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Blood, Vol. 108, Issue 5, 1643-1651, September 1, 2006
Dendritic cells and HIV-specific CD4+ T cells: HIV antigen presentation, T-cell activation, and viral transfer
Blood Moris et al.
108: 1643
Supplemental materials for: Moris et al
Files in this Data Supplement:
- Figure S1. Inhibition of MHC-II-restricted HIV antigen presentation by chloroquine (PDF, 40.5 KB) -
Primary immature DCs (A) or DC-SIGN+ B cells (B-BRE-DCS) (B) were preincubated for 30 min with chloroquine, an inhibitor of vesicular acidification, at the indicated concentrations. Cells were then exposed to HIVMN-AT2 (1 nM of p24) or gag1 peptide (22nM) for 2h at 37°C, fixed (0.5% PFA for 1min at 4°C) and co-cultivated with the HS clone IV9 for 8h. Activation of IV9 cells was assessed in an IFN Elipsot assay. Results are presented as the number of IFN+ positive cells for the indicated number of IV9 cells. Data are mean ± s.d. of triplicates and are representative of 3 independent experiments, with DCs from different donors.
- Figure S2. The control CD4+ T-cell clone 2.2 does not recognize HIV epitopes (PDF, 60.3 KB) -
ImDC were pulsed with HIVNL-AD8 (8nM of p24) or gag2 peptide (44nM) and co-cultivated with 2.2 cells for 6h. As positive controls for T cell activation, PHA (1 µg/ml), or PMA (50 ng/ml) with a calcium ionophore (Calcimycin, 1 µg/ml) were added in the co-cultures. Cells were then stained with anti-CD3, anti-DC-SIGN, and anti-cytokines (IL-2, TNF , and IFN) mAbs, and analyzed by flow cytometry. Results depicted were obtained by gating the analysis on 2.2 cells (CD3+, DCSIGN-). Figures indicate the percentage of cytokine+ cells. Similar results were obtained with 3.8 cells (not shown). Isotypic mAbs were used as negative controls to set the quadrant position. Data are representative of 3 independent experiments.
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