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Horm Metab Res 2013; 45(06): 401-407
DOI: 10.1055/s-0032-1333224
Original Basic
© Georg Thieme Verlag KG Stuttgart · New York

Examination of Orbital Tissues in Murine Models of Graves’ Disease Reveals Expression of UCP-1 and the TSHR in Retrobulbar Adipose Tissues

K.T. M. Johnson
1   Department for Ophthalmology, University of Duisburg-Essen, Essen, Germany
,
B. Wiesweg
1   Department for Ophthalmology, University of Duisburg-Essen, Essen, Germany
,
M. Schott
2   Department of Endocrinology and Diabetology, University of Düsseldorf, Düsseldorf, Germany
,
M. Ehlers
2   Department of Endocrinology and Diabetology, University of Düsseldorf, Düsseldorf, Germany
,
M. Müller
1   Department for Ophthalmology, University of Duisburg-Essen, Essen, Germany
,
W. B. Minich
3   Institute for Experimental Endocrinology – EnForCé, Charité, Berlin, ­Germany
,
Y. Nagayama
4   Department of Molecular Medicine, Atomic Bomb Disease Institute, Japan University Graduate School of Biomedical Sciences, Nagasaki, Japan
,
E. Gulbins
5   Department of Molecular Biology, University of Duisburg-Essen, Essen, Germany
,
A. K. Eckstein
1   Department for Ophthalmology, University of Duisburg-Essen, Essen, Germany
,
U. Berchner-Pfannschmidt
1   Department for Ophthalmology, University of Duisburg-Essen, Essen, Germany
› Author Affiliations
Further Information

Publication History

received 04 June 2012

accepted 10 December 2012

Publication Date:
05 February 2013 (online)

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Abstract

Over the past decade a number of murine models of Graves’ disease (GD) have been described. The full symptom complex, including typical orbital changes, however, could not yet be induced. In this report, we examined the influence of modified immunization protocols on orbital pathology. C57BL/6 and BALB/c mice were immunized against the human TSH receptor (TSHR), using either a TSHR encoding plasmid or a TSHR A-subunit adenovirus. Prior to immunization with the TSHR plasmid, regulatory T cells were depleted in one group of each strain. TSHR-stimulating antibodies (TSAbs) were evaluated and orbits were stained immunohistochemically for F4/80, uncoupling protein-1 (UCP-1) and the TSHR. We found that after depletion of regulatory T cells, incidence of TSAb was increased in TSHR plasmid immunized C57BL/6 mice. Examination of early immunized mice showed no antibody production. However, a TSHR epitope-specific cellular immune response could be detected by tetramer-analyses. Adenoviral immunization lead to TSAb production in all but one animal. Analysis of F4/80 positive cells in retrobulbar fat revealed no significant macrophage infiltration in the orbits of immunized mice. Immunohistochemical staining shows co-localization of F4/80 positive cells, UCP-1 and the TSHR in retrobulbar fat. Though targets for TSHR autoimmunity could clearly be shown, immunization methods were not efficient enough to cause clear signs of orbital inflammation.

Key words

Graves’ orbitopathy - animal model - brown adipose tissue - macrophages
 

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