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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.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.

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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Deletion of OT-I T cells in response to αDEC-205:OVA in steady state. (A) C57BL/6 mice were given CD45.1+ OT-I T cells and antigen as described in 3E with or without αCD40. 3 and 12 d later, lymph nodes, spleen, and blood were harvested and evaluated for OT-I T cells (CD45.1+Vβ5.1/5.2+) by flow cytometry. (B) Data on the number of OT-I cells, expressed as the mean percentage of CD8+ T cells from three experiments of the type shown in panel A. (C) αCD40-rescued OT-I T cells are primed and secrete IFN-γ. C57BL/6 mice were given OT-I T cells and antigen as described in panel A. 12 d after antigen administration, lymph nodes were harvested and OT-I T cells evaluated for IFN-γ secretion as described in Fig. 5 B. (D) OT-I T cells are not present in a peripheral non-lymphoid tissue, after presentation of αDEC-205:OVA by DCs in the steady state, in the absence of αCD40 stimulation. As in panel A, but the lung was harvested 10 d after antigen administration and the OT-I cells were evaluated for expression of CD62L and CD45.1.
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fig6: Deletion of OT-I T cells in response to αDEC-205:OVA in steady state. (A) C57BL/6 mice were given CD45.1+ OT-I T cells and antigen as described in 3E with or without αCD40. 3 and 12 d later, lymph nodes, spleen, and blood were harvested and evaluated for OT-I T cells (CD45.1+Vβ5.1/5.2+) by flow cytometry. (B) Data on the number of OT-I cells, expressed as the mean percentage of CD8+ T cells from three experiments of the type shown in panel A. (C) αCD40-rescued OT-I T cells are primed and secrete IFN-γ. C57BL/6 mice were given OT-I T cells and antigen as described in panel A. 12 d after antigen administration, lymph nodes were harvested and OT-I T cells evaluated for IFN-γ secretion as described in Fig. 5 B. (D) OT-I T cells are not present in a peripheral non-lymphoid tissue, after presentation of αDEC-205:OVA by DCs in the steady state, in the absence of αCD40 stimulation. As in panel A, but the lung was harvested 10 d after antigen administration and the OT-I cells were evaluated for expression of CD62L and CD45.1.

Mentions: When we followed the numbers of injected OT-I T cells at 3 d and at 12–14 d in several lymphoid tissues (Fig. 6, A and B) , we found that αDEC-205:OVA in the steady state first expanded the OT-I cells, but by 12–14 d, the T cells were virtually entirely absent from lymph nodes, spleen, or blood (Fig. 6 A, compare left and right panels, and Fig. 6 B). However, if the αDEC-205:OVA conjugate was given with αCD40, the OT-I cells expanded relative to the PBS control (Fig. 6, A and B) or isotype-control:OVA conjugate (data not depicted). Similarly, when IFN-γ production was monitored by FACS® (Fig. 6 C) or by ELISPOT assays (data not depicted), the combination of αDEC-205:OVA and αCD40 induced a strong expansion of cytokine producing effectors, whereas cytokine producing OT-I cells were virtually deleted from the lymph node when mice had received αDEC-205:OVA (Fig. 6 C). As it was possible that T cells were being redistributed to tissues in the steady state rather than undergoing true deletion, we isolated cells from one large peripheral organ, the lung. In mice exposed to αDEC-205:OVA, we could not find any OT-I T cells in the lung at day 10, but many OT-I cells were found in the lungs of mice given αDEC-205:OVA plus αCD40 (Fig. 6 D). The expanded numbers of cells in the lymph nodes and lungs in response to αDEC-205:OVA plus αCD40 included a large fraction of cells lacking the CD62L selectin for lymph node homing, a typical feature of activated T cells (Fig. 6 D). In summary, delivery of protein antigens to DCs leads to the deletion of MHC class I–restricted T cells, but antigen delivered together with a maturation stimulus, leads to T cell expansion, production of IFN-γ, and export of T cells to peripheral tissues.


