Alternative splice forms of CTLA-4 induced by antisense mediated splice-switching influences autoimmune diabetes susceptibility in NOD mice

Nucleic Acid Ther. 2014 Apr;24(2):114-26. doi: 10.1089/nat.2013.0449. Epub 2014 Feb 4.


Activated and regulatory T cells express the negative co-stimulatory molecule cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) that binds B7 on antigen-presenting cells to mediate cellular responses. Single nucleotide polymorphisms in the CTLA-4 gene have been found to affect alternative splicing and are linked to autoimmune disease susceptibility or resistance. Increased expression of a soluble splice form (sCTLA-4), lacking the transmembrane domain encoded by exon 3, has been shown to accelerate autoimmune pathology. In contrast, an exon 2-deficient form lacking the B7 ligand binding domain (liCTLA-4), expressed by diabetes resistant mouse strains has been shown to be protective when expressed as a transgene in diabetes susceptible non-obese diabetic (NOD) mice. We sought to employ an antisense-targeted splice-switching approach to independently produce these CTLA-4 splice forms in NOD mouse T cells and observe their relative impact on spontaneous autoimmune diabetes susceptibility. In vitro antisense targeting of the splice acceptor site for exon 2 produced liCTLA-4 while targeting exon 3 produced the sCTLA-4 form in NOD T cells. The liCTLA-4 expressing T cells exhibited reduced activation, proliferation and increased adhesion to intercellular adhesion molecule-1 (ICAM-1) similar to treatment with agonist α-CTLA-4. Mice treated to produce liCTLA-4 at the time of elevated blood glucose levels exhibited a significant reduction in the incidence of insulitis and diabetes, whereas a marked increase in the incidence of both was observed in animals treated to produce sCTLA-4. These findings provide further support that alternative splice forms of CTLA-4 affects diabetes susceptibility in NOD mice and demonstrates the therapeutic utility of antisense mediated splice-switching for modulating immune responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abatacept
  • Alternative Splicing
  • Amino Acid Sequence
  • Animals
  • Autoimmunity / genetics*
  • Base Sequence
  • CTLA-4 Antigen / antagonists & inhibitors
  • CTLA-4 Antigen / genetics*
  • CTLA-4 Antigen / immunology
  • Cell Adhesion
  • Cell Proliferation
  • Diabetes Mellitus, Type 1 / genetics*
  • Diabetes Mellitus, Type 1 / immunology
  • Diabetes Mellitus, Type 1 / pathology
  • Disease Susceptibility / immunology*
  • Exons
  • Gene Expression Regulation
  • Immunoconjugates / pharmacology
  • Intercellular Adhesion Molecule-1 / genetics
  • Intercellular Adhesion Molecule-1 / immunology
  • Lymphocyte Activation
  • Mice
  • Mice, Inbred NOD
  • Molecular Sequence Data
  • Oligonucleotides, Antisense / genetics*
  • Oligonucleotides, Antisense / immunology
  • Protein Isoforms / antagonists & inhibitors
  • Protein Isoforms / genetics
  • Protein Isoforms / immunology
  • Severity of Illness Index
  • T-Lymphocytes, Regulatory / drug effects
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / pathology


  • CTLA-4 Antigen
  • Immunoconjugates
  • Oligonucleotides, Antisense
  • Protein Isoforms
  • Intercellular Adhesion Molecule-1
  • Abatacept