The dendritic cell receptor DNGR-1 controls endocytic handling of necrotic cell antigens to favor cross-priming of CTLs in virus-infected mice
Santiago Zelenay, Anna M. Keller, Paul G. Whitney, Barbara U. Schraml, Safia Deddouche, Neil C. Rogers, Oliver Schulz, David Sancho, Caetano Reis e Sousa
Santiago Zelenay, Anna M. Keller, Paul G. Whitney, Barbara U. Schraml, Safia Deddouche, Neil C. Rogers, Oliver Schulz, David Sancho, Caetano Reis e Sousa
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Research Article

The dendritic cell receptor DNGR-1 controls endocytic handling of necrotic cell antigens to favor cross-priming of CTLs in virus-infected mice

Abstract

DNGR-1 (CLEC9A) is a receptor for necrotic cells required by DCs to cross-prime CTLs against dead cell antigens in mice. It is currently unknown how DNGR-1 couples dead cell recognition to cross-priming. Here we found that DNGR-1 did not mediate DC activation by dead cells but rather diverted necrotic cell cargo into a recycling endosomal compartment, favoring cross-presentation to CD8+ T cells. DNGR-1 regulated cross-priming in non-infectious settings such as immunization with antigen-bearing dead cells, as well as in highly immunogenic situations such as infection with herpes simplex virus type 1. Together, these results suggest that DNGR-1 is a dedicated receptor for cross-presentation of cell-associated antigens. Our work thus underscores the importance of cross-priming in immunity and indicates that antigenicity and adjuvanticity can be decoded by distinct innate immune receptors. The identification of specialized receptors that regulate antigenicity of virus-infected cells reveals determinants of antiviral immunity that might underlie the human response to infection and vaccination.

Authors

Santiago Zelenay, Anna M. Keller, Paul G. Whitney, Barbara U. Schraml, Safia Deddouche, Neil C. Rogers, Oliver Schulz, David Sancho, Caetano Reis e Sousa

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Figure 2

DNGR-1 does not behave as a myeloid activatory receptor.

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DNGR-1 does not behave as a myeloid activatory receptor. BM cells from D...
BM cells from Dectin-1–deficient mice were retrovirally transduced with a vector encoding for Dectin-1 or for a chimeric receptor (Chimera) bearing the cytoplasmic tail of DNGR-1 (WT) or 3 mutant versions thereof (Y7F, A3D, and I6G) fused to the transmembrane, stalk region, and CTLD of Dectin-1. All constructs were followed by an IRES-GFP sequence, which allowed sorting based on GFP levels to normalize for Dectin-1 or chimera surface expression. (A) Schematic representation of the chimeric DNGR-1/Dectin-1 receptors. Asterisks denote the mutated residues. (B) Expression of Dectin-1 or the different chimeras by GFP+ GMCSF-BMDCs purified by cell sorting as determined by staining with an anti–Dectin-1 antibody before overnight stimulation. (C) Purified GFP+ DCs as in B were cultured overnight in the presence of increasing amounts of curdlan (0, 30, and 100 μg/ml). Concentrations of TNF-α, IL-2, and IL-10 in the supernatant after culture are shown. Results are mean ± sem and representative of at least 3 independent experiments. ND, not detected.