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Constitutive versus activation-dependent cross-presentation of immune complexes by CD8(+) and CD8(-) dendritic cells in vivo.

den Haan JM, Bevan MJ - J. Exp. Med. (2002)

Bottom Line: Murine splenic dendritic cells (DCs) can be divided into two subsets based on CD8alpha expression, but the specific role of each subset in stimulation of T cells is largely unknown.An important function of DCs is the ability to take up exogenous antigens and cross-present them in the context of major histocompatibility complex (MHC) class I molecules to CD8(+) T cells.These results suggest that although CD8(+) DCs constitutively cross-present exogenous antigens in the context of MHC class I molecules, CD8(-) DCs only do so after activation, such as via ligation of Fc(gamma)Rs.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195-7370, USA.

ABSTRACT
Murine splenic dendritic cells (DCs) can be divided into two subsets based on CD8alpha expression, but the specific role of each subset in stimulation of T cells is largely unknown. An important function of DCs is the ability to take up exogenous antigens and cross-present them in the context of major histocompatibility complex (MHC) class I molecules to CD8(+) T cells. We previously demonstrated that, when cell-associated ovalbumin (OVA) is injected into mice, only the CD8(+) DC subset cross-presents OVA in the context of MHC class I. In contrast to this selectivity with cell-associated antigen, we show here that both DC subsets isolated from mice injected with OVA/anti-OVA immune complexes (OVA-IC) cross-present OVA to CD8(+) T cells. The use of immunoglobulin G Fc receptor (Fc(gamma)R) common gamma-chain-deficient mice revealed that the cross-presentation by CD8(-) DCs depended on the expression of gamma-chain-containing activating FcgammaRs, whereas cross-presentation by CD8(+) DCs was not reduced in gamma-chain-deficient mice. These results suggest that although CD8(+) DCs constitutively cross-present exogenous antigens in the context of MHC class I molecules, CD8(-) DCs only do so after activation, such as via ligation of Fc(gamma)Rs. Cross-presentation of immune complexes may play an important role in autoimmune diseases and the therapeutic effect of antitumor antibodies.

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DCs present OVA epitopes in association with both MHC class I and II after in vivo injection of OVA-IC. B6 mice were primed with 50 μg soluble OVA, 50 μg OVA incubated with 200 μg anti-HRP antibodies, or 50 μg OVA with 200 μg anti-OVA antibodies (OVA-IC). CD11c+ DCs were isolated 14 h after injection and analyzed for their ability to stimulate T cells in vitro. The indicated numbers of DCs were coincubated with RAG1-deficient OT-I cells (A) or purified CD4+ OT-II cells (B). Proliferation of cells was determined by [3H]thymidine incorporation. Error bars indicate SEM of triplicate wells.
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fig1: DCs present OVA epitopes in association with both MHC class I and II after in vivo injection of OVA-IC. B6 mice were primed with 50 μg soluble OVA, 50 μg OVA incubated with 200 μg anti-HRP antibodies, or 50 μg OVA with 200 μg anti-OVA antibodies (OVA-IC). CD11c+ DCs were isolated 14 h after injection and analyzed for their ability to stimulate T cells in vitro. The indicated numbers of DCs were coincubated with RAG1-deficient OT-I cells (A) or purified CD4+ OT-II cells (B). Proliferation of cells was determined by [3H]thymidine incorporation. Error bars indicate SEM of triplicate wells.

Mentions: Previous in vitro studies have shown that immune complexes are much more efficiently taken up by DCs and cross-presented to T cells than soluble antigens (26, 27). To determine whether this enhancement also occurs in vivo, we injected B6 mice with 50 μg soluble OVA alone, OVA coincubated with a control anti-HRP IgG antibody, or OVA coincubated with anti-OVA IgG antibody (OVA-IC). 14 h after injection CD11c+ DCs were isolated from spleens and used as stimulators for naive MHC class I–restricted OVA-specific TCR transgenic OT-I, and MHC class II restricted OVA-specific TCR transgenic OT-II T cells in an in vitro proliferation assay. As shown by the data in Fig. 1 , DCs from mice injected with soluble OVA alone or together with the irrelevant antibody were not able to stimulate T cells. In contrast, injection of OVA-IC resulted in strong proliferation of OT-I and OT-II T cells, indicating an efficient uptake and presentation of OVA-IC by DCs in vivo.


