CD40 expressed on thymic epithelial cells provides costimulation for proliferation but not for apoptosis of human thymocytes.
Human thymic epithelial cells express CD40, so we examined the possible role of CD40 in activation of thymocytes. We observed that both CD4+CD8- and CD4-CD8+ thymocytes proliferate after stimulation by anti-CD3 mAb in the presence of cultured thymic epithelial cells. Costimulation of CD4+ thymocytes by thymic epithelial cells is partly inhibited by an anti-CD40 mAb, but this mAb has no effect on costimulation of CD8+ thymocytes. The selective costimulatory ability of CD40 for CD4+ thymocytes was confirmed in experiments in which thymocytes were stimulated with anti-CD3 in the presence of murine P815 cells transfected with CD40 cDNA. The level of costimulation induced by P815-CD40 was comparable with that induced by P815 cells expressing CD80 (B7.1). Treatment of thymocytes with the Ca2+ ionophore ionomycin and the phorbol ester PMA or with anti-CD3 mAb resulted in up-regulation of the CD40 ligand, suggesting that this molecule is involved in CD40-mediated costimulation of human thymocytes. Costimulation of thymocytes by CD80 strongly increased anti-CD3-induced death of fetal thymocytes. In contrast, costimulation by CD40 did not increase anti-CD3-mediated apoptosis of these thymocytes. To confirm that CD40 does not affect anti-CD3-induced cell death, we established a variant of the Jurkat T leukemic cell line that constitutively expresses CD40L and analyzed the sensitivity of this cell line for activation-induced apoptosis. In contrast to CD80, CD40 failed to increase anti-CD3-mediated apoptosis in CD40L+ Jurkat cells, whereas both CD40 and CD80 strongly increased IL-2 production induced by anti-CD3. These findings suggest that costimulation by CD40 is involved in clonal expansion of CD4+ thymocytes but not in activation-induced cell death.
Ruggiero
G
G
Division of Immunology of the Netherlands Cancer Institute, Amsterdam.
Martinez Cáceres
E
E
Voordouw
A
A
Noteboom
E
E
Graf
D
D
Kroczek
R A
RA
Spits
H
H
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis / Q000276:immunology*; D018122:B7-1 Antigen / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology; D019013:CD40 Antigens / Q000276:immunology / Q000378:metabolism* / Q000502:physiology*; D023201:CD40 Ligand; D016923:Cell Death / Q000276:immunology; D002675:Child, Preschool; D004848:Epithelium / Q000276:immunology / Q000378:metabolism; D005333:Fetus; D006801:Humans; D007223:Infant; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D015458:Leukemia, T-Cell; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*; D013950:Thymus Gland / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D018122:B7-1 Antigen; D017252:CD3 Complex; D019013:CD40 Antigens; D008562:Membrane Glycoproteins; D018799:Intercellular Adhesion Molecule-1; D023201:CD40 Ligand