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Induction of interleukin 10-producing, nonproliferating CD4(+) T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells.

Jonuleit H, Schmitt E, Schuler G, Knop J, Enk AH - J. Exp. Med. (2000)

Bottom Line: The functional properties of dendritic cells (DCs) are strictly dependent on their maturational state.Furthermore, in coculture experiments these T cells inhibited the antigen-driven proliferation of Th1 cells in a contact- and dose-dependent, but antigen-nonspecific manner.These data show that immature and mature DCs induce different types of T cell responses: inflammatory Th1 cells are induced by mature DCs, and IL-10-producing T cell regulatory 1-like cells by immature DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Mainz, D-55101 Mainz, Germany. jonuleit@hautklinik.klinik.uni-mainz.de

ABSTRACT
The functional properties of dendritic cells (DCs) are strictly dependent on their maturational state. To analyze the influence of the maturational state of DCs on priming and differentiation of T cells, immature CD83(-) and mature CD83(+) human DCs were used for stimulation of naive, allogeneic CD4(+) T cells. Repetitive stimulation with mature DCs resulted in a strong expansion of alloreactive T cells and the exclusive development of T helper type 1 (Th1) cells. In contrast, after repetitive stimulation with immature DCs the alloreactive T cells showed an irreversibly inhibited proliferation that could not be restored by restimulation with mature DCs or peripheral blood mononuclear cells, or by the addition of interleukin (IL)-2. Only stimulation of T cells with mature DCs resulted in an upregulation of CD154, CD69, and CD70, whereas T cells activated with immature DCs showed an early upregulation of the negative regulator cytotoxic T lymphocyte-associated molecule 4 (CTLA-4). These T cells lost their ability to produce interferon gamma, IL-2, or IL-4 after several stimulations with immature DCs and differentiated into nonproliferating, IL-10-producing T cells. Furthermore, in coculture experiments these T cells inhibited the antigen-driven proliferation of Th1 cells in a contact- and dose-dependent, but antigen-nonspecific manner. These data show that immature and mature DCs induce different types of T cell responses: inflammatory Th1 cells are induced by mature DCs, and IL-10-producing T cell regulatory 1-like cells by immature DCs.

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Inhibition of antigen-specific proliferation of Th1 cells after coculture with Tr1-like cells. Th1 and Tr1 cell lines were induced by repetitive stimulation of naive CD4+ T cells with allogeneic mDCs (Th1 cytokine profile) or iDCs (Tr1 cytokine profile) at DC/T cell ratios of 1:10. 7 d after the third restimulation, the Th1 cells (5 × 104 cells/well) were restimulated with mDCs (5 × 103 cells/well) in the presence of different numbers of Tr1-like cells from the same cord blood fraction induced with iDCs from the same DC donor. Proliferation of T cells was determined by [3H]TdR incorporation after 4 d of culture. *Background proliferation of Tr1-like cells plus mDCs. One of three experiments is shown.
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Figure 6: Inhibition of antigen-specific proliferation of Th1 cells after coculture with Tr1-like cells. Th1 and Tr1 cell lines were induced by repetitive stimulation of naive CD4+ T cells with allogeneic mDCs (Th1 cytokine profile) or iDCs (Tr1 cytokine profile) at DC/T cell ratios of 1:10. 7 d after the third restimulation, the Th1 cells (5 × 104 cells/well) were restimulated with mDCs (5 × 103 cells/well) in the presence of different numbers of Tr1-like cells from the same cord blood fraction induced with iDCs from the same DC donor. Proliferation of T cells was determined by [3H]TdR incorporation after 4 d of culture. *Background proliferation of Tr1-like cells plus mDCs. One of three experiments is shown.

