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Use of cells expressing the human thyrotropin (TSH) receptor for the measurement of thyroid stimulating and TSH-blocking antibodies.
Acta Med Austriaca 1996; 23(1-2):52-6AM

Abstract

A clone of Chinese hamster ovary cells (CHO) transfected with the cDNA of the human thyrotropin (TSH) receptor (CHO-R) was used to optimise assays for TSH receptor antibodies with either thyroid stimulating (TSAb) or TSH blocking (TSH-blocking Ab) activity. The study group included 89 patients with Graves' disease, 38 patients with goitrous Hashimoto's thyroiditis (HT) and 47 subjects with atrophic thyroiditis (AT). In the HT group, 8 patients had subclinical hypothyroidism (HT-SH) and 30 had overt hypothyroidism (HT-H). In the assay for TSAb, CHO-R cells were incubated with 1mg/ml of the immunoglobulin G (IgG) from patients with Grave's disease, while in the assay for TSH-blocking Ab cells were incubated with IgGs from patients with HT or AT alone (1mg/ml), or IgGs plus TSH (10 mU/L). After 2 h of incubation the extracellular cAMP was measured by a RIA. In these conditions a significant stimulation by Graves' IgG was obtained in patients with active hyperthyroidism (33/35, 94% untreated; 21/23, 91% relapsed after a course of medical treatment), in 12/20 (60%) patients euthyroid under methimazole and in 4/11 (36%) euthyroid after a course of antithyroid drugs. TSH-blocking Ab were detected in 1/8 (12.5%) patients with HT-SH, in 7/30 (23.3%) with HT-H and in 16/47 (34.0%) patients with AT. TSAb and TSH-blocking Ab coexisted in 4 IgGs that belonged to patients in whom spontaneous hypothyroidism developed after hyperthyroidism, or viceversa. TSAb and TSH-blocking Ab were also tested on FTRL-5 cells. TSAb were positive in both assays in 43/58 (74%) patients with active Graves' disease, negative in both assays in 3 (5%), negative in FRTL-5 and positive in CHO-R in 11 (19%), negative in CHO-R and positive in FRTL-5 in 1 (1.7%). In FTRL-5 cells TSH-blocking Ab were detected in 1/8 (12.5%) patients with HT-SH, in 5/30 (16.6%) with HT-H and in 15/47 (31.9%) with AT. The results of cAMP stimulation in FRTL-5 and CHO-R showed a fairly good correlation in TSAb (r = 0.60, p < 0.0001) and in TSH-blocking Ab (r = 0.74, p < 0.0001) assays. In conclusion, CHO cells transfected with the cloned human TSH receptor are suitable for the clinical assay of TSAb and TSH-blocking Ab. The sensitivity of this assay is higher than that obtained using FRTL-5 cells, having the additional advantages of expressing the human TSH receptor and requiring less cumbersome procedures for cell culture.

Authors+Show Affiliations

Institute of Endocrinology, University of Pisa, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

