CD103⁻ dermal dendritic cells (dDCs) capture and transport glycosylated Tn antigen to the draining lymph nodes (DLN) for presentation to T cells. (A) Analysis of macrophage galactose-type lectin expression (bold line) in skin DLN DCs: CD8α⁺ DCs (CD11c⁺ major histocompatibility complex [MHC]) II⁺ CD11b⁻ CD8α⁺), Langerhans cells (CD11c⁺ MHCII⁺ CD8α⁻ Langerin⁺ CD103⁻), CD103⁺ dDC (CD11c⁺ MHCII⁺ CD8α⁻ Langerin⁺ CD103⁺), and CD103⁻ dDC (CD11c⁺ MHCII⁺ CD11b⁺ Langerin⁻ CD103⁻). Isotype control staining is shown as a gray histogram. (B-C) PBS, 10 nmol Alexa⁶⁴⁷-labeled glycosylated Tn antigen, or nonglycosylated control antigen was injected intradermally into the mouse ear. After 24 hours, we analyzed skin DLN, non-DLN, or spleen total CD11c⁺ DCs (B) and DLN DC subsets migrating from the skin (C) for antigen capture by flow cytometry. (D-E) We injected mice intradermally, in both ears, with 10 nmol multiple antigenic glycopeptide:Tn3-poliovirus (PV), multiple antigenic glycopeptide:Tn6-PV, nonglycosylated multiple antigenic peptides-PV peptide, or PBS. Total CD11c⁺ DCs (D) and DLN DC subsets migrating from the skin (E) were purified and cultured with a PV-specific T-cell hybridoma 24 hours later. IL-2 release by the T-cell hybridoma was analyzed in the 24-hour supernatants by enzyme-linked immunosorbent assay. Data are expressed as mean ± SD of duplicates. Representative results from 2 experiments are shown.