New insights into the cytokine spectrum in Graves' disease using an autoimmune mouse model with hyperthyroidism.

Introduction

Endocrine orbitopathy (EO) is the most common extra-thyroidal manifestation of Graves' disease, an autoimmune disease mediated by autoantibodies against the TSH receptor. We developed and established an experimental EO mouse model by genetically immunizing BALB/c mice with a plasmid encoding the A subunit of human TSHR. Cytokines seem to have a strong influence on the development of endocrine orbitopathy. The aim was to analyze these in more detail.

Material and Methods

The retroorbital tissue from female BALB/c mice immunized with the plasmid encoding the TSHR-A subunit or with the control ß-Gal plasmid, the following studies were performed: immunohistochemical analysis of F4/80, CD3, and TNF-α; macrophage migration; and cytokine analyses by Legend Plex and ELISA.

Results

The retroorbital tissue showed an inflammatory reaction with macrophages and CD3-positive T cells as well as TNF-α positive cells in the tissue. A migration assay showed, after stimulation with TNF-α, an increased number of macrophages in mouse orbital fibroblasts (mOF's). Furthermore, stimulation with TNF-α resulted in increased levels of CCL-2, CCL-5, and CCL-20, as well as CXCL-10 and CXCL-30 in TSHR-immunized mice.

Discussion

In the present study the cytokine spectrum in mOFs of EO mice and control mice was analyzed for the first time. CCL-2, CCL-5 and CCL-20 as well as CXCL-10 and CXCL-30 seem to have an influence on the development of endocrine orbitopathy.

Publication History

Article published online:
24 May 2022