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eji2283-sup-0001-s1.pdf554KFigure S1. CD141+ DCs are functional despite being unable to phagocytose E. coli. A. CD141+ DCs fail to take up E. coli (upper left plot, grey: control, white: E. coli-GFP). The same batch of CD141+ DCs take up FITC-dextran (upper right plot, grey: control, white: FITC-dextran) after 1h at 37oC. CD141+ DCs from the same isolation also upregulate MHC class I (lower left plot) and MHC class II (lower right plot) after overnight culture (black line open) and in response to E. coli (black filled). Grey filled histogram is freshly isolated DCs and grey filled histogram is isotype control. One of 2 similar experiments is shown. B. CD141+ DCs produce low levels of IL-8 after overnight culture and upregulate IL-8 after exposure to E. coli. One of three representative donors is shown. C. The presence of E. coli does not impair direct peptide presentation by any of the DC subsets. DCs were pulsed with the HLA-A*0201-restricted FMP58-66 peptide or an irrelevant HLA-A*0021- in the presence or absence of E. coli for 1h then washed and presentation to a FMP58-66 specific CD8+ T cell clone was measured by specific IFN-γ production. One representative of two experiments is shown. D. CD141+ DCs from the same isolation can directly present the FMP58-66 epitope when loaded with peptide but not E. coli/LLO expressing FMP protein. Controls were an irrelevant peptide, or E. coli/LLO that doesn't express FMP. One of three representative experiments is shown.
eji2283-sup-0001-s1.pdf554KFigure S2. CD1c+ DCs produce IL-10 in response to different stimuli. Secretion of IL-10 by MoDCs and CD1c+ DCs per 5x104 cells from the same donor in response to no activation (no act) or activation with polyI:C (10μg/ml), LPS (100ng/ml) or E. coli after 24 hours was analysed by flow cytometric bead arrays. One representative of 3 donors is shown.
eji2283-sup-0001-s1.pdf554KFigure S3. Surface molecular expression on CD1c+ DCs and MoDCs activated with E. coli. MoDCs or CD1c+ DCs of the same donor were cultured with or without E. coli for 24 h, and the surface expression of CD40, CD83, CD86 and ICOS-L were analysed by flow cytometry. Numerical values are mean fluorescence intensity after deduction of the value from an isotype control (grey). One representative experiment of three is shown.

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