HLA-DR gene expression in a proliferating human thyroid cell clone (12S).
We have used a retroviral vector carrying the adenovirus E1A oncogene and the neomycin phosphotransferase gene to establish a human thyroid-derived cell line that exhibits TSH-mediated cAMP generation as well as the differential expression of HLA class II antigens in response to recombinant gamma-interferon. Twenty-two-week gestation, histologically confirmed, human fetal thyroid was collagenase digested, cultured as a monolayer, and infected directly with 12S or 13S E1A-containing retrovirus constructs. Infected clones (n = 30) were selected in a hormone-supplemented medium containing bovine TSH (bTSH; 1 mU/ml), 10% fetal bovine serum, and 0.5 mg/ml G418 antibiotic. A rapidly growing clone (designated 12S) was chosen for detailed analysis over 18 months of continuous culture. The 12S clone was sensitive to less than 10 microU/ml bTSH when assessed by extracellular accumulation of cAMP, but TSH had no influence on 72-h incorporation of [3H]thymidine. Clone 12S responded to recombinant human gamma-interferon (1-10(4) U/ml) by induction of HLA DR alpha-chain-specific mRNA and the surface expression of HLA-DR antigen detected by fluorescein isothiocyanate-labeled monoclonal antibody to nonpolymorphic HLA-DR regions using flow cytometry. These studies indicate the potential for immortalizing human thyroid cells for use as targets of anti-TSH receptor immune responses and for long term studies of human throcyte HLA gene regulation.
Cone
R D
RD
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029.
Platzer
M
M
Piccinini
L A
LA
Jaramillo
M
M
Davies
T F
TF
D000328:Adult; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005333:Fetus; D005802:Genes, MHC Class II; D005822:Genetic Vectors; D006681:HLA-D Antigens / Q000235:genetics*; D006684:HLA-DR Antigens / Q000235:genetics*; D006801:Humans; D008285:Major Histocompatibility Complex / Q000187:drug effects; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D013972:Thyrotropin / Q000494:pharmacology; D014158:Transcription, Genetic
D006681:HLA-D Antigens; D006684:HLA-DR Antigens; D013972:Thyrotropin; D004262:DNA Restriction Enzymes