Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jun;42(3):109-114.
doi: 10.1089/mab.2022.0037.

Novel Antimurine Thyroid-Stimulating Hormone Receptor Monoclonal Antibodies

Alyssa N Klee  1   2 , James A Torchia  1   2 , Gordon J Freeman  1   2
Affiliations

Novel Antimurine Thyroid-Stimulating Hormone Receptor Monoclonal Antibodies

Alyssa N Klee et al. Monoclon Antib Immunodiagn Immunother. 2023 Jun.

Abstract

Autoantibodies against thyroid proteins are present in several thyroid diseases. Thyroid-stimulating hormone receptor (TSHR) is a G-protein-coupled receptor (GPCR) that binds to thyroid-stimulating hormone (TSH) and stimulates production of thyroxine (T4) and triiodothyronine (T3). When agonized by anti-TSHR autoantibodies, aberrant production of thyroid hormone can lead to Graves' Disease (GD). In Hashimoto's thyroiditis (HT), anti-TSHR autoantibodies target the thyroid for immune attack. To better understand the role of anti-TSHR antibodies in thyroid disease, we generated a set of rat antimouse (m)TSHR monoclonal antibodies with a range of affinities, blocking of TSH, and agonist activity. These antibodies could be used to investigate the etiology and therapy of thyroid disease in mouse models and as building blocks in protein therapeutics that target the thyroid for treatment in either HT or GD.

Keywords: Graves disease; T3; T4; TSHR; monoclonal antibody; thyroid.

PubMed Disclaimer

Conflict of interest statement

G.J.F. has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. G.J.F. has served on advisory boards for Roche, Bristol-Myers-Squibb, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, GV20, IOME, and Geode. G.J.F. has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, and Geode.

Figures

FIG. 1.
FIG. 1.
TSH—TSHR—T3 pathway. TRH stimulates the anterior pituitary gland to produce TSH, which then stimulates TSHR. TSHR is a GPCR, which when stimulated activates the Gα and AC. AC generates cAMP to activate the transcription factor CREB. CREB is critical for T3 production. After T3 and T4 are produced, they travel through the circulation and are transported into the nuclei of peripheral tissue cells where they bind to TRs (thyroid hormone receptors) with its heterodimeric partner the RXR. This complex increases the transcription of genes with a TRE in their promoter. AC, adenyl cyclase; cAMP, cyclic adenosine monophosphate; CREB, cAMP response element-binding protein; Gα, alpha G subunit; GPCR, G-protein coupled receptor; RXR, retinoid X receptor; T3, triiodothyronine; T4, thyroxine; TRE, thyroid response element; TRH, thyrotropin releasing hormone; TSH, thyroid-stimulating hormone; TSHR, thyroid-stimulating hormone receptor.
FIG. 2.
FIG. 2.
Characterization of a panel of anti-mTSHR antibodies. 300.19 cells expressing murine TSHR were stained with a panel of antimurine TSHR mAbs as hybridoma supernatants followed by goat antirat IgG-PE. mAbs, monoclonal antibodies; MFI, mean fluorescence intensity.
FIG. 3.
FIG. 3.
Functional characterization of anti-TSHR antibodies. (A) Antibody that binds outside of the TSH binding site permits TSH binding to TSHR (left). Antibody that binds to the TSH binding site blocks TSH from binding to TSHR (right). (B) 300.19-TSHR cells incubated with the indicated antibodies, then stained with fluorescently labeled TSH-AF647. (C) 300.19-TSHR cells incubated with the indicated antibodies and then stained with fluorescently labeled TSH-AF647. (D) 300.19 cells expressing murine TSHR were stained with the three indicated purified antimurine TSHR mAbs followed by goat antirat IgG-PE. Purified rat IgG2b was used as an isotype control.
FIG. 4.
FIG. 4.
T3 production by C57BL/6 mice in response to intraperitoneal TSH injection. C57BL/6 mice were injected with PBS, TSH, or 1F8 antibody clone (five mice per group). Blood samples from 4 and 24 hours were assayed to quantify T3 levels.
See this image and copyright information in PMC

References

    1. Ihnatowicz P, Drywień M, Wątor P, et al. . The importance of nutritional factors and dietary management of Hashimoto's thyroiditis. Ann Agric Environ Med 2020;27:184–193. - PubMed
    1. Garber JR, Cobin RH, Gharib H, et al. . Clinical practice guidelines for hypothyroidism in adults: cosponsored by the American Association of Clinical Endocrinologists and the American Thyroid Association. Endocr Pract 2012;18:988–1028. - PubMed
    1. Mincer DL, Jialal I.. Hashimoto Thyroiditis. StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2022. [cited 2022 May 30]. Available from: http://www.ncbi.nlm.nih.gov/books/NBK459262 [Last accessed: May 30, 2022].
    1. Akamizu T, Amino N.. Hashimoto's thyroiditis. In: Endotext [Internet]. (Feingold KR, Anawalt B, Boyce A, Chrousos G, de Herder WW, Dhatariya K, et al. eds.) MDText.com, Inc.; South Dartmouth (MA); 2000 [cited 2022 May 31]. Available from: http://www.ncbi.nlm.nih.gov/books/NBK285557 [Last accessed: May 30, 2022].
    1. Morshed SA, Ma R, Latif R, et al. . Biased signaling by thyroid-stimulating hormone receptor–specific antibodies determines thyrocyte survival in autoimmunity. Sci Signal 2018;11:eaah4120. - PMC - PubMed

LinkOut - more resources