in which 1 or more components of the cellular immune system are lacking or dysfunctional.

Objective: We sought to identify novel mouse mutants that display primary cellular immunodeficiencies.

Methods: Genome-wide N-ethyl-N-nitrosourea mutagenesis was performed in mice, followed by a phenotype screen of immunologic blood parameters.

Results: We identified novel mouse mutants with isolated B-cell deficiency, combined block in early B- and T-cell development, combined T-cell and natural killer cell reduction, and 3 different forms of T-cell deficiencies. One of the mutants, designated DeltaT3, displayed a combined phenotype of increased IgE, absence of peripheral T cells, and block in late thymocyte differentiation. In addition, DeltaT3 mice were unable to mount specific humoral immune responses. Chromosomal mapping and sequencing of candidate genes revealed a novel point mutation in the kinase domain of the T-cell receptor zeta chain-associated protein kinase (Zap70). In contrast to Zap70-deficient mice, DeltaT3 mutants displayed normal Zap70 mRNA and residual Zap70 protein levels. Complementation studies with Zap70-deficient mice confirmed that the point mutation found in Zap70 was causative for the DeltaT3 phenotype, including increased IgE plasma levels, a phenotype that has not been associated with altered Zap70 function in the past.

Conclusion: Random genome-wide mutagenesis combined with a phenotype screen can be used to generate novel mouse mutants with primary cellular immunodeficiencies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylating Agents*
  • Animals
  • Ethylnitrosourea*
  • Genome / genetics*
  • Immunoglobulin E / blood
  • Immunologic Deficiency Syndromes / genetics*
  • Immunologic Deficiency Syndromes / pathology
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Mice, Mutant Strains*
  • Mutagenesis*
  • Phenotype
  • ZAP-70 Protein-Tyrosine Kinase / genetics

Substances

  • Alkylating Agents
  • Immunoglobulin E
  • ZAP-70 Protein-Tyrosine Kinase
  • Zap70 protein, mouse
  • Ethylnitrosourea