Mentions: To analyze the functional properties of IL-10–producing Tr1-like cells, coculture experiments with alloreactive Th1 cell lines were performed. To this end, alloreactive Th1 cell lines were restimulated with mDCs in the presence of different numbers of Tr1-like cells (also stimulated with mDCs) from the same donor. As shown in Fig. 6, Tr1-like cells suppressed the proliferation of syngeneic Th1 cells in response to allogeneic mDCs in a dose-dependent manner. At a Th1/Tr1 ratio of 1:1, the proliferation of Th1 cells is reduced to nearly the level of proliferation of the activated Tr1-like cells alone (Tr1-like cells plus mDCs, ratio of 1:10; Fig. 6). The observation that activated Tr1-like cells produce high levels of the immunosuppressive cytokine IL-10 and no T cell growth–promoting cytokines such as IL-2 and IL-4 suggests that the inhibitory effect of Tr1-like cells on the proliferation of Th1 cells might be a direct result of soluble inhibitory factors produced by activated Tr1-like cells. To test this hypothesis, alloreactive Th1 and Tr1-like cells were stimulated with allogeneic iDCs or mDCs. Additionally, alloreactive Tr1-like cells were either added directly to cocultures of activated Th1 cells or placed in Transwell chambers in the same well and activated with mDCs. The semipermeable polycarbonate membrane allows the free exchange of soluble factors but excludes direct cell contact of Th1 and Tr1-like cells. As shown in Fig. 7, alloreactive Th1 cell lines proliferated in the presence of allogeneic mDCs as well as in the presence of allogeneic iDCs. In contrast, Tr1-like cells with low proliferative capacity showed only weak rates of expansion, independent of the antigen-presenting population used. Furthermore, in coculture, Tr1-like cells suppressed the proliferation of Th1 cells almost to the level of Tr1-like cells induced by mDCs. This suggests that the proliferation of the cocultured Th1 cells is almost completely blocked. The separation of both T cell lines in Transwell chambers virtually abolished this immunosuppressive effect of Tr1-like cells. These results suggest that direct cell contact of Tr1 and Th1 cells is essential for the inhibitory capacity of Tr1-like cells. The comparatively low counts in Transwell experiments resulted from the experimental procedure (preculture in 24-well Transwell plates and transfer to 96-well plates at day 4) and is not a result of weak stimulatory capacity of mDCs in these assays.


Induction of interleukin 10-producing, nonproliferating CD4(+) T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells.

Jonuleit H, Schmitt E, Schuler G, Knop J, Enk AH - J. Exp. Med. (2000)

Inhibition of antigen-specific proliferation of Th1 cells after coculture with Tr1-like cells. Th1 and Tr1 cell lines were induced by repetitive stimulation of naive CD4+ T cells with allogeneic mDCs (Th1 cytokine profile) or iDCs (Tr1 cytokine profile) at DC/T cell ratios of 1:10. 7 d after the third restimulation, the Th1 cells (5 × 104 cells/well) were restimulated with mDCs (5 × 103 cells/well) in the presence of different numbers of Tr1-like cells from the same cord blood fraction induced with iDCs from the same DC donor. Proliferation of T cells was determined by [3H]TdR incorporation after 4 d of culture. *Background proliferation of Tr1-like cells plus mDCs. One of three experiments is shown.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2193357&req=5

