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Efficient targeting of protein antigen to the dendritic cell receptor DEC-205 in the steady state leads to antigen presentation on major histocompatibility complex class I products and peripheral CD8+ T cell tolerance.

Bonifaz L, Bonnyay D, Mahnke K, Rivera M, Nussenzweig MC, Steinman RM - J. Exp. Med. (2002)

Bottom Line: In vivo, the OVA protein was selectively presented by DCs to TCR transgenic CD8+ cells, again at least 400 times more effectively than soluble OVA and in a TAP-dependent fashion.In contrast, simultaneous delivery of a DC maturation stimulus via CD40, together with alphaDEC-205:OVA, induced strong immunity.Therefore, DEC-205 provides an efficient receptor-based mechanism for DCs to process proteins for MHC class I presentation in vivo, leading to tolerance in the steady state and immunity after DC maturation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, The Rockefeller University, New York, NY 10021, USA.

ABSTRACT
To identify endocytic receptors that allow dendritic cells (DCs) to capture and present antigens on major histocompatibility complex (MHC) class I products in vivo, we evaluated DEC-205, which is abundant on DCs in lymphoid tissues. Ovalbumin (OVA) protein, when chemically coupled to monoclonal alphaDEC-205 antibody, was presented by CD11c+ lymph node DCs, but not by CD11c- cells, to OVA-specific, CD4+ and CD8+ T cells. Receptor-mediated presentation was at least 400 times more efficient than unconjugated OVA and, for MHC class I, the DCs had to express transporter of antigenic peptides (TAP) transporters. When alphaDEC-205:OVA was injected subcutaneously, OVA protein was identified over a 4-48 h period in DCs, primarily in the lymph nodes draining the injection site. In vivo, the OVA protein was selectively presented by DCs to TCR transgenic CD8+ cells, again at least 400 times more effectively than soluble OVA and in a TAP-dependent fashion. Targeting of alphaDEC-205:OVA to DCs in the steady state initially induced 4-7 cycles of T cell division, but the T cells were then deleted and the mice became specifically unresponsive to rechallenge with OVA in complete Freund's adjuvant. In contrast, simultaneous delivery of a DC maturation stimulus via CD40, together with alphaDEC-205:OVA, induced strong immunity. The CD8+ T cells responding in the presence of agonistic alphaCD40 antibody produced large amounts of interleukin 2 and interferon gamma, acquired cytolytic function in vivo, emigrated in large numbers to the lung, and responded vigorously to OVA rechallenge. Therefore, DEC-205 provides an efficient receptor-based mechanism for DCs to process proteins for MHC class I presentation in vivo, leading to tolerance in the steady state and immunity after DC maturation.

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Contrasting responses of OT-I T cells to αDEC-205:OVA in the presence or absence of αCD40-induced DC maturation. (A) αCD40 has little impact on αDEC-205:OVA induced proliferation of OT-I T cells. As in Fig. 3 E, but mice were or were not given αCD40 (100 μg FGK45.5 subcutaneously). (B) Differential IL-2 and IFN-γ production by OT-I T cells in response to isotype:OVA or αDEC-205:OVA with or without αCD40. As in panel A, but lymph node suspensions were restimulated in vitro with the cognate OT-I peptide for 5 h in the presence of brefeldin A (5 μg/ml) before staining for intracellular cytokine. Data are representative of three experiments.
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fig5: Contrasting responses of OT-I T cells to αDEC-205:OVA in the presence or absence of αCD40-induced DC maturation. (A) αCD40 has little impact on αDEC-205:OVA induced proliferation of OT-I T cells. As in Fig. 3 E, but mice were or were not given αCD40 (100 μg FGK45.5 subcutaneously). (B) Differential IL-2 and IFN-γ production by OT-I T cells in response to isotype:OVA or αDEC-205:OVA with or without αCD40. As in panel A, but lymph node suspensions were restimulated in vitro with the cognate OT-I peptide for 5 h in the presence of brefeldin A (5 μg/ml) before staining for intracellular cytokine. Data are representative of three experiments.

Mentions: To follow the fate of the OT-I T cells that proliferated in response to antigen targeted to DCs in vivo, we tracked the transferred T cells by flow cytometry using a combination of CD45.1 and Vβ5.1/5.2 markers, and we compared responses in the steady state to those following αCD40-induced DC maturation. At 3 d after αDEC-205:OVA injection, we found strong proliferative responses in the presence or absence DC maturation (Fig. 5 A). However, αCD40 treated mice also showed greatly enhanced T cell production of IL-2 and IFN-γ (Fig. 5 B, bottom row).


