Immunoglobulins from Graves' disease patients interact with different sites on TSH receptor/LH-CG receptor chimeras than either TSH or immunoglobulins from idiopathic myxedema patients

Tahara, Kazuo; Ban, Toshiaki; Minegishi, Takashi; Kohn, Leonard D.

doi:10.1016/0006-291x(91)91335-a

Cited by 135 publications

(61 citation statements)

References 14 publications

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“…4B). These data are essentially identical to those previously reported with human Graves' sera (24,25), suggesting the crucial role of the N-terminal region of TSHR ectodomain in both human and murine TSI.…”

Section: Resultssupporting

confidence: 90%

“…Our study with chimeric receptors suggests that the epitopes on TSHR recognized by TSI in our model appear very similar to those observed in human disease (24,25), suggesting the involvement of the N terminus of TSHR ectodomain in both human and murine TSI. This is in agreement with the studies by Kikuoka et al (37), who have shown the importance of the N terminus of TSHR ectodomain (domains A and B in our chimeras) in induction of Graves' hyperthyroidism with Shimojo's model.…”

Section: Discussionsupporting

confidence: 76%

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A Novel Murine Model of Graves’ Hyperthyroidism with Intramuscular Injection of Adenovirus Expressing the Thyrotropin Receptor

Nagayama¹,

Kita-Furuyama

Ando

et al. 2002

The Journal of Immunology

177

207

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In this work we report a novel method to efficiently induce a murine model of Graves’ hyperthyroidism. Inbred mice of different strains were immunized by i.m. injection with adenovirus expressing thyrotropin receptor (TSHR) or β-galactosidase (1 × 1011 particles/mouse, three times at 3-wk intervals) and followed up to 8 wk after the third immunization. Fifty-five percent of female and 33% of male BALB/c (H-2d) and 25% of female C57BL/6 (H-2b) mice developed Graves’-like hyperthyroidism with elevated serum thyroxine (T4) levels and positive anti-TSHR autoantibodies with thyroid-stimulating Ig (TSI) and TSH-binding inhibiting Ig (TBII) activities. In contrast, none of female CBA/J (H-2k), DBA/1J (H-2q), or SJL/J (H-2s) mice developed Graves’ hyperthyroidism or anti-TSHR autoantibodies except SJL/J, which showed strong TBII activities. There was a significant positive correlation between TSI values and T4 levels, but the correlations between T4 and TBII and between TSI and TBII were very weak. TSI activities in sera from hyperthyroid mice measured with some chimeric TSH/lutropin receptors suggested that their epitope(s) on TSHR appeared similar to those in patients with Graves’ disease. The thyroid glands from hyperthyroid mice displayed diffuse enlargement with hypertrophy and hypercellularity of follicular epithelia with occasional protrusion into the follicular lumen, characteristics of Graves’ hyperthyroidism. Decreased amounts of colloid were also observed. However, there was no inflammatory cell infiltration. Furthermore, extraocular muscles from hyperthyroid mice were normal. Thus, the highly efficient means that we now report to induce Graves’ hyperthyroidism in mice will be very useful for studying the pathogenesis of autoimmunity in Graves’ disease.

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Section: Resultssupporting

confidence: 90%

Section: Discussionsupporting

confidence: 76%

A Novel Murine Model of Graves’ Hyperthyroidism with Intramuscular Injection of Adenovirus Expressing the Thyrotropin Receptor

Nagayama¹,

Kita-Furuyama

Ando

et al. 2002

The Journal of Immunology

177

207

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“…CHO cells expressing hTSHR (26,27) were grown to confluence in 96-well plates in F-12 medium supplemented with 10% FBS. Before the assay, spent medium was removed, and cells were washed with fresh medium.…”

Section: Camp Assaymentioning

confidence: 99%

Absence of IL-4, and Not Suppression of the Th2 Response, Prevents Development of Experimental Autoimmune Graves’ Disease