Figure 6: Inhibition of antigen-specific proliferation of Th1 cells after coculture with Tr1-like cells. Th1 and Tr1 cell lines were induced by repetitive stimulation of naive CD4+ T cells with allogeneic mDCs (Th1 cytokine profile) or iDCs (Tr1 cytokine profile) at DC/T cell ratios of 1:10. 7 d after the third restimulation, the Th1 cells (5 × 104 cells/well) were restimulated with mDCs (5 × 103 cells/well) in the presence of different numbers of Tr1-like cells from the same cord blood fraction induced with iDCs from the same DC donor. Proliferation of T cells was determined by [3H]TdR incorporation after 4 d of culture. *Background proliferation of Tr1-like cells plus mDCs. One of three experiments is shown.
Mentions: To analyze the functional properties of IL-10–producing Tr1-like cells, coculture experiments with alloreactive Th1 cell lines were performed. To this end, alloreactive Th1 cell lines were restimulated with mDCs in the presence of different numbers of Tr1-like cells (also stimulated with mDCs) from the same donor. As shown in Fig. 6, Tr1-like cells suppressed the proliferation of syngeneic Th1 cells in response to allogeneic mDCs in a dose-dependent manner. At a Th1/Tr1 ratio of 1:1, the proliferation of Th1 cells is reduced to nearly the level of proliferation of the activated Tr1-like cells alone (Tr1-like cells plus mDCs, ratio of 1:10; Fig. 6). The observation that activated Tr1-like cells produce high levels of the immunosuppressive cytokine IL-10 and no T cell growth–promoting cytokines such as IL-2 and IL-4 suggests that the inhibitory effect of Tr1-like cells on the proliferation of Th1 cells might be a direct result of soluble inhibitory factors produced by activated Tr1-like cells. To test this hypothesis, alloreactive Th1 and Tr1-like cells were stimulated with allogeneic iDCs or mDCs. Additionally, alloreactive Tr1-like cells were either added directly to cocultures of activated Th1 cells or placed in Transwell chambers in the same well and activated with mDCs. The semipermeable polycarbonate membrane allows the free exchange of soluble factors but excludes direct cell contact of Th1 and Tr1-like cells. As shown in Fig. 7, alloreactive Th1 cell lines proliferated in the presence of allogeneic mDCs as well as in the presence of allogeneic iDCs. In contrast, Tr1-like cells with low proliferative capacity showed only weak rates of expansion, independent of the antigen-presenting population used. Furthermore, in coculture, Tr1-like cells suppressed the proliferation of Th1 cells almost to the level of Tr1-like cells induced by mDCs. This suggests that the proliferation of the cocultured Th1 cells is almost completely blocked. The separation of both T cell lines in Transwell chambers virtually abolished this immunosuppressive effect of Tr1-like cells. These results suggest that direct cell contact of Tr1 and Th1 cells is essential for the inhibitory capacity of Tr1-like cells. The comparatively low counts in Transwell experiments resulted from the experimental procedure (preculture in 24-well Transwell plates and transfer to 96-well plates at day 4) and is not a result of weak stimulatory capacity of mDCs in these assays.

Bottom Line: The functional properties of dendritic cells (DCs) are strictly dependent on their maturational state.Furthermore, in coculture experiments these T cells inhibited the antigen-driven proliferation of Th1 cells in a contact- and dose-dependent, but antigen-nonspecific manner.These data show that immature and mature DCs induce different types of T cell responses: inflammatory Th1 cells are induced by mature DCs, and IL-10-producing T cell regulatory 1-like cells by immature DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Mainz, D-55101 Mainz, Germany. jonuleit@hautklinik.klinik.uni-mainz.de

ABSTRACT
The functional properties of dendritic cells (DCs) are strictly dependent on their maturational state. To analyze the influence of the maturational state of DCs on priming and differentiation of T cells, immature CD83(-) and mature CD83(+) human DCs were used for stimulation of naive, allogeneic CD4(+) T cells. Repetitive stimulation with mature DCs resulted in a strong expansion of alloreactive T cells and the exclusive development of T helper type 1 (Th1) cells. In contrast, after repetitive stimulation with immature DCs the alloreactive T cells showed an irreversibly inhibited proliferation that could not be restored by restimulation with mature DCs or peripheral blood mononuclear cells, or by the addition of interleukin (IL)-2. Only stimulation of T cells with mature DCs resulted in an upregulation of CD154, CD69, and CD70, whereas T cells activated with immature DCs showed an early upregulation of the negative regulator cytotoxic T lymphocyte-associated molecule 4 (CTLA-4). These T cells lost their ability to produce interferon gamma, IL-2, or IL-4 after several stimulations with immature DCs and differentiated into nonproliferating, IL-10-producing T cells. Furthermore, in coculture experiments these T cells inhibited the antigen-driven proliferation of Th1 cells in a contact- and dose-dependent, but antigen-nonspecific manner. These data show that immature and mature DCs induce different types of T cell responses: inflammatory Th1 cells are induced by mature DCs, and IL-10-producing T cell regulatory 1-like cells by immature DCs.

Show MeSH
Related in: MedlinePlus