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CD1-mediated gamma/delta T cell maturation of dendritic cells.

Leslie DS, Vincent MS, Spada FM, Das H, Sugita M, Morita CT, Brenner MB - J. Exp. Med. (2002)

Bottom Line: In addition, these DCs were able to efficiently present peptide antigens to naive CD4+ T cells.CD1-restricted gamma/delta T cell recognition of immature DCs provides the human immune system with the capacity to rapidly generate a pool of mature DCs early during microbial invasion.This may be an important source of critical host signals for T helper type 1 polarization of antigen-specific naive T cells and the subsequent adaptive immune response.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women's Hospital at Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Immature myeloid dendritic cells (DCs) express only low levels of major histocompatibility complex (MHC) class II but express high levels of CD1 a, b, and c antigen-presenting molecules at the cell surface. As Vdelta1+ gamma/delta T cells are the main tissue subset of gamma/delta T cells and they are known to recognize CD1c in the absence of specific foreign antigen recognition, we examined the possible interaction of these T cells with immature DCs. We show that CD1-restricted gamma/delta T cells can mediate the maturation of DCs. DC maturation required cell-cell contact and could be blocked by antibodies against CD1c. The maturation process was partially mediated by tumor necrosis factor alpha. Importantly, immature DCs matured in the presence of lipopolysaccharide and CD1-restricted gamma/delta T cells produced bioactive interleukin-12p70. In addition, these DCs were able to efficiently present peptide antigens to naive CD4+ T cells. CD1-restricted gamma/delta T cell recognition of immature DCs provides the human immune system with the capacity to rapidly generate a pool of mature DCs early during microbial invasion. This may be an important source of critical host signals for T helper type 1 polarization of antigen-specific naive T cells and the subsequent adaptive immune response.

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Role of γ/δ T cell surface molecules and soluble mediators in DC maturation. (A) CD40 ligand cell surface expression by JR.2. γ/δ T cells. CD40 ligand expression on the surface of γ/δ T cell clone JR.2 increased approximately fourfold during a 24 h coculture with immature DCs as assessed by mAb staining and flow cytometry. Triangles, culture with DC; squares, medium control. (B) Inhibition of DC maturation by blocking of γ/δ T cell surface molecules and the cytokines TNF-α and IFN-γ. Immature DCs were cocultured with LPS (10 ng/ml) or JR.2 for 48 h. In the JR.2:DC cocultures the presence of mAb against TNF-α inhibited up-regulation of DCs expressing CD83 by ∼50%, while culture in the presence of mAbs against IFN-γ, CD40L, FasL, or the RANKL antagonist OPG had no effect. These results are representative of four independent experiments using different DC donors.
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fig5: Role of γ/δ T cell surface molecules and soluble mediators in DC maturation. (A) CD40 ligand cell surface expression by JR.2. γ/δ T cells. CD40 ligand expression on the surface of γ/δ T cell clone JR.2 increased approximately fourfold during a 24 h coculture with immature DCs as assessed by mAb staining and flow cytometry. Triangles, culture with DC; squares, medium control. (B) Inhibition of DC maturation by blocking of γ/δ T cell surface molecules and the cytokines TNF-α and IFN-γ. Immature DCs were cocultured with LPS (10 ng/ml) or JR.2 for 48 h. In the JR.2:DC cocultures the presence of mAb against TNF-α inhibited up-regulation of DCs expressing CD83 by ∼50%, while culture in the presence of mAbs against IFN-γ, CD40L, FasL, or the RANKL antagonist OPG had no effect. These results are representative of four independent experiments using different DC donors.

