Establishment of a human thymic myoid cell line. Phenotypic and functional characteristics.
The subset of myoid cells is a normal component of the thymic stroma. To characterize these cells, we immortalized stromal cells from human thymus by using a plasmid vector encoding the SV40 T oncogene. Among the eight cell lines obtained, one had myoid characteristics including desmin and troponin antigens. This new line was designated MITC (myoid immortalized thymic cells). These cells expressed both the fetal and adult forms of muscle acetylcholine receptor (AChR) at the mRNA level, as well as the myogenic transcription factor MyoD1. alpha-Subunit AChR protein expression was detected by flow cytometry and the AChR was functional in patch-clamp studies. In addition, AChR expression was down-modulated by myasthenia gravis sera or by monoclonal antibody anti-AChR on MITC line similarly to TE671 rhabdomyosarcoma cells, making the MITC line an interesting tool for AChR antigenic modulation experiments. Finally, the MITC line expressed LFA-3, produced several cytokines able to act on T cells, and protected total thymocytes from spontaneous apoptosis in vitro. These results are compatible with a role of thymic myoid cells in some steps of thymocyte development. Therefore MITC line appears to be a useful tool to investigate the physiological role of thymic myoid cells.
Wakkach
A
A
CNRS ESA 8078, Hôpital Marie Lannelongue, Le Plessis-Robinsin, France Hôpital St-Antoine, Paris, France.
Poea
S
S
Chastre
E
E
Gespach
C
C
Lecerf
F
F
De La Porte
S
S
Tzartos
S
S
Coulombe
A
A
Berrih-Aknin
S
S
D016525:Antigenic Modulation / Q000187:drug effects; D000957:Antigens, Viral, Tumor / Q000096:biosynthesis / Q000235:genetics; D017209:Apoptosis; D001323:Autoantibodies / Q000494:pharmacology; D015152:Blotting, Northern; D002460:Cell Line; D002648:Child; D016207:Cytokines / Q000096:biosynthesis; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D066298:In Vitro Techniques; D009157:Myasthenia Gravis / Q000276:immunology; D017570:MyoD Protein / Q000096:biosynthesis; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D013950:Thymus Gland / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D014162:Transfection
D000957:Antigens, Viral, Tumor; D001323:Autoantibodies; D016207:Cytokines; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D012333:RNA, Messenger; D011950:Receptors, Cholinergic