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)

Deletion of OT-I T cells in response to αDEC-205:OVA in steady state. (A) C57BL/6 mice were given CD45.1+ OT-I T cells and antigen as described in 3E with or without αCD40. 3 and 12 d later, lymph nodes, spleen, and blood were harvested and evaluated for OT-I T cells (CD45.1+Vβ5.1/5.2+) by flow cytometry. (B) Data on the number of OT-I cells, expressed as the mean percentage of CD8+ T cells from three experiments of the type shown in panel A. (C) αCD40-rescued OT-I T cells are primed and secrete IFN-γ. C57BL/6 mice were given OT-I T cells and antigen as described in panel A. 12 d after antigen administration, lymph nodes were harvested and OT-I T cells evaluated for IFN-γ secretion as described in Fig. 5 B. (D) OT-I T cells are not present in a peripheral non-lymphoid tissue, after presentation of αDEC-205:OVA by DCs in the steady state, in the absence of αCD40 stimulation. As in panel A, but the lung was harvested 10 d after antigen administration and the OT-I cells were evaluated for expression of CD62L and CD45.1.
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fig6: Deletion of OT-I T cells in response to αDEC-205:OVA in steady state. (A) C57BL/6 mice were given CD45.1+ OT-I T cells and antigen as described in 3E with or without αCD40. 3 and 12 d later, lymph nodes, spleen, and blood were harvested and evaluated for OT-I T cells (CD45.1+Vβ5.1/5.2+) by flow cytometry. (B) Data on the number of OT-I cells, expressed as the mean percentage of CD8+ T cells from three experiments of the type shown in panel A. (C) αCD40-rescued OT-I T cells are primed and secrete IFN-γ. C57BL/6 mice were given OT-I T cells and antigen as described in panel A. 12 d after antigen administration, lymph nodes were harvested and OT-I T cells evaluated for IFN-γ secretion as described in Fig. 5 B. (D) OT-I T cells are not present in a peripheral non-lymphoid tissue, after presentation of αDEC-205:OVA by DCs in the steady state, in the absence of αCD40 stimulation. As in panel A, but the lung was harvested 10 d after antigen administration and the OT-I cells were evaluated for expression of CD62L and CD45.1.
Mentions: When we followed the numbers of injected OT-I T cells at 3 d and at 12–14 d in several lymphoid tissues (Fig. 6, A and B) , we found that αDEC-205:OVA in the steady state first expanded the OT-I cells, but by 12–14 d, the T cells were virtually entirely absent from lymph nodes, spleen, or blood (Fig. 6 A, compare left and right panels, and Fig. 6 B). However, if the αDEC-205:OVA conjugate was given with αCD40, the OT-I cells expanded relative to the PBS control (Fig. 6, A and B) or isotype-control:OVA conjugate (data not depicted). Similarly, when IFN-γ production was monitored by FACS® (Fig. 6 C) or by ELISPOT assays (data not depicted), the combination of αDEC-205:OVA and αCD40 induced a strong expansion of cytokine producing effectors, whereas cytokine producing OT-I cells were virtually deleted from the lymph node when mice had received αDEC-205:OVA (Fig. 6 C). As it was possible that T cells were being redistributed to tissues in the steady state rather than undergoing true deletion, we isolated cells from one large peripheral organ, the lung. In mice exposed to αDEC-205:OVA, we could not find any OT-I T cells in the lung at day 10, but many OT-I cells were found in the lungs of mice given αDEC-205:OVA plus αCD40 (Fig. 6 D). The expanded numbers of cells in the lymph nodes and lungs in response to αDEC-205:OVA plus αCD40 included a large fraction of cells lacking the CD62L selectin for lymph node homing, a typical feature of activated T cells (Fig. 6 D). In summary, delivery of protein antigens to DCs leads to the deletion of MHC class I–restricted T cells, but antigen delivered together with a maturation stimulus, leads to T cell expansion, production of IFN-γ, and export of T cells to peripheral tissues.

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.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.

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