Studies on the cyclic AMP response to thyroid stimulating immunoglobulin (TSI) and thyrotropin (TSH) in human thyroid cell monolayers.
Studies were conducted on the cultured human thyroid cell bioassay for thyroid stimulating immunoglobulin (TSI) and thyrotropin (TSH). In confirmation of the data of Kasagi, et al.,15 incubation of human thyroid cells in Hank's balanced salt solution deficient in NaCl increased the sensitivity of the c-AMP response to bovine TSH by approximately one order of magnitude. Half-maximal stimulation was attained at approximately 0.1 mU bTSH/ml. The effect of NaCl in the medium was greater with stimulation by TSI greater than hTSH greater than bTSH. In contract to incubations in NaCl(+)medium, with NaCl(-) medium most (70%-80%) of the c-AMP produced was released into the medium; this proportion remaining relatively constant over a wide range of bTSH and hTSH concentrations. At TSI concentrations higher than 3mg/ml efflux of c-AMP into the medium was significantly diminished. Stimulation by cholera toxin and prostaglandin E of thyroid cell c-AMP generation was not enhanced in NaCl(-) medium, in contrast to stimulation by TSH and TSI. The presence of 10(-4)m cycloheximide in NaCl(+) medium enhanced the c-AMP response to physiological concentrations of TSH. As with NaCl(-) medium, cycloheximide increased the sensitivity but not the maximum response of the c-AMP response to TSH. However no additivity was observed with NaCl(-) medium and cycloheximide. Human thyroid cells obtained from patients with Graves' disease are relatively insensitive to TSI stimulation. In NaCl(-) medium, however, the sensitivity of these cells to TSI stimulation is sufficient to enable them to be utilized in the TSI assay. The present state of the TSI assay is discussed.
D000527:Alprostadil; D000818:Animals; D001681:Biological Assay; D002417:Cattle; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis* / Q000494:pharmacology; D018828:Immunoglobulins, Thyroid-Stimulating; D011458:Prostaglandins E / Q000494:pharmacology; D014176:Protein Biosynthesis; D012965:Sodium Chloride / Q000494:pharmacology; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*; D013972:Thyrotropin / Q000032:analysis* / Q000494:pharmacology
D007074:Immunoglobulin G; D018828:Immunoglobulins, Thyroid-Stimulating; D011458:Prostaglandins E; D012965:Sodium Chloride; D013972:Thyrotropin; D002772:Cholera Toxin; D000242:Cyclic AMP; D000527:Alprostadil