Mentions: The Th1 cytokines IFN-γ (19) and TNF-α (20) have been extensively characterized as soluble factors that are important in DC maturation. For example, production of IFN-γ by host lymphocytes including CD8+ T cells (21) and NK cells (22) was found to promote the ability of maturing DCs to produce IL-12p70 leading to Th1 polarization of antigen specific naive T cells. Therefore, we assessed the ability of γ/δ clone JR.2 to produce these cytokines during coculture with CD1 expressing immature DCs. Both IFN-γ (1.4 ± 0.05 ng/ml) and TNF-α (14.3 ± 3.3 ng/ml) were produced by CD1-specific γ/δ T cells as assessed by ELISA, and cytokine production was inhibited by 89% using mAb against CD1c but not by mAb against CD1a (Fig. 4 A). Production of the Th2 cytokines IL-4 and IL-13 was below the limit of detection (unpublished data). We next examined the role of these cytokines in DC maturation. The addition of anti–TNF-α mAb at 20 μg/ml partially inhibited phenotypic maturation of immature DCs cultured with JR.2 (70% decrease in CD83-positive cells compared with no antibody blocking) while addition of anti–IFN-γ at 20 μg/ml did not inhibit DC maturation as assessed by cell surface expression of CD83 and CD86 using flow cytometry (Fig. 4 B). These results are consistent with production of a soluble factor such as TNF-α being partially responsible for the CD1-mediated maturation of DC by γ/δ T cells. We next employed a transwell system to determine if cell–cell contact was required for maturation to occur. Immature DCs were cultured in direct contact with the JR.2 γ/δ clone or were separated by a 0.2 μ transwell membrane. DCs separated from JR.2 T cells by the transwell failed to up-regulate CD83. To assess the specific role of soluble factors, DCs and JR.2 were cocultured together (T cell:DC ratio = 1:10) and additional DCs separated in the transwell were assayed for CD83 up-regulation. The percentage of CD83+ cells among the DCs exposed to soluble factors in this transwell increased twofold (20 to 40% positive), whereas DCs cultured in direct contact with γ/δ clone JR.2 in the absence of a transwell demonstrated a fourfold increase (20 to 80% positive; data not depicted). These results are consistent with production of a soluble factor such as TNFα being only partially responsible for the CD1-mediated maturation of DCs by γ/δ T cells. Other investigators have emphasized the role of cell surface TNF family members such as CD40 ligand (CD154; reference 6), RANK ligand (7), and Fas ligand (8) expressed by T cells in the maturation of DCs. As rapid, tightly regulated expression of these cell surface T cell molecules may provide critical signals for DC maturation, we examined the expression of CD40 ligand by γ/δ clone JR.2 during culture with immature DCs. CD40 ligand cell surface levels on JR.2 T cells increased approximately fourfold (from MFI = 21.3 to 84.23) when cultured with CD1c expressing immature DCs over 24 h when compared with JR.2 T cells cultured in tissue culture medium alone, implying a potential role for CD40 ligand in the maturation process (Fig. 5 A). Although CD40 ligand expression on the surface of the JR.2 T cells increased during coculture with immature DCs, mAb blocking studies using antibodies against CD40L led to only 20% inhibition of CD83 up-regulation. In addition, mAb blocking of FasL, as well as the RANKL antagonist OPG, also failed to significantly inhibit maturation of DC as assessed by CD83 cell surface expression (Fig. 5 B). Taken together, our results emphasize the importance of soluble TNF-α production for the maturation of DCs by the CD1-reactive γ/δ clone JR.2. In addition, cell–cell contact between the DCs and CD1-restricted γ/δ T cell clone appears to be required for optimal maturation to occur, although we could not inhibit maturation with blockage of FasL or RANKL and we observed only partial inhibition via CD40L.


CD1-mediated gamma/delta T cell maturation of dendritic cells.

Leslie DS, Vincent MS, Spada FM, Das H, Sugita M, Morita CT, Brenner MB - J. Exp. Med. (2002)

Role of γ/δ T cell surface molecules and soluble mediators in DC maturation. (A) CD40 ligand cell surface expression by JR.2. γ/δ T cells. CD40 ligand expression on the surface of γ/δ T cell clone JR.2 increased approximately fourfold during a 24 h coculture with immature DCs as assessed by mAb staining and flow cytometry. Triangles, culture with DC; squares, medium control. (B) Inhibition of DC maturation by blocking of γ/δ T cell surface molecules and the cytokines TNF-α and IFN-γ. Immature DCs were cocultured with LPS (10 ng/ml) or JR.2 for 48 h. In the JR.2:DC cocultures the presence of mAb against TNF-α inhibited up-regulation of DCs expressing CD83 by ∼50%, while culture in the presence of mAbs against IFN-γ, CD40L, FasL, or the RANKL antagonist OPG had no effect. These results are representative of four independent experiments using different DC donors.
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Related In: Results  -  Collection

