Cross-presenting CD103+ dendritic cells are protected from influenza virus infection
Julie Helft, … , Adolfo García-Sastre, Miriam Merad
Julie Helft, … , Adolfo García-Sastre, Miriam Merad
Published October 8, 2012
Citation Information: J Clin Invest. 2012;122(11):4037-4047. https://doi.org/10.1172/JCI60659.
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Research Article

Cross-presenting CD103+ dendritic cells are protected from influenza virus infection

Abstract

CD8+ cytotoxic T cells are critical for viral clearance from the lungs upon influenza virus infection. The contribution of antigen cross-presentation by DCs to the induction of anti-viral cytotoxic T cells remains controversial. Here, we used a recombinant influenza virus expressing a nonstructural 1–GFP (NS1-GFP) reporter gene to visualize the route of antigen presentation by lung DCs upon viral infection in mice. We found that lung CD103+ DCs were the only subset of cells that carried intact GFP protein to the draining LNs. Strikingly, lung migratory CD103+ DCs were not productively infected by influenza virus and thus were able to induce virus-specific CD8+ T cells through the cross-presentation of antigens from virally infected cells. We also observed that CD103+ DC resistance to infection correlates with an increased anti-viral state in these cells that is dependent on the expression of type I IFN receptor. These results show that efficient cross-priming by migratory lung DCs is coupled to the acquisition of an anti-viral status, which is dependent on the type I IFN signaling pathway.

Authors

Julie Helft, Balaji Manicassamy, Pierre Guermonprez, Daigo Hashimoto, Aymeric Silvin, Judith Agudo, Brian D. Brown, Mirco Schmolke, Jennifer C. Miller, Marylene Leboeuf, Kenneth M. Murphy, Adolfo García-Sastre, Miriam Merad

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Figure 3

CD103+ migratory DCs are not infected by influenza virus and uniquely preserve viral proteins in endosomal compartments.

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CD103+ migratory DCs are not infected by influenza virus and uniquely pr...
(A–I) Mice were infected with 106 PFUs of NS1-GFP virus. HA expression was measured by flow cytometry in CD45– lung cells (A) and alveolar macrophages (B) 15 hours after infection or in MLN CD103+ (C) and CD11b+ DCs (D) 48 hours after infection. Percentage of GFP+ cells in each population is noted in C and D. (E) Cell-surface HA expression was measured by confocal microscopy in MLN CD103+GFP+ DCs and CD11b+ DCs purified 48 hours after infection and lung alveolar macrophages purified 15 hours after infection. Original magnification, ×63, zoom 3. (F) Intracellular LAMP2 expression was analyzed by confocal microscopy in migratory CD103+GFP+ or CD11b+ DCs, isolated from the MLNs 48 hours after infection. Original magnification, ×63, zoom 3. (G) Intracellular NP expression was measured by flow cytometry in MLN cells isolated 48 hours after infection. Dot plots show percentage of intracellular NP expression among migratory CD103+ and CD11b+ DCs. (H and I) Affymetrix gene chip arrays of CD11b+ and CD103+ migratory DCs sorted from the MLNs of naive mice. Graphs represent mRNA transcript ratio between CD103+ DCs and CD11b+ DCs. Each dot represents 1 experiment. Bars represent the mRNA transcript absolute values for each gene. Data are representative of 3 separate experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.