8767515

Citation

Vitti, P, et al. "Use of Cells Expressing the Human Thyrotropin (TSH) Receptor for the Measurement of Thyroid Stimulating and TSH-blocking Antibodies." Acta Medica Austriaca, vol. 23, no. 1-2, 1996, pp. 52-6.
Vitti P, Chiovato L, Fiore E, et al. Use of cells expressing the human thyrotropin (TSH) receptor for the measurement of thyroid stimulating and TSH-blocking antibodies. Acta Med Austriaca. 1996;23(1-2):52-6.
Vitti, P., Chiovato, L., Fiore, E., Mammoli, C., Rocchi, R., & Pinchera, A. (1996). Use of cells expressing the human thyrotropin (TSH) receptor for the measurement of thyroid stimulating and TSH-blocking antibodies. Acta Medica Austriaca, 23(1-2), pp. 52-6.
Vitti P, et al. Use of Cells Expressing the Human Thyrotropin (TSH) Receptor for the Measurement of Thyroid Stimulating and TSH-blocking Antibodies. Acta Med Austriaca. 1996;23(1-2):52-6. PubMed PMID: 8767515.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Use of cells expressing the human thyrotropin (TSH) receptor for the measurement of thyroid stimulating and TSH-blocking antibodies. AU - Vitti,P, AU - Chiovato,L, AU - Fiore,E, AU - Mammoli,C, AU - Rocchi,R, AU - Pinchera,A, PY - 1996/1/1/pubmed PY - 1996/1/1/medline PY - 1996/1/1/entrez SP - 52 EP - 6 JF - Acta medica Austriaca JO - Acta Med. Austriaca VL - 23 IS - 1-2 N2 - A clone of Chinese hamster ovary cells (CHO) transfected with the cDNA of the human thyrotropin (TSH) receptor (CHO-R) was used to optimise assays for TSH receptor antibodies with either thyroid stimulating (TSAb) or TSH blocking (TSH-blocking Ab) activity. The study group included 89 patients with Graves' disease, 38 patients with goitrous Hashimoto's thyroiditis (HT) and 47 subjects with atrophic thyroiditis (AT). In the HT group, 8 patients had subclinical hypothyroidism (HT-SH) and 30 had overt hypothyroidism (HT-H). In the assay for TSAb, CHO-R cells were incubated with 1mg/ml of the immunoglobulin G (IgG) from patients with Grave's disease, while in the assay for TSH-blocking Ab cells were incubated with IgGs from patients with HT or AT alone (1mg/ml), or IgGs plus TSH (10 mU/L). After 2 h of incubation the extracellular cAMP was measured by a RIA. In these conditions a significant stimulation by Graves' IgG was obtained in patients with active hyperthyroidism (33/35, 94% untreated; 21/23, 91% relapsed after a course of medical treatment), in 12/20 (60%) patients euthyroid under methimazole and in 4/11 (36%) euthyroid after a course of antithyroid drugs. TSH-blocking Ab were detected in 1/8 (12.5%) patients with HT-SH, in 7/30 (23.3%) with HT-H and in 16/47 (34.0%) patients with AT. TSAb and TSH-blocking Ab coexisted in 4 IgGs that belonged to patients in whom spontaneous hypothyroidism developed after hyperthyroidism, or viceversa. TSAb and TSH-blocking Ab were also tested on FTRL-5 cells. TSAb were positive in both assays in 43/58 (74%) patients with active Graves' disease, negative in both assays in 3 (5%), negative in FRTL-5 and positive in CHO-R in 11 (19%), negative in CHO-R and positive in FRTL-5 in 1 (1.7%). In FTRL-5 cells TSH-blocking Ab were detected in 1/8 (12.5%) patients with HT-SH, in 5/30 (16.6%) with HT-H and in 15/47 (31.9%) with AT. The results of cAMP stimulation in FRTL-5 and CHO-R showed a fairly good correlation in TSAb (r = 0.60, p < 0.0001) and in TSH-blocking Ab (r = 0.74, p < 0.0001) assays. In conclusion, CHO cells transfected with the cloned human TSH receptor are suitable for the clinical assay of TSAb and TSH-blocking Ab. The sensitivity of this assay is higher than that obtained using FRTL-5 cells, having the additional advantages of expressing the human TSH receptor and requiring less cumbersome procedures for cell culture. SN - 0303-8173 UR - https://www.unboundmedicine.com/medline/citation/8767515/Use_of_cells_expressing_the_human_thyrotropin__TSH__receptor_for_the_measurement_of_thyroid_stimulating_and_TSH_blocking_antibodies_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&amp;sid=nlm:pubmed&amp;issn=0303-8173&amp;date=1996&amp;volume=23&amp;issue=1-2&amp;spage=52 DB - PRIME DP - Unbound Medicine ER -