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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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γ/δ clone JR.2 promotes DC antigen presentation to CD4+ T cells. (A) Allogeneic MLR. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone. Irradiated DC were then cultured at DC:T cell ratios from 1:1,000 to 1:10 with allogeneic CD4+ T cells for 7 d and proliferation was measured. DCs that had been matured in the presence of TNF-α or γ/δ clone JR.2 resulted in increased proliferation of allogeneic CD4+ T cells when compared with immature DCs. Squares, medium alone; inverted triangles, TNF-α; diamonds, JR.2. (B) KLH antigen presentation. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone in the presence or absence of 25 μg/ml Keyhole Limpet Hemacyanin (KLH). Irradiated DCs were then cultured with 5 × 104 autologous CD4+CD45RA+ naive T cells/well at a DC:T cell ratio of 1:10 for 5 d and proliferation was measured. Only DCs pulsed with KLH and matured in the presence of TNFα or γ/δ clone JR.2 resulted in proliferation by naive responder T cells. Results are expressed as relative stimulation index. Results are representative of two independent experiments using different DC donors.
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fig7: γ/δ clone JR.2 promotes DC antigen presentation to CD4+ T cells. (A) Allogeneic MLR. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone. Irradiated DC were then cultured at DC:T cell ratios from 1:1,000 to 1:10 with allogeneic CD4+ T cells for 7 d and proliferation was measured. DCs that had been matured in the presence of TNF-α or γ/δ clone JR.2 resulted in increased proliferation of allogeneic CD4+ T cells when compared with immature DCs. Squares, medium alone; inverted triangles, TNF-α; diamonds, JR.2. (B) KLH antigen presentation. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone in the presence or absence of 25 μg/ml Keyhole Limpet Hemacyanin (KLH). Irradiated DCs were then cultured with 5 × 104 autologous CD4+CD45RA+ naive T cells/well at a DC:T cell ratio of 1:10 for 5 d and proliferation was measured. Only DCs pulsed with KLH and matured in the presence of TNFα or γ/δ clone JR.2 resulted in proliferation by naive responder T cells. Results are expressed as relative stimulation index. Results are representative of two independent experiments using different DC donors.

Mentions: One hallmark of mature DCs is their ability to present antigen to naive T cells encountered in specialized locations such as the draining lymph node. This interaction is critical for the development of an adaptive immune response and the generation of immunologic memory. We employed a system to determine if DCs, matured in the presence of the γ/δ T cell clone JR.2, were able to efficiently present antigens to CD4+ T cells. As mature DCs display an increased ability to stimulate allogeneic CD4+ T cells in a mixed lymphocyte reaction (MLR), we first assessed the capacity of DCs matured in the presence of γ/δ T cell clone JR.2 (T cell:DC ratio = 1:3) or TNF-α (50 ng/ml) to stimulate allogeneic CD4+ T cells. CD4+ T cells were cultured for 7 d with irradiated allogeneic immature monocyte-derived DCs that had been previously cultured for 72 h in the presence of TNF-α, JR.2, or medium alone. During the final 12 h of culture, cells were pulsed with [3H]thymidine and assayed for proliferation. DCs matured in the presence of JR.2 or TNF-α led to increased proliferation by allogeneic CD4+ T cells when compared with immature DCs that had been cultured in medium alone (Fig. 7 A). We next employed a system to determine if DCs, matured in the presence of the γ/δ T cell clone JR.2, were able to efficiently present KLH, an antigen encountered for the first time, to naive T cells. Human CD4+CD45RA+ naive T cells were isolated using a negative selection mAb column. Autologous immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α (50 ng/ml), γ/δ clone JR.2 (T cell:DC ratio = 1:10), or medium alone with or without 25 μg/ml KLH. DCs cultured in either TNF-α (54% CD83 positive, CD86 MFI = 608) or the presence of JR.2 (42% CD83 positive, CD86 MFI = 664) expressed increased levels of CD83 and CD86 consistent with a mature DC phenotype. In contrast, the presence of KLH had no effect upon phenotypic maturation (2.5% CD83 positive, CD86 MFI = 16; data not depicted). On day 3 of culture, these DCs were washed, irradiated (5,000 rads), and then cultured with autologous naive T cells at a DC:T cell ratio of 1 to 10 for 7 d. During the final 12 h of culture, cells were pulsed with [3H]thymidine and assayed for proliferation. Only DCs pulsed with KLH and matured in the presence of TNF-α (SI = 3.6) or the CD1c-restricted γ/δ T cell clone JR.2 (SI = 5) led to specific proliferation by the naive T cells reactive against KLH (Fig. 7 B).