Constitutive versus activation-dependent cross-presentation of immune complexes by CD8(+) and CD8(-) dendritic cells in vivo.

den Haan JM, Bevan MJ - J. Exp. Med. (2002)

DCs present OVA epitopes in association with both MHC class I and II after in vivo injection of OVA-IC. B6 mice were primed with 50 μg soluble OVA, 50 μg OVA incubated with 200 μg anti-HRP antibodies, or 50 μg OVA with 200 μg anti-OVA antibodies (OVA-IC). CD11c+ DCs were isolated 14 h after injection and analyzed for their ability to stimulate T cells in vitro. The indicated numbers of DCs were coincubated with RAG1-deficient OT-I cells (A) or purified CD4+ OT-II cells (B). Proliferation of cells was determined by [3H]thymidine incorporation. Error bars indicate SEM of triplicate wells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194052&req=5

fig1: DCs present OVA epitopes in association with both MHC class I and II after in vivo injection of OVA-IC. B6 mice were primed with 50 μg soluble OVA, 50 μg OVA incubated with 200 μg anti-HRP antibodies, or 50 μg OVA with 200 μg anti-OVA antibodies (OVA-IC). CD11c+ DCs were isolated 14 h after injection and analyzed for their ability to stimulate T cells in vitro. The indicated numbers of DCs were coincubated with RAG1-deficient OT-I cells (A) or purified CD4+ OT-II cells (B). Proliferation of cells was determined by [3H]thymidine incorporation. Error bars indicate SEM of triplicate wells.
Mentions: Previous in vitro studies have shown that immune complexes are much more efficiently taken up by DCs and cross-presented to T cells than soluble antigens (26, 27). To determine whether this enhancement also occurs in vivo, we injected B6 mice with 50 μg soluble OVA alone, OVA coincubated with a control anti-HRP IgG antibody, or OVA coincubated with anti-OVA IgG antibody (OVA-IC). 14 h after injection CD11c+ DCs were isolated from spleens and used as stimulators for naive MHC class I–restricted OVA-specific TCR transgenic OT-I, and MHC class II restricted OVA-specific TCR transgenic OT-II T cells in an in vitro proliferation assay. As shown by the data in Fig. 1 , DCs from mice injected with soluble OVA alone or together with the irrelevant antibody were not able to stimulate T cells. In contrast, injection of OVA-IC resulted in strong proliferation of OT-I and OT-II T cells, indicating an efficient uptake and presentation of OVA-IC by DCs in vivo.

Bottom Line: Murine splenic dendritic cells (DCs) can be divided into two subsets based on CD8alpha expression, but the specific role of each subset in stimulation of T cells is largely unknown.An important function of DCs is the ability to take up exogenous antigens and cross-present them in the context of major histocompatibility complex (MHC) class I molecules to CD8(+) T cells.These results suggest that although CD8(+) DCs constitutively cross-present exogenous antigens in the context of MHC class I molecules, CD8(-) DCs only do so after activation, such as via ligation of Fc(gamma)Rs.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195-7370, USA.

ABSTRACT
Murine splenic dendritic cells (DCs) can be divided into two subsets based on CD8alpha expression, but the specific role of each subset in stimulation of T cells is largely unknown. An important function of DCs is the ability to take up exogenous antigens and cross-present them in the context of major histocompatibility complex (MHC) class I molecules to CD8(+) T cells. We previously demonstrated that, when cell-associated ovalbumin (OVA) is injected into mice, only the CD8(+) DC subset cross-presents OVA in the context of MHC class I. In contrast to this selectivity with cell-associated antigen, we show here that both DC subsets isolated from mice injected with OVA/anti-OVA immune complexes (OVA-IC) cross-present OVA to CD8(+) T cells. The use of immunoglobulin G Fc receptor (Fc(gamma)R) common gamma-chain-deficient mice revealed that the cross-presentation by CD8(-) DCs depended on the expression of gamma-chain-containing activating FcgammaRs, whereas cross-presentation by CD8(+) DCs was not reduced in gamma-chain-deficient mice. These results suggest that although CD8(+) DCs constitutively cross-present exogenous antigens in the context of MHC class I molecules, CD8(-) DCs only do so after activation, such as via ligation of Fc(gamma)Rs. Cross-presentation of immune complexes may play an important role in autoimmune diseases and the therapeutic effect of antitumor antibodies.

Show MeSH
Related in: MedlinePlus