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. 2014 Aug 19;9(8):e103755. doi: 10.1371/journal.pone.0103755

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© 2014 Rauen et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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To study the effect of mannosylation on cross-presentation, we generated mannosylated and non-mannosylated SLPs containing the MHC I-restricted epitope of OVA (OVA_257–264; peptides 1 and 2). Mannosylation was introduced by a bis-mannosylated Lysin residue. Additionally, we generated mannosylated or non-mannosylated SLPs containing the MHC II-restricted epitope of OVA (OVA_323–339; peptides 3 and 4). To analyze intracellular trafficking, we synthesized fluorescently-labeled mannosylated or non-mannosylated SLPs (peptides 5 and 6). The Alexa647 fluorochrome was introduced by conjugation to an Alexa647-labeled cysteine. To study T cell responses against the E7 protein of HPV16, we generated mannosylated or non-mannosylated SLPs containing the MHC I restricted epitope of E7 (E7_43–63; peptides 7 and 8). Epitopes within the SLPs are in bold.