Establishment of epithelial cell lines from human and mouse thymus immortalized by the 12S adenoviral E1a gene product.
To understand the role of thymic epithelial cells in the development of immature thymocytes, the establishment and cloning of thymic epithelial cell lines must be required. In the present study, we established human and mouse thymic epithelial cell lines through the immortalization by the transfection of cDNA sequences of adenoviral E1a 12S mRNA. This procedure resulted in the isolation of five stable cell lines (one human cell line and four mouse cell lines). These cell lines were positive in cytokeratin demonstrated by immunohistochemistry. Electron microscopic study revealed that they had bundles of tonofilaments and desmosome-like tight junctions. These findings indicate that the cell lines immortalized by E1a gene have retained the properties of epithelial cells. MHC class II antigens were not expressed on these cell lines. When interferon-gamma was added to the cultured epithelial cell lines, MHC class II antigens were induced in their cytoplasm and on their surface membrane, demonstrated by immunohistochemical and immunofluorescent studies. It is suggested that these stable cell lines from human and mouse thymus might serve a good tool for the further study of thymocytes differentiation and of unknown cytokines derived from thymus epithelium.
Hibi
T
T
Department of Internal Medicine, School of Medicine, Keio University.
Fujisawa
T
T
Kanai
T
T
Akatsuka
A
A
Habu
S
S
Handa
H
H
Tsuchiya
M
M
D000818:Animals; D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed / Q000502:physiology* / Q000648:ultrastructure; D002648:Child; D000229:Dependovirus; D004847:Epithelial Cells; D005802:Genes, MHC Class II; D005822:Genetic Vectors; D006801:Humans; D007633:Keratins / Q000032:analysis; D051379:Mice; D013950:Thymus Gland / Q000166:cytology*; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
D014764:Viral Proteins; D007633:Keratins