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Spatial and mechanistic separation of cross-presentation and endogenous antigen presentation

Abstract

Antiviral or antitumor immunity requires activation of cytotoxic CD8+ T cells by dendritic cells, which present viral or tumor antigens on major histocompatibility complex (MHC) class I molecules. The intracellular mechanisms facilitating MHC class I–restricted presentation of extracellular antigens ('cross-presentation') are unclear. Here we demonstrate that cross-presentation of soluble antigen occurred in an early endosomal compartment distinct from the endoplasmic reticulum where endogenous antigen is loaded onto MHC class I. Efficient cross-presentation required endotoxin-induced, Toll-like receptor 4– and signaling molecule MyD88–dependent relocation of the transporter associated with antigen processing, essential for loading of MHC class I, to early endosomes. Transport of cross-presented antigen from endosomes to the cell surface was inhibited by primaquine, which blocks endosomal trafficking. Thus, cross-presentation is spatially and mechanistically separated from endogenous MHC class I–restricted antigen presentation and is biased toward antigens containing microbial molecular patterns.

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Figure 1: TAP is essential for cross-presentation and is present in OVA-containing endosomes.
Figure 2: US6 linked to Trf is specifically targeted into early endosomes.
Figure 3: Peptide loading for cross-presentation of soluble antigen occurs in early endosomes and not in the endoplasmic reticulum.
Figure 4: Cross-presented OVA is transported to the DC surface from endosomes, not from the endoplasmic reticulum.
Figure 5: Recruitment of TAP to early endosomes depends on endotoxin.
Figure 6: The endotoxin-mediated increase in cross-presentation depends on TLR4 and MyD88 and partially on TRIF.
Figure 7: Recruitment of TAP to endosomes depends on TLR4 and MyD88 but not on TRIF.

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Acknowledgements

We thank M.C. Nussenzweig (Rockefeller University) for mannose receptor-deficient mice; W.R. Heath (The Walter and Eliza Hall Institute of Medical Research) for OT-I and OT-II mice; R. Germain (National Institutes of Health) for 25-D1.16; A. Haas for the transferrin receptor antibody; and P. Knolle for critically reading the manuscript. We acknowledge the technical support of the flow cytometry core facility of the Institute of Molecular Medicine and Experimental Immunology and the House for Experimental Therapy of the Medical Faculty of Bonn University. Supported by the BONFOR program of Friedrich-Wilhelms University (O-173.0009 to S.B.) and by the Deutsche Forschungsgemeinschaft (SFB704 to C.K. and Ta157/7-1 to R.T.).

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S.B. designed experiments, did research, analyzed and interpreted data, and wrote the paper; C.S. and A.K. did experiments; R.T. contributed critical analytical tools and analyzed and interpreted data; C.K. designed experiments, analyzed and interpreted data, and wrote the paper.

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Correspondence to Sven Burgdorf or Christian Kurts.

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Burgdorf, S., Schölz, C., Kautz, A. et al. Spatial and mechanistic separation of cross-presentation and endogenous antigen presentation. Nat Immunol 9, 558–566 (2008). https://doi.org/10.1038/ni.1601

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