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)

γ/δ clone JR.2 promotes DC antigen presentation to CD4+ T cells. (A) Allogeneic MLR. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone. Irradiated DC were then cultured at DC:T cell ratios from 1:1,000 to 1:10 with allogeneic CD4+ T cells for 7 d and proliferation was measured. DCs that had been matured in the presence of TNF-α or γ/δ clone JR.2 resulted in increased proliferation of allogeneic CD4+ T cells when compared with immature DCs. Squares, medium alone; inverted triangles, TNF-α; diamonds, JR.2. (B) KLH antigen presentation. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone in the presence or absence of 25 μg/ml Keyhole Limpet Hemacyanin (KLH). Irradiated DCs were then cultured with 5 × 104 autologous CD4+CD45RA+ naive T cells/well at a DC:T cell ratio of 1:10 for 5 d and proliferation was measured. Only DCs pulsed with KLH and matured in the presence of TNFα or γ/δ clone JR.2 resulted in proliferation by naive responder T cells. Results are expressed as relative stimulation index. Results are representative of two independent experiments using different DC donors.
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Related In: Results  -  Collection

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fig7: γ/δ clone JR.2 promotes DC antigen presentation to CD4+ T cells. (A) Allogeneic MLR. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone. Irradiated DC were then cultured at DC:T cell ratios from 1:1,000 to 1:10 with allogeneic CD4+ T cells for 7 d and proliferation was measured. DCs that had been matured in the presence of TNF-α or γ/δ clone JR.2 resulted in increased proliferation of allogeneic CD4+ T cells when compared with immature DCs. Squares, medium alone; inverted triangles, TNF-α; diamonds, JR.2. (B) KLH antigen presentation. Immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α, γ/δ clone JR.2, or medium alone in the presence or absence of 25 μg/ml Keyhole Limpet Hemacyanin (KLH). Irradiated DCs were then cultured with 5 × 104 autologous CD4+CD45RA+ naive T cells/well at a DC:T cell ratio of 1:10 for 5 d and proliferation was measured. Only DCs pulsed with KLH and matured in the presence of TNFα or γ/δ clone JR.2 resulted in proliferation by naive responder T cells. Results are expressed as relative stimulation index. Results are representative of two independent experiments using different DC donors.
Mentions: One hallmark of mature DCs is their ability to present antigen to naive T cells encountered in specialized locations such as the draining lymph node. This interaction is critical for the development of an adaptive immune response and the generation of immunologic memory. We employed a system to determine if DCs, matured in the presence of the γ/δ T cell clone JR.2, were able to efficiently present antigens to CD4+ T cells. As mature DCs display an increased ability to stimulate allogeneic CD4+ T cells in a mixed lymphocyte reaction (MLR), we first assessed the capacity of DCs matured in the presence of γ/δ T cell clone JR.2 (T cell:DC ratio = 1:3) or TNF-α (50 ng/ml) to stimulate allogeneic CD4+ T cells. CD4+ T cells were cultured for 7 d with irradiated allogeneic immature monocyte-derived DCs that had been previously cultured for 72 h in the presence of TNF-α, JR.2, or medium alone. During the final 12 h of culture, cells were pulsed with [3H]thymidine and assayed for proliferation. DCs matured in the presence of JR.2 or TNF-α led to increased proliferation by allogeneic CD4+ T cells when compared with immature DCs that had been cultured in medium alone (Fig. 7 A). We next employed a system to determine if DCs, matured in the presence of the γ/δ T cell clone JR.2, were able to efficiently present KLH, an antigen encountered for the first time, to naive T cells. Human CD4+CD45RA+ naive T cells were isolated using a negative selection mAb column. Autologous immature monocyte-derived DCs were cultured for 72 h in the presence of TNF-α (50 ng/ml), γ/δ clone JR.2 (T cell:DC ratio = 1:10), or medium alone with or without 25 μg/ml KLH. DCs cultured in either TNF-α (54% CD83 positive, CD86 MFI = 608) or the presence of JR.2 (42% CD83 positive, CD86 MFI = 664) expressed increased levels of CD83 and CD86 consistent with a mature DC phenotype. In contrast, the presence of KLH had no effect upon phenotypic maturation (2.5% CD83 positive, CD86 MFI = 16; data not depicted). On day 3 of culture, these DCs were washed, irradiated (5,000 rads), and then cultured with autologous naive T cells at a DC:T cell ratio of 1 to 10 for 7 d. During the final 12 h of culture, cells were pulsed with [3H]thymidine and assayed for proliferation. Only DCs pulsed with KLH and matured in the presence of TNF-α (SI = 3.6) or the CD1c-restricted γ/δ T cell clone JR.2 (SI = 5) led to specific proliferation by the naive T cells reactive against KLH (Fig. 7 B).

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