title: Retention of cyclic AMP response to TSH in a cloned human thyrocyte/T cell hybridoma (HY2-15).
abstract: Our observation of a human T cell leukemia cell (Molt 4) demonstrating low affinity thyroid-stimulating hormone (TSH) responses, as evidenced by generation of cyclic AMP, led us to test Molt 4 cells as a suitable partner for immortalizing high affinity TSH receptors present on human thyroid cells. Therefore, we generated a hybridoma (HY2-15) by a fusion between thyroid monolayer cells from a patient with Graves' disease, and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive variant of this human T cell leukemia line, Molt 4-8AGR. The hybrid nature of HY2-15 was confirmed by DNA histograms using propidium iodide and flow cytometry. Karyotyping showed the HY2-15 cells to have five sets of chromosomes and human leukocyte antigen (HLA) class I determination revealed the presence of an additional HLA class I antigen (A2) not present on the Molt 4 partner cells. The established, cloned, hybridoma cells showed a greater than 30-fold increase in cyclic AMP release after stimulation with bovine TSH (bTSH, 1 mU/ml) with a minimum detectable stimulating dose of less than 10 microU/ml bTSH. However, no other thyroid-specific functions could be detected. Furthermore, HY2-15 cells failed to express HLA class II antigens either constitutively or in response to recombinant human gamma interferon (IF) and a variety of other stimuli, data similar to the Molt 4 partner cells but in contrast to human thyroid cells which show high sensitivity to gamma IF. The preservation of highly sensitive TSH responsiveness in a proliferating cell offers a unique approach to the study of human TSH receptor function.
authors.affiliation: Department of Medicine, Mount Sinai School of Medicine, New York, NY.
authors.forename: A
authors.initials: A
authors.lastname: Martin
authors.forename: M
authors.initials: M
authors.lastname: Platzer
authors.forename: T F
authors.initials: TF
authors.lastname: Davies
mesh_terms: D002455:Cell Division; D002999:Clone Cells; D000242:Cyclic AMP / Q000096:biosynthesis*; D004305:Dose-Response Relationship, Drug; D006111:Graves Disease; D006680:HLA Antigens / Q000032:analysis / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007371:Interferon-gamma / Q000494:pharmacology; D007621:Karyotyping; D015458:Leukemia, T-Cell / Q000378:metabolism*; D010641:Phenotype; D011989:Receptors, Thyrotropin / Q000502:physiology; D011994:Recombinant Proteins; D013972:Thyrotropin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D006680:HLA Antigens; D011989:Receptors, Thyrotropin; D011994:Recombinant Proteins; D007371:Interferon-gamma; D013972:Thyrotropin; D000242:Cyclic AMP