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In vitro generation of interleukin 10-producing regulatory CD4(+) T cells is induced by immunosuppressive drugs and inhibited by T helper type 1 (Th1)- and Th2-inducing cytokines.

Barrat FJ, Cua DJ, Boonstra A, Richards DF, Crain C, Savelkoul HF, de Waal-Malefyt R, Coffman RL, Hawrylowicz CM, O'Garra A - J. Exp. Med. (2002)

Bottom Line: Furthermore, nuclear factor (NF)-kappaB and activator protein (AP)-1 activities were inhibited in the IL-10-producing cells described here as well as key transcription factors involved in Th1 and Th2 subset differentiation.The regulatory function of these in vitro generated IL-10-producing T cells was demonstrated by their ability to prevent central nervous system inflammation, when targeted to the site of inflammation, and this function was shown to be IL-10 dependent.Generating homogeneous populations of IL-10-producing T cells in vitro will thus facilitate the use of regulatory T cells in immunotherapy.

View Article: PubMed Central - PubMed

Affiliation: DNAX Research Institute, Department of Immunology, Palo Alto, CA 94304, USA. fbarrat@dvax.com

ABSTRACT
We show that a combination of the immunosuppressive drugs, vitamin D3 and Dexamethasone, induced human and mouse naive CD4(+) T cells to differentiate in vitro into regulatory T cells. In contrast to the previously described in vitro derived CD4(+) T cells, these cells produced only interleukin (IL)-10, but no IL-5 and interferon (IFN)-gamma, and furthermore retained strong proliferative capacity. The development of these IL-10-producing cells was enhanced by neutralization of the T helper type 1 (Th1)- and Th2-inducing cytokines IL-4, IL-12, and IFN-gamma. These immunosuppressive drugs also induced the development of IL-10-producing T cells in the absence of antigen-presenting cells, with IL-10 acting as a positive autocrine factor for these T cells. Furthermore, nuclear factor (NF)-kappaB and activator protein (AP)-1 activities were inhibited in the IL-10-producing cells described here as well as key transcription factors involved in Th1 and Th2 subset differentiation. The regulatory function of these in vitro generated IL-10-producing T cells was demonstrated by their ability to prevent central nervous system inflammation, when targeted to the site of inflammation, and this function was shown to be IL-10 dependent. Generating homogeneous populations of IL-10-producing T cells in vitro will thus facilitate the use of regulatory T cells in immunotherapy.

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Related in: MedlinePlus

VitD3/Dex induces the development of human T cells producing IL-10 and no IL-4, IL-5, or IFN-γ. Purified human CD4+CD45RA+ were stimulated with plate-bound anti-CD3, soluble anti-CD28, and IL-2 in the presence of neutralizing anti–IL-4, anti–IFN-γ, and anti–IL-12 mAbs. After four rounds of stimulation, cells were characterized for cytokine production by immunoassay (A) as well as by intracellular flow cytometric analysis (B). Representative results of four experiments are shown.
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fig3: VitD3/Dex induces the development of human T cells producing IL-10 and no IL-4, IL-5, or IFN-γ. Purified human CD4+CD45RA+ were stimulated with plate-bound anti-CD3, soluble anti-CD28, and IL-2 in the presence of neutralizing anti–IL-4, anti–IFN-γ, and anti–IL-12 mAbs. After four rounds of stimulation, cells were characterized for cytokine production by immunoassay (A) as well as by intracellular flow cytometric analysis (B). Representative results of four experiments are shown.

Mentions: In view of the potential implications of our results regarding immunotherapy, conditions for generating IL-10–producing T cells in the mouse, using Vit/Dex and anti-CD3 and anti-CD28 stimulation in the complete absence of IL-4 and IFN-γ, were reproduced with human T cells. Naive human CD4+CD45RA+ T cells cultured under these conditions resulted in T cells producing high levels of IL-10 and no IL-4, IL-5, or IFN-γ (Fig. 3 A). This was also achieved in the presence of APC (data not shown). Similarly to our results using mouse T cells (Fig. 2 B), the combination of Vit/Dex led to an enhanced percentage of human IL-10–producing T cells, compared with the effects of Vit and Dex alone (Fig. 3 B). These cells did not produce any IL-4 or IFN-γ. The ability to generate human T cells producing IL-10 in the absence of Th1 or Th2-type cytokines, is a significant step toward their potential application in adoptive immunotherapy.