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fig5: Role of γ/δ T cell surface molecules and soluble mediators in DC maturation. (A) CD40 ligand cell surface expression by JR.2. γ/δ T cells. CD40 ligand expression on the surface of γ/δ T cell clone JR.2 increased approximately fourfold during a 24 h coculture with immature DCs as assessed by mAb staining and flow cytometry. Triangles, culture with DC; squares, medium control. (B) Inhibition of DC maturation by blocking of γ/δ T cell surface molecules and the cytokines TNF-α and IFN-γ. Immature DCs were cocultured with LPS (10 ng/ml) or JR.2 for 48 h. In the JR.2:DC cocultures the presence of mAb against TNF-α inhibited up-regulation of DCs expressing CD83 by ∼50%, while culture in the presence of mAbs against IFN-γ, CD40L, FasL, or the RANKL antagonist OPG had no effect. These results are representative of four independent experiments using different DC donors.
Mentions: The Th1 cytokines IFN-γ (19) and TNF-α (20) have been extensively characterized as soluble factors that are important in DC maturation. For example, production of IFN-γ by host lymphocytes including CD8+ T cells (21) and NK cells (22) was found to promote the ability of maturing DCs to produce IL-12p70 leading to Th1 polarization of antigen specific naive T cells. Therefore, we assessed the ability of γ/δ clone JR.2 to produce these cytokines during coculture with CD1 expressing immature DCs. Both IFN-γ (1.4 ± 0.05 ng/ml) and TNF-α (14.3 ± 3.3 ng/ml) were produced by CD1-specific γ/δ T cells as assessed by ELISA, and cytokine production was inhibited by 89% using mAb against CD1c but not by mAb against CD1a (Fig. 4 A). Production of the Th2 cytokines IL-4 and IL-13 was below the limit of detection (unpublished data). We next examined the role of these cytokines in DC maturation. The addition of anti–TNF-α mAb at 20 μg/ml partially inhibited phenotypic maturation of immature DCs cultured with JR.2 (70% decrease in CD83-positive cells compared with no antibody blocking) while addition of anti–IFN-γ at 20 μg/ml did not inhibit DC maturation as assessed by cell surface expression of CD83 and CD86 using flow cytometry (Fig. 4 B). These results are consistent with production of a soluble factor such as TNF-α being partially responsible for the CD1-mediated maturation of DC by γ/δ T cells. We next employed a transwell system to determine if cell–cell contact was required for maturation to occur. Immature DCs were cultured in direct contact with the JR.2 γ/δ clone or were separated by a 0.2 μ transwell membrane. DCs separated from JR.2 T cells by the transwell failed to up-regulate CD83. To assess the specific role of soluble factors, DCs and JR.2 were cocultured together (T cell:DC ratio = 1:10) and additional DCs separated in the transwell were assayed for CD83 up-regulation. The percentage of CD83+ cells among the DCs exposed to soluble factors in this transwell increased twofold (20 to 40% positive), whereas DCs cultured in direct contact with γ/δ clone JR.2 in the absence of a transwell demonstrated a fourfold increase (20 to 80% positive; data not depicted). These results are consistent with production of a soluble factor such as TNFα being only partially responsible for the CD1-mediated maturation of DCs by γ/δ T cells. Other investigators have emphasized the role of cell surface TNF family members such as CD40 ligand (CD154; reference 6), RANK ligand (7), and Fas ligand (8) expressed by T cells in the maturation of DCs. As rapid, tightly regulated expression of these cell surface T cell molecules may provide critical signals for DC maturation, we examined the expression of CD40 ligand by γ/δ clone JR.2 during culture with immature DCs. CD40 ligand cell surface levels on JR.2 T cells increased approximately fourfold (from MFI = 21.3 to 84.23) when cultured with CD1c expressing immature DCs over 24 h when compared with JR.2 T cells cultured in tissue culture medium alone, implying a potential role for CD40 ligand in the maturation process (Fig. 5 A). Although CD40 ligand expression on the surface of the JR.2 T cells increased during coculture with immature DCs, mAb blocking studies using antibodies against CD40L led to only 20% inhibition of CD83 up-regulation. In addition, mAb blocking of FasL, as well as the RANKL antagonist OPG, also failed to significantly inhibit maturation of DC as assessed by CD83 cell surface expression (Fig. 5 B). Taken together, our results emphasize the importance of soluble TNF-α production for the maturation of DCs by the CD1-reactive γ/δ clone JR.2. In addition, cell–cell contact between the DCs and CD1-restricted γ/δ T cell clone appears to be required for optimal maturation to occur, although we could not inhibit maturation with blockage of FasL or RANKL and we observed only partial inhibition via CD40L.

Bottom Line: In addition, these DCs were able to efficiently present peptide antigens to naive CD4+ T cells.CD1-restricted gamma/delta T cell recognition of immature DCs provides the human immune system with the capacity to rapidly generate a pool of mature DCs early during microbial invasion.This may be an important source of critical host signals for T helper type 1 polarization of antigen-specific naive T cells and the subsequent adaptive immune response.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women's Hospital at Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT
Immature myeloid dendritic cells (DCs) express only low levels of major histocompatibility complex (MHC) class II but express high levels of CD1 a, b, and c antigen-presenting molecules at the cell surface. As Vdelta1+ gamma/delta T cells are the main tissue subset of gamma/delta T cells and they are known to recognize CD1c in the absence of specific foreign antigen recognition, we examined the possible interaction of these T cells with immature DCs. We show that CD1-restricted gamma/delta T cells can mediate the maturation of DCs. DC maturation required cell-cell contact and could be blocked by antibodies against CD1c. The maturation process was partially mediated by tumor necrosis factor alpha. Importantly, immature DCs matured in the presence of lipopolysaccharide and CD1-restricted gamma/delta T cells produced bioactive interleukin-12p70. In addition, these DCs were able to efficiently present peptide antigens to naive CD4+ T cells. CD1-restricted gamma/delta T cell recognition of immature DCs provides the human immune system with the capacity to rapidly generate a pool of mature DCs early during microbial invasion. This may be an important source of critical host signals for T helper type 1 polarization of antigen-specific naive T cells and the subsequent adaptive immune response.

Show MeSH
Related in: MedlinePlus