Comparative Assessment of Female Mouse Model of Graves' Orbitopathy Under Different Environments, Accompanied by Proinflammatory Cytokine and T-Cell Responses to Thyrotropin Hormone Receptor Antigen

Endocrinology. 2016 Apr;157(4):1673-82. doi: 10.1210/en.2015-1829. Epub 2016 Feb 12.

Abstract

We recently described a preclinical model of Graves' orbitopathy (GO), induced by genetic immunization of eukaryotic expression plasmid encoding human TSH receptor (TSHR) A-subunit by muscle electroporation in female BALB/c mice. The onset of orbital pathology is characterized by muscle inflammation, adipogenesis, and fibrosis. Animal models of autoimmunity are influenced by their environmental exposures. This follow-up study was undertaken to investigate the development of experimental GO in 2 different locations, run in parallel under comparable housing conditions. Functional antibodies to TSHR were induced in TSHR A-subunit plasmid-immunized animals, and antibodies to IGF-1 receptor α-subunit were also present, whereas control animals were negative in both locations. Splenic T cells from TSHR A-subunit primed animals undergoing GO in both locations showed proliferative responses to purified TSHR antigen and secreted interferon-γ, IL-10, IL-6, and TNF-α cytokines. Histopathological evaluation showed orbital tissue damage in mice undergoing GO, manifest by adipogenesis, fibrosis, and muscle damage with classic signs of myopathy. Although no inflammatory infiltrate was observed in orbital tissue in either location, the appearances were consistent with a "hit-and-run" immune-mediated inflammatory event. A statistically significant increase of cumulative incidence of orbital pathology when compared with control animals was shown for both locations, confirming onset of orbital dysimmune myopathy. Our findings confirm expansion of the model in different environments, accompanied with increased prevalence of T cell-derived proinflammatory cytokines, with relevance for pathogenesis. Wider availability of the model makes it suitable for mechanistic studies into pathogenesis and undertaking of novel therapeutic approaches.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens / immunology
  • CD3 Complex / immunology
  • CD3 Complex / metabolism
  • Cell Proliferation
  • Cytokines / immunology*
  • Cytokines / metabolism
  • Disease Models, Animal*
  • Female
  • Graves Ophthalmopathy / immunology*
  • Graves Ophthalmopathy / metabolism
  • Humans
  • Immunohistochemistry
  • Inflammation Mediators / immunology*
  • Inflammation Mediators / metabolism
  • Interferon-gamma / immunology
  • Interferon-gamma / metabolism
  • Interleukin-10 / immunology
  • Interleukin-10 / metabolism
  • Interleukin-6 / immunology
  • Interleukin-6 / metabolism
  • Mice, Inbred BALB C
  • Receptors, Thyrotropin / immunology*
  • Receptors, Thyrotropin / metabolism
  • Spleen / immunology
  • Spleen / metabolism
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • Tumor Necrosis Factor-alpha / immunology
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Antigens
  • CD3 Complex
  • Cytokines
  • Inflammation Mediators
  • Interleukin-6
  • Receptors, Thyrotropin
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Interferon-gamma