In vitro generation of interleukin 10-producing regulatory CD4(+) T cells is induced by immunosuppressive drugs and inhibited by T helper type 1 (Th1)- and Th2-inducing cytokines.

Barrat FJ, Cua DJ, Boonstra A, Richards DF, Crain C, Savelkoul HF, de Waal-Malefyt R, Coffman RL, Hawrylowicz CM, O'Garra A - J. Exp. Med. (2002)

VitD3/Dex induces the development of human T cells producing IL-10 and no IL-4, IL-5, or IFN-γ. Purified human CD4+CD45RA+ were stimulated with plate-bound anti-CD3, soluble anti-CD28, and IL-2 in the presence of neutralizing anti–IL-4, anti–IFN-γ, and anti–IL-12 mAbs. After four rounds of stimulation, cells were characterized for cytokine production by immunoassay (A) as well as by intracellular flow cytometric analysis (B). Representative results of four experiments are shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193760&req=5

fig3: VitD3/Dex induces the development of human T cells producing IL-10 and no IL-4, IL-5, or IFN-γ. Purified human CD4+CD45RA+ were stimulated with plate-bound anti-CD3, soluble anti-CD28, and IL-2 in the presence of neutralizing anti–IL-4, anti–IFN-γ, and anti–IL-12 mAbs. After four rounds of stimulation, cells were characterized for cytokine production by immunoassay (A) as well as by intracellular flow cytometric analysis (B). Representative results of four experiments are shown.
Mentions: In view of the potential implications of our results regarding immunotherapy, conditions for generating IL-10–producing T cells in the mouse, using Vit/Dex and anti-CD3 and anti-CD28 stimulation in the complete absence of IL-4 and IFN-γ, were reproduced with human T cells. Naive human CD4+CD45RA+ T cells cultured under these conditions resulted in T cells producing high levels of IL-10 and no IL-4, IL-5, or IFN-γ (Fig. 3 A). This was also achieved in the presence of APC (data not shown). Similarly to our results using mouse T cells (Fig. 2 B), the combination of Vit/Dex led to an enhanced percentage of human IL-10–producing T cells, compared with the effects of Vit and Dex alone (Fig. 3 B). These cells did not produce any IL-4 or IFN-γ. The ability to generate human T cells producing IL-10 in the absence of Th1 or Th2-type cytokines, is a significant step toward their potential application in adoptive immunotherapy.

Bottom Line: Furthermore, nuclear factor (NF)-kappaB and activator protein (AP)-1 activities were inhibited in the IL-10-producing cells described here as well as key transcription factors involved in Th1 and Th2 subset differentiation.The regulatory function of these in vitro generated IL-10-producing T cells was demonstrated by their ability to prevent central nervous system inflammation, when targeted to the site of inflammation, and this function was shown to be IL-10 dependent.Generating homogeneous populations of IL-10-producing T cells in vitro will thus facilitate the use of regulatory T cells in immunotherapy.

View Article: PubMed Central - PubMed

Affiliation: DNAX Research Institute, Department of Immunology, Palo Alto, CA 94304, USA. fbarrat@dvax.com

ABSTRACT
We show that a combination of the immunosuppressive drugs, vitamin D3 and Dexamethasone, induced human and mouse naive CD4(+) T cells to differentiate in vitro into regulatory T cells. In contrast to the previously described in vitro derived CD4(+) T cells, these cells produced only interleukin (IL)-10, but no IL-5 and interferon (IFN)-gamma, and furthermore retained strong proliferative capacity. The development of these IL-10-producing cells was enhanced by neutralization of the T helper type 1 (Th1)- and Th2-inducing cytokines IL-4, IL-12, and IFN-gamma. These immunosuppressive drugs also induced the development of IL-10-producing T cells in the absence of antigen-presenting cells, with IL-10 acting as a positive autocrine factor for these T cells. Furthermore, nuclear factor (NF)-kappaB and activator protein (AP)-1 activities were inhibited in the IL-10-producing cells described here as well as key transcription factors involved in Th1 and Th2 subset differentiation. The regulatory function of these in vitro generated IL-10-producing T cells was demonstrated by their ability to prevent central nervous system inflammation, when targeted to the site of inflammation, and this function was shown to be IL-10 dependent. Generating homogeneous populations of IL-10-producing T cells in vitro will thus facilitate the use of regulatory T cells in immunotherapy.

Show MeSH
Related in: MedlinePlus