The DC receptor DNGR-1 mediates cross-priming of CTLs during vaccinia virus infection in mice
Salvador Iborra, … , Caetano Reis e Sousa, David Sancho
Salvador Iborra, … , Caetano Reis e Sousa, David Sancho
Published April 16, 2012
Citation Information: J Clin Invest. 2012;122(5):1628-1643. https://doi.org/10.1172/JCI60660.
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Research Article

The DC receptor DNGR-1 mediates cross-priming of CTLs during vaccinia virus infection in mice

Abstract

In order to prime T cells, DCs integrate signals emanating directly from pathogens and from their noxious action on the host. DNGR-1 (CLEC9A) is a DC-restricted receptor that detects dead cells. Therefore, we investigated the possibility that DNGR-1 affects immunity to cytopathic viruses. DNGR-1 was essential for cross-presentation of dying vaccinia virus–infected (VACV-infected) cells to CD8+ T cells in vitro. Following injection of VACV or VACV-infected cells into mice, DNGR-1 detected the ligand in dying infected cells and mediated cross-priming of anti-VACV CD8+ T cells. Loss of DNGR-1 impaired the CD8+ cytotoxic response to VACV, especially against those virus strains that are most dependent on cross-presentation. The decrease in total anti-VACV CTL activity was associated with a profound increase in viral load and delayed resolution of the primary lesion. In addition, lack of DNGR-1 markedly diminished protection from infection induced by vaccination with the modified vaccinia Ankara (MVA) strain. DNGR-1 thus contributes to anti-VACV immunity, following both primary infection and vaccination. The non-redundant ability of DNGR-1 to regulate cross-presentation of viral antigens suggests that this form of regulation of antiviral immunity could be exploited for vaccination.

Authors

Salvador Iborra, Helena M. Izquierdo, María Martínez-López, Noelia Blanco-Menéndez, Caetano Reis e Sousa, David Sancho

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Figure 5

DNGR-1 expression on transferred CD8α+ spleen DCs is crucial for their ability to cross-present VACV antigens.

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DNGR-1 expression on transferred CD8α+ spleen DCs is crucial for their a...
(A) Splenocytes from WT or Clec9agfp/gfp mice were extracted and cultured for 2 hours with RAW cells infected with WR VACV and irradiated with UV to inactivate the virus (RAW-VACV-UV). CD8α+ DCs were purified as indicated in Methods and transferred into the hind paws of Clec9agfp/gfp mice (2 × 106 CD8α+ DCs per mouse). (B) WT but not DNGR-1–deficient CD8α+ DCs cross-present VACV-derived antigens. After 7 days, splenocytes were extracted and restimulated with B8R or A3L VACV peptides. Production of IFN-γ is shown as individual data from a representative experiment (n = 7 biological replicates) of 3 independent experiments performed. (C and D) Cross-presentation of VACV antigens by WT but not DNGR-1–deficient CD8α+ DCs results in CTL killing activity against vaccinia epitopes in vivo. On day 6 after transfer, splenocytes from syngeneic mice were loaded with the early peptide B8R, the late peptide A3L, or no peptide and labeled as in Figure 4 and transferred i.v. Splenocytes were analyzed 16 hours later for specific killing of targets. (C) Representative histogram set. DCs transferred without preincubation with RAW-VACV-UV show the proportion of transferred targets. (D) Percentage specific killing shown as individual data from a representative experiment (n = 4 biological replicates) of 3 performed. #P < 0.001, unpaired 2-tailed Student’s t test.