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Published on: September 1, 2015
Dendritic cells undergo rapid apoptosis in vitro during antigen-specific interaction with CD4+ T cells
H Matsue1, D Edelbaum, A C Hartmann
1Department of Dermatology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Insights
T cells induce dendritic cell (DC) apoptosis after antigen-specific interactions. This T cell-mediated DC death, involving CD95 (Fas) and CD95 ligand (CD95L), acts as a regulatory mechanism to prevent prolonged T cell activation.
Area of Science:
- Immunology
- Cell Biology
Background:
- The ultimate fate of dendritic cells (DCs) after interacting with T cells is not fully understood.
- Dendritic cells play a crucial role in initiating and regulating immune responses.
Purpose of the Study:
- To investigate whether dendritic cells undergo apoptosis following antigen-specific interactions with T cells.
- To elucidate the mechanisms involved in T cell-induced dendritic cell death.
Main Methods:
- Coculture of splenic dendritic cells and a T cell clone with a specific antigen.
- Assessment of cell death using propidium iodide uptake and chromatin condensation.
- Analysis of CD95 (Fas) and CD95 ligand (CD95L) expression using RT-PCR and FACS.
- Inhibition studies using anti-CD95L monoclonal antibody (mAb) and anti-CD95 mAb with cycloheximide.
- Evaluation of delayed type hypersensitivity responses in vivo using dendritic cells from CD95-deficient mice.
Main Results:
- Dendritic cells exhibited significant cell death when cocultured with antigen-specific T cells and antigen, but not with either alone.
- T cell-mediated dendritic cell death was partially inhibited by anti-CD95L mAb.
- Ligation of CD95 on dendritic cells with CD95L on T cells induced cell death, particularly with cycloheximide.
- Dendritic cells from CD95-deficient mice were more effective in inducing delayed type hypersensitivity responses in vivo.
Conclusions:
- T cell-induced dendritic cell apoptosis occurs after antigen-specific interactions.
- The CD95/CD95L pathway is involved in, but not solely responsible for, T cell-mediated dendritic cell death.
- T cell-induced dendritic cell apoptosis serves as a regulatory mechanism to limit T cell activation by antigen-presenting dendritic cells.
Abstract:
The terminal fate of dendritic cells (DC) remains relatively uncertain. In this study, we tested the hypothesis that DC undergo apoptosis after Ag-specific interaction with T cells. When splenic DC isolated from BALB/c mice were cocultured with HDK-1 T cells (a keyhole limpet hemocyanin (KLH)-specific CD4+ Th1 clone) in the presence of KLH, they showed conspicuous cell death as measured by propidium iodide (PI) uptake and chromatin condensation, whereas they remained relatively intact when incubated with either T cells or KLH alone. Likewise, the long term DC line XS52, which was established from BALB/c mouse epidermis, also died rapidly (within 2 h), and they exhibited characteristic DNA laddering when cocultured with HDK-1 T cells in the presence of KLH. RT-PCR and FACS analyses revealed the expression of CD95 (Fas) by XS52 DC and of CD95 ligand (CD95L) (Fas ligand) by activated HDK-1 T cells, suggesting a functional role for these molecules. In fact, anti-CD95L mAb inhibited partially (50%) T cell-mediated XS52 cell death, and coupling of surface CD95 with anti-CD95 mAb triggered significant XS52 cell death, but only in the presence of cycloheximide. Thus, ligation of CD95 (on DC) with CD95L (on T cells) is one, but not the only, mechanism by which T cells induce DC death. Finally, DC isolated from the CD95-deficient mice were found to be significantly more efficient than DC from control mice in their capacity to induce delayed type hypersensitivity responses in vivo. We propose that T cell-induced DC apoptosis serves as a unique down-regulatory mechanism that prevents the interminable activation of T cells by Ag-bearing DC.
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