Prothymosin α and a prothymosin α-derived peptide enhance TH1-type immune responses against defined HER-2/neu epitopes
1 Department of Animal and Human Physiology, Faculty of Biology, University of Athens, Athens 15784, Greece
2 Department of Internal Medicine II, Centre for Medical Research, University of Tübingen, Tübingen 72072, Germany
3 Department of Cell Biology and Biophysics, Faculty of Biology, University of Athens, Athens 15784, Greece
4 Interfakultäres Institut für Biochemie, University of Tübingen, Tübingen 72072, Germany
BMC Immunology 2013, 14:43 doi:10.1186/1471-2172-14-43Published: 22 September 2013
Additional file 1: Table S1:
Range of % cytokine positive CD4+ and CD8+ T cells. (A) Intracellular production of IFN-γ, TNF-α, IL-2, IL-4, IL-10 and IL-17 in, and expression of CD107 on CD4+ and CD8+ T cells stimulated with DCs matured with TNF-α, proTα or proTα(100–109), in the absence (−) or presence (+) of the HER-2/neu peptides. IL-5+ and IL-13+ CD8+ T cells are additionally shown. Numbers indicate percentages of positive cells. Shown is the range detected from 3–5 different donors tested. (B) Ratios of IFN-γ/IL-5 and IFN-γ/IL-13 in CD8+ T cells. Shown is the range from 3 different donors tested.
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Additional file 2: Figure S1:
Kinetics of CD14 and TLR-4 surface expression on monocytes, macrophages and iDCs/DCs upon stimulation with LPS, proTα or proTα(100–109). Monocytes, macrophages and iDCs (0 h) were stimulated with LPS (A), proTα (B), or proTα(100–109) (C) for 15 min, 30 min, 1 h and 18 h and assessed for the surface expression of CD14 and TLR-4 using flow cytometry. MFI values in the presence of neutralizing anti-TLR-4 Ab (+ a-TLR-4) are shown below each histogram. Histograms are from one representative donor of 3 tested. Using the loss of cell surface expression as a readout for TLR-4 and CD14 endocytosis from 0–36 h , data from all three donors are shown as mean values ± SDs for TLR-4 (D, E, F) and CD14 (G, H, I).
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Additional file 3: Figure S2:
CD14, TLR-4 and CD206 expression on monocytes, monocyte-derived macrophages and monocyte-derived iDCs. Macrophages were generated from human monocytes upon incubation with 100 ng/mL GM-CSF for 5 days. Human monocytes were isolated and iDCs were generated as described in Methods. Monocytes, macrophages and iDCs were assessed for the surface expression of CD14, TLR-4 and CD206 (as a specific marker for macrophages and DCs), using flow cytometry. Histograms are from one representative donor of 3 tested and numbers indicate MFIs.
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