Efficient targeting of protein antigen to the dendritic cell receptor DEC-205 in the steady state leads to antigen presentation on major histocompatibility complex class I products and peripheral CD8+ T cell tolerance.

Bonifaz L, Bonnyay D, Mahnke K, Rivera M, Nussenzweig MC, Steinman RM - J. Exp. Med. (2002)

Contrasting responses of OT-I T cells to αDEC-205:OVA in the presence or absence of αCD40-induced DC maturation. (A) αCD40 has little impact on αDEC-205:OVA induced proliferation of OT-I T cells. As in Fig. 3 E, but mice were or were not given αCD40 (100 μg FGK45.5 subcutaneously). (B) Differential IL-2 and IFN-γ production by OT-I T cells in response to isotype:OVA or αDEC-205:OVA with or without αCD40. As in panel A, but lymph node suspensions were restimulated in vitro with the cognate OT-I peptide for 5 h in the presence of brefeldin A (5 μg/ml) before staining for intracellular cytokine. Data are representative of three experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196060&req=5

fig5: Contrasting responses of OT-I T cells to αDEC-205:OVA in the presence or absence of αCD40-induced DC maturation. (A) αCD40 has little impact on αDEC-205:OVA induced proliferation of OT-I T cells. As in Fig. 3 E, but mice were or were not given αCD40 (100 μg FGK45.5 subcutaneously). (B) Differential IL-2 and IFN-γ production by OT-I T cells in response to isotype:OVA or αDEC-205:OVA with or without αCD40. As in panel A, but lymph node suspensions were restimulated in vitro with the cognate OT-I peptide for 5 h in the presence of brefeldin A (5 μg/ml) before staining for intracellular cytokine. Data are representative of three experiments.
Mentions: To follow the fate of the OT-I T cells that proliferated in response to antigen targeted to DCs in vivo, we tracked the transferred T cells by flow cytometry using a combination of CD45.1 and Vβ5.1/5.2 markers, and we compared responses in the steady state to those following αCD40-induced DC maturation. At 3 d after αDEC-205:OVA injection, we found strong proliferative responses in the presence or absence DC maturation (Fig. 5 A). However, αCD40 treated mice also showed greatly enhanced T cell production of IL-2 and IFN-γ (Fig. 5 B, bottom row).

Bottom Line: In vivo, the OVA protein was selectively presented by DCs to TCR transgenic CD8+ cells, again at least 400 times more effectively than soluble OVA and in a TAP-dependent fashion.In contrast, simultaneous delivery of a DC maturation stimulus via CD40, together with alphaDEC-205:OVA, induced strong immunity.Therefore, DEC-205 provides an efficient receptor-based mechanism for DCs to process proteins for MHC class I presentation in vivo, leading to tolerance in the steady state and immunity after DC maturation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, The Rockefeller University, New York, NY 10021, USA.

ABSTRACT
To identify endocytic receptors that allow dendritic cells (DCs) to capture and present antigens on major histocompatibility complex (MHC) class I products in vivo, we evaluated DEC-205, which is abundant on DCs in lymphoid tissues. Ovalbumin (OVA) protein, when chemically coupled to monoclonal alphaDEC-205 antibody, was presented by CD11c+ lymph node DCs, but not by CD11c- cells, to OVA-specific, CD4+ and CD8+ T cells. Receptor-mediated presentation was at least 400 times more efficient than unconjugated OVA and, for MHC class I, the DCs had to express transporter of antigenic peptides (TAP) transporters. When alphaDEC-205:OVA was injected subcutaneously, OVA protein was identified over a 4-48 h period in DCs, primarily in the lymph nodes draining the injection site. In vivo, the OVA protein was selectively presented by DCs to TCR transgenic CD8+ cells, again at least 400 times more effectively than soluble OVA and in a TAP-dependent fashion. Targeting of alphaDEC-205:OVA to DCs in the steady state initially induced 4-7 cycles of T cell division, but the T cells were then deleted and the mice became specifically unresponsive to rechallenge with OVA in complete Freund's adjuvant. In contrast, simultaneous delivery of a DC maturation stimulus via CD40, together with alphaDEC-205:OVA, induced strong immunity. The CD8+ T cells responding in the presence of agonistic alphaCD40 antibody produced large amounts of interleukin 2 and interferon gamma, acquired cytolytic function in vivo, emigrated in large numbers to the lung, and responded vigorously to OVA rechallenge. Therefore, DEC-205 provides an efficient receptor-based mechanism for DCs to process proteins for MHC class I presentation in vivo, leading to tolerance in the steady state and immunity after DC maturation.

Show MeSH
Related in: MedlinePlus