Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2002 Sep 16;196(6):817-27.
doi: 10.1084/jem.20020295.

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

Joke M M den Haan  1 , Michael J Bevan
Affiliations
Comparative Study

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

Joke M M den Haan et al. J Exp Med. .

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.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
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.
Figure 2.
Figure 2.
Both CD8+ and CD8− DCs express FcγRII and RIII. CD11c+ DCs were purified from B6 (A–C), and from FcγRII-deficient (FcγRIIko; C), and mice deficient in all three FcγRs (FcγRko; C) and stained with antibodies specific for CD11c and CD8. DCs were gated on high expression of CD11c and the absence or presence of CD8. (A) FcγRI expression was determined by FACS® analysis of mouse IgG2a binding to CD8+ and CD8− DCs. Bold line depicts mouse IgG2a binding compared with background staining in the absence of mouse IgG2a. (B) FcγRII expression on CD8+ and CD8− DCs was determined by staining with specific anti-FcγRII antibody (bold line) compared with isotype control antibody (fine line). (C) The histograms show FcγRII/III expression of CD8+ and CD8− DCs from B6 mice (bold line), FcγRIIko (shaded histogram), and FcγRko mice (fine line).
Figure 3.
Figure 3.
Both CD8+ and CD8- DC subsets cross-present OVA-IC in association with MHC class I molecules, whereas presentation in association with MHC class II molecules is mainly restricted to CD8− DCs. B6 mice were injected with OVA-IC and 14 h after injection CD11c+ DCs were isolated. CD8+ and CD8− DC subsets were FACS® sorted and analyzed for their ability to stimulate T cells in vitro. 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.
Figure 4.
Figure 4.
Only CD8+ and not CD8− DCs from Fcγ-chain–deficient mice cross-present OVA-IC to CD8+ T cells. Fcγ-chain–deficient mice were injected with OVA-IC and 14 h after injection CD11c+ DCs were isolated. CD8+ and CD8− DC subsets were FACS® sorted and analyzed for their ability to stimulate T cells in vitro. Indicated numbers of DCs were coincubated with purified CD8+ 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.
Figure 5.
Figure 5.
CD8− DCs from FcγR-deficient mice have strongly decreased capacity to cross-present OVA-IC to CD8+ T cells. FcγR-deficient mice were injected with OVA-IC and 14 h after injection CD11c+ DCs were isolated. CD8+ and CD8− DC subsets were FACS® sorted and analyzed for their ability to stimulate T cells in vitro. Indicated numbers of DCs were coincubated with purified CD8+ 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.
Figure 6.
Figure 6.
Both DC subsets from B6 mice, Fcγ-chain–deficient mice and FcγR-deficient mice take up OVA-IC in vivo. CD11c+ DCs were isolated from control mice or mice injected with fluorescent DQ OVA-IC 14 h previously. Dot plots depict CD8 expression and uptake of DQ OVA by DCs isolated from B6 mice, Fcγ-chain–deficient mice and FcγR-deficient mice. DCs were gated on high CD11c expression, while autofluorescent cells were excluded.
Figure 7.
Figure 7.
Proliferation of OVA-specific CD8+ T cells in vivo after priming with OVA-IC is comparable in B6, Fcγ-chain–deficient mice and FcγR-deficient mice. CFSE-labeled, Thy1.1+ OT-I cells were transferred into B6, Fcγ-chain–deficient, and FcγR-deficient mice. 3 d later, mice were primed by injection of OVA-IC. Spleen cells were isolated 3 d after priming and stained for Thy1.1 and CD8. (A) CFSE profiles of Thy1.1+CD8+ OT-1 cells from mice that received no priming. (B) Same profiles from mice that were primed with OVA-IC.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Mellman, I., and R.M. Steinman. 2001. Dendritic cells: specialized and regulated antigen processing machines. Cell. 106:255–258. - PubMed
    1. Bevan, M.J. 1976. Minor H antigens introduced on H-2 different stimulating cells cross-react at the cytotoxic T cell level during in vivo priming. J. Immunol. 117:2233–2238. - PubMed
    1. Bevan, M.J. 1976. Cross-priming for a secondary cytotoxic response to minor H antigens with H-2 congenic cells which do not cross-react in the cytotoxic assay. J. Exp. Med. 143:1283–1288. - PMC - PubMed
    1. Huang, A.Y., P. Golumbek, M. Ahmadzadeh, E. Jaffee, D. Pardoll, and H. Levitsky. 1994. Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science. 264:961–965. - PubMed
    1. Kurts, C., W.R. Heath, F.R. Carbone, J. Allison, J.F. Miller, and H. Kosaka. 1996. Constitutive class I-restricted exogenous presentation of self antigens in vivo. J. Exp. Med. 184:923–930. - PMC - PubMed

Publication types