In vitro maturation of clonal CD4+CD8+ cell lines in response to TCR engagement.
Engagement of the TCR on immature CD4+CD8+ (DP) thymocytes by an appropriate peptide/MHC ligand evokes a complex program of maturation known as positive selection. As a result, DP thymocytes are rescued from programmed cell death, become committed to the CD4 or CD8 lineage, extinguish expression of V(D)J recombinase activity, and undergo further maturation. We describe here a panel of DP thymic lymphoma cell lines that, in response to in vitro TCR engagement, undergo many of the TCR-beta-induced maturation events that have been reported to accompany positive selection of DP thymocytes in vivo. These events include increased expression of CD5, CD69, CD45, TCR-alpha, and MHC class I, and decreased expression of Thy-1 and heat-stable Ag. In addition, we observed TCR-induced expression of the bcl-2 gene, a well described inhibitor of programmed cell death. Finally, TCR engagement decreased expression of recombinase-activating genes and terminal deoxynucleotidal transferase genes, as well as V(D)J recombinase activity. However, TCR engagement did not elicit demonstrable CD4/CD8 lineage commitment. These observations suggest that engagement of the TCR on these DP cell lines elicits multiple maturation events that are part of the positive selection developmental program, but not CD4/CD8 lineage commitment. Thus, these DP cell lines provide the opportunity to elucidate molecular mechanisms of maturation and CD4/CD8 lineage commitment in vitro.
Groves
T
T
Division of Immunology and Cancer, Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
Katis
P
P
Madden
Z
Z
Manickam
K
K
Ramsden
D
D
Wu
G
G
Guidos
C J
CJ
D000818:Animals; D017209:Apoptosis / Q000276:immunology; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000276:immunology*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D005434:Flow Cytometry; D016130:Immunophenotyping; D008213:Lymphocyte Activation / Q000276:immunology; D008223:Lymphoma / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D011948:Receptors, Antigen, T-Cell / Q000276:immunology* / Q000378:metabolism*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000502:physiology*; D013950:Thymus Gland / Q000166:cytology; D013953:Thymus Neoplasms / Q000276:immunology; D014407:Tumor Cells, Cultured; D045567:VDJ Recombinases
D011948:Receptors, Antigen, T-Cell; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases