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 4

DNGR-1 diverts cargo to the early/recycling endosomal pool.

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DNGR-1 diverts cargo to the early/recycling endosomal pool. (A) Purified...
(A) Purified splenic CD8α+ DCs from WT mice were cultured for 30 minutes (EEA-1) or for 90 minutes (Rab5a and Rab27a) with Alexa Fluor 488–coupled anti–DNGR-1 Ab. Cells were fixed and stained for confocal microscopy. Distribution of internalized anti–DNGR-1 Ab+ vesicles relative to EEA-1+, Rab5a+, or Rab27a+ vesicles was quantified as percentage of vesicles positive for anti–DNGR-1 antibody also positive for endosomal markers in 15 cells per condition. (B and C) Purified CD8α+-like Flt3L-BMDCs from WT or DNGR-1–deficient mice were cultured with UV-treated Alexa Fluor 647–labeled H-2bm1 MEFs. Cells were fixed after 60 or 240 minutes and stained for confocal microscopy. Distribution of cargo derived from dead cells relative to Rab5a (B) and Rab11 (C) was quantified as percentage of total vesicles positive for endosomal markers containing dead cell cargo in 15 cells per condition. Scale bars: 5 μm. Results are representative of 2 independent experiments. *P < 0.05, **P < 0.01, unpaired Student’s t test.