De novo HLA class II and enhanced HLA class I molecule expression in SV40 transfected human thyroid epithelial cells.
Human thyroid follicular cells normally synthesize and express HLA Class I molecules but in glands affected by autoimmune or neoplastic diseases they express Class II molecules. We have investigated the effect of SV40 virus transformation on the expression of MHC molecules in human thyrocytes. Primary cultures of human thyroid from two different glands were transfected with the plasmid pX-8, containing the 'early' region of SV40 virus, and two continuous lines of thyrocytes were obtained. The cell lines maintained features of parental thyroid epithelial cells showing the characteristic cytokeratin filament network, microvillar protrusion and tight junctions. In addition, the SV40-transfected cells responded to graded doses of TSH with increased c-AMP production. Thyrocytes from both cell lines hyperexpressed Class I molecules and a significant proportion of them also acquired constitutive Class II expression, as determined by indirect immunofluorescence (IFL), flow cytometry and Northern blotting hybridization using a DR beta probe. These cells were found to be normally up-regulated by interferon (IFN)-gamma. Indirect IFL and flow cytometry analysis were used to detect and quantify the expression of HLA-DR, DP and DQ subregions. A co-ordinated expression (DR greater than DP much greater than DQ), reminiscent of the inappropriate HLA expression found in thyroid autoimmune disease in vivo and of the in vitro regulation in normal thyrocytes, was observed. Clones derived from these cell lines differed in their level of constitutive Class II expression and in their sensitivity to Class II induction by IFN-gamma. In conclusion, these thyroid cell lines could provide a useful tool for further investigation of HLA gene regulation in thyroid cells and for elucidating on the mechanism involved in the inappropriate HLA expression described in autoimmune and neoplastic diseases of the thyroid.
Belfiore
A
A
Department of Immunology, University College and Middlesex School of Medicine, London, UK.
Mauerhoff
T
T
Pujol-Borrell
R
R
Badenhoop
K
K
Buscema
M
M
Mirakian
R
R
Bottazzo
G F
GF
D001324:Autoantigens / Q000378:metabolism; D002461:Cell Line, Transformed; D000242:Cyclic AMP / Q000378:metabolism; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology / Q000378:metabolism; D006681:HLA-D Antigens / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D007453:Iodide Peroxidase; D033862:Iron-Binding Proteins; D013539:Simian virus 40 / Q000235:genetics; D013954:Thyroglobulin / Q000378:metabolism; D013961:Thyroid Gland / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013972:Thyrotropin / Q000494:pharmacology; D014162:Transfection
D001324:Autoantigens; D006681:HLA-D Antigens; D015395:Histocompatibility Antigens Class I; D033862:Iron-Binding Proteins; D013972:Thyrotropin; D013954:Thyroglobulin; D000242:Cyclic AMP; C056658:TPO protein, human; D007453:Iodide Peroxidase