Dogan

Vasu

Holterman

et al. 2003

The Journal of Immunology

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In autoimmune Graves’ disease (GD), autoantibodies bind to the thyrotropin receptor (TSHR) and cause hyperthyroidism. We studied the effects of fms-like tyrosine kinase receptor 3 ligand (Flt3-L) or GM-CSF treatment on the development of experimental autoimmune GD (EAGD) in mice, a slowly progressing Ab-mediated organ-specific autoimmune disease of the thyroid induced by immunization with syngeneic cells expressing TSHR. Flt3-L and GM-CSF treatment resulted in up-regulation of CD8a+ and CD8a− dendritic cells, and skewing of cytokine and immune responses to TSHR in favor of Th1 and Th2, respectively. However, this skewing did not persist until the later stages, and thus failed to affect the course or severity of the disease. To determine whether the total absence of either IL-4 or IFN-γ could affect the development of EAGD, we immunized wild-type, IFN-γ−/− and IL-4−/− BALB/c mice with TSHR. Nearly 100% of the wild-type and IFN-γ−/− mice developed EAGD with optimal TSHR-specific immune responses, while IL-4−/− mice completely resisted disease and showed delayed and suboptimal pathogenic Ab response. These data demonstrated that skewing immune responses to TSHR, using either Flt3-L or GM-CSF, in favor of Th1 or Th2, respectively, may not be sufficient to alter the course of the disease, while the complete absence of IL-4, but not IFN-γ, can prevent the development of EAGD.

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“…Purification has been hampered by the relatively small number of TSHR molecules expressed in mammalian cells and the functional instability of the protein. An important factor contributing to this difficulty is that TSHR autoantibodies predominantly recognize highly conformational epitopes (which are not necessarily identical with those that bind TSH) (Nagayama et al 1991, Tahara et al 1991.…”

Section: Introductionmentioning

confidence: 99%

Isolation of radiochemically pure 125I-labeled human thyrotropin receptor and its use for the detection of pathological autoantibodies in sera from Graves' patients

Loos²

1999

Journal of Endocrinology

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We report a method for the purification and radioactive labeling of human TSH receptor (TSHR). The method is based on the construction of a fusion TSHR (TSHR-Xa-BIO) which consists of the N-terminal 725 amino acids of human TSHR linked to the 4-amino acid Xa protease cleavage site and the 87-amino acid C-terminal domain of the biotin carboxyl carrier protein subunit of Escherichia coli acetyl-CoA carboxylase (the C-terminal domain directs the efficient posttranslational biotinylation of the protein). TSHR-Xa-BIO was produced in HeLa cells using recombinant vaccinia virus. The expressed protein was fully functional and was biotinylated with an efficiency of about 90%. Streptavidin-agarose-immobilized TSHRXa-BIO was labeled with 125 I using the chloramine T oxidation procedure and specifically eluted from the solid phase after cleavage with protease Xa. Isolated native radiochemically pure 125 I-labeled TSHR specifically interacted with pathological autoantibodies in the sera of patients with Graves' disease, and thus could be useful for the detection of these autoantibodies by immunoprecipitation analysis.

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Immunoglobulins from Graves' disease patients interact with different sites on TSH receptor/LH-CG receptor chimeras than either TSH or immunoglobulins from idiopathic myxedema patients

Cited by 135 publications

References 14 publications

A Novel Murine Model of Graves’ Hyperthyroidism with Intramuscular Injection of Adenovirus Expressing the Thyrotropin Receptor

A Novel Murine Model of Graves’ Hyperthyroidism with Intramuscular Injection of Adenovirus Expressing the Thyrotropin Receptor

Absence of IL-4, and Not Suppression of the Th2 Response, Prevents Development of Experimental Autoimmune Graves’ Disease

Isolation of radiochemically pure 125I-labeled human thyrotropin receptor and its use for the detection of pathological autoantibodies in sera from Graves' patients

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