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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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Related in: MedlinePlus

Proliferation of alloreactive CD4+ T cells induced by iDCs or mDCs. Naive CD4+ T cells purified by MACS sorting from cord blood were primed and restimulated (every week) with allogeneic iDCs or mDCs from the same donor (DC/T cell ratio of 1:10) in serum-free X-VIVO-20. Proliferation of alloreactive T cells (5 × 104/well, triplicate cultures) was determined by [3H]TdR incorporation. A representative result of four independent experiments is shown.
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Figure 2: Proliferation of alloreactive CD4+ T cells induced by iDCs or mDCs. Naive CD4+ T cells purified by MACS sorting from cord blood were primed and restimulated (every week) with allogeneic iDCs or mDCs from the same donor (DC/T cell ratio of 1:10) in serum-free X-VIVO-20. Proliferation of alloreactive T cells (5 × 104/well, triplicate cultures) was determined by [3H]TdR incorporation. A representative result of four independent experiments is shown.

Mentions: mDCs as well as iDCs were used for repetitive stimulations of naive cord blood–derived CD4+ T cells. Priming and repetitive restimulations with mDCs resulted in a strong proliferation of alloreactive T cells (Fig. 2) and a 30–50-fold expansion after two rounds of restimulation. In contrast, iDCs induced only a weak proliferation of alloreactive T cells in the primary culture, and the antigen-driven proliferation further decreased in the successive restimulations (Fig. 2), finally resulting in nonproliferative CD4+ T cells. Therefore, it was not possible to induce stable alloreactive T cell lines or a significant expansion of these T cells with allogeneic iDCs. The number of T cells also decreased slowly after repetitive stimulations. 40–60% of the initial T cell numbers could be recovered after two rounds of restimulation. However, neither significant necrosis nor apoptosis was observed in the cultures by propidium iodide and annexin V staining (data not shown).


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)

Proliferation of alloreactive CD4+ T cells induced by iDCs or mDCs. Naive CD4+ T cells purified by MACS sorting from cord blood were primed and restimulated (every week) with allogeneic iDCs or mDCs from the same donor (DC/T cell ratio of 1:10) in serum-free X-VIVO-20. Proliferation of alloreactive T cells (5 × 104/well, triplicate cultures) was determined by [3H]TdR incorporation. A representative result of four independent experiments is shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193357&req=5

Figure 2: Proliferation of alloreactive CD4+ T cells induced by iDCs or mDCs. Naive CD4+ T cells purified by MACS sorting from cord blood were primed and restimulated (every week) with allogeneic iDCs or mDCs from the same donor (DC/T cell ratio of 1:10) in serum-free X-VIVO-20. Proliferation of alloreactive T cells (5 × 104/well, triplicate cultures) was determined by [3H]TdR incorporation. A representative result of four independent experiments is shown.
Mentions: mDCs as well as iDCs were used for repetitive stimulations of naive cord blood–derived CD4+ T cells. Priming and repetitive restimulations with mDCs resulted in a strong proliferation of alloreactive T cells (Fig. 2) and a 30–50-fold expansion after two rounds of restimulation. In contrast, iDCs induced only a weak proliferation of alloreactive T cells in the primary culture, and the antigen-driven proliferation further decreased in the successive restimulations (Fig. 2), finally resulting in nonproliferative CD4+ T cells. Therefore, it was not possible to induce stable alloreactive T cell lines or a significant expansion of these T cells with allogeneic iDCs. The number of T cells also decreased slowly after repetitive stimulations. 40–60% of the initial T cell numbers could be recovered after two rounds of restimulation. However, neither significant necrosis nor apoptosis was observed in the cultures by propidium iodide and annexin V staining (data not shown).

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