polyclonal goat anti-human cxcl16 (purified biotin labeled  (R&D Systems)

Journal: Mediators of Inflammation

Article Title: Plasmacytoid Dendritic Cell Response to CpG ODN Correlates with CXCL16 Expression and Is Inhibited by ox-LDL

doi: 10.1155/2013/312590

Figure Lengend Snippet: Metalloproteinase inhibitor GM-6001 enhances cytokine production induced by D-ODN. (a) PBMC (4 × 10 6 /mL) were preincubated in the absence or presence of GM6001 (50 μ M) for 30 min, fixed, and stained for CXCL16 expression. Cells treated as in (a) were washed and then stimulated with 3 μ M D-ODN (b), 1 μ M of C ODN (c), or 1 μ M of K-ODN (d) for 24 h. Cytokine production (IL-6 for K-ODN and IFN α for D and C ODN) was assessed from culture supernatants using ELISA. Response of 6 individual donors is shown.

Article Snippet: Polyclonal goat anti-human CXCL16 (purified and biotin labeled) and its isotype matched control were from R&D Systems.

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Plasmacytoid Dendritic Cell Response to CpG ODN Correlates with CXCL16 Expression and Is Inhibited by ox-LDL

doi: 10.1155/2013/312590

Figure Lengend Snippet: Altering CXCL16 expression influences “D” ODN induced cytokine production.

Article Snippet: Polyclonal goat anti-human CXCL16 (purified and biotin labeled) and its isotype matched control were from R&D Systems.

Techniques: Expressing

Journal: Mediators of Inflammation

Article Title: Plasmacytoid Dendritic Cell Response to CpG ODN Correlates with CXCL16 Expression and Is Inhibited by ox-LDL

doi: 10.1155/2013/312590

Figure Lengend Snippet: Digestion of CXCL16 on the surface of pDCs reduces cytokine production induced by D-ODN. (a) PBMC (4 × 10 6 /mL) were preincubated in the absence or presence of O-sialoglycoprotein endopeptidase (25 μ g/mL) for 30 min, fixed, and stained for CXCL16 expression. (b) Cells treated as in (a) were washed and then stimulated with 3 μ M D-ODN for 24 h. Cytokine production was assessed from culture supernatants using ELISA. Response of 6 individual donors is shown.

Article Snippet: Polyclonal goat anti-human CXCL16 (purified and biotin labeled) and its isotype matched control were from R&D Systems.

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Plasmacytoid Dendritic Cell Response to CpG ODN Correlates with CXCL16 Expression and Is Inhibited by ox-LDL

doi: 10.1155/2013/312590

Figure Lengend Snippet: CXCL16/CXCR6 interaction reduces the threshold of activation in pDCs responding to D-ODN. (a) pDCs were enriched from human PBMCs using the BDCA-4 isolation kit as recommended by the manufacturer. Purity of cells was established by flow cytometry following staining with pDC associated cell-surface markers. (b) Enriched pDCs (100,000 cells/well) were incubated with CXCR6 or CCR5 expressing Jurkat T cells (1 : 1 ratio) in 96-well U-bottom plates. Cocultures were then stimulated with a suboptimal concentration of D-ODN (0.75 μ g/mL). pDC stimulated with optimal concentration of D-ODN (10 μ g/mL) served as a positive control. Cytokine production was assessed from culture supernatants 24 h later using ELISA. Average response of 3 different pDC preparations is shown (mean ± S.D).

Article Snippet: Polyclonal goat anti-human CXCL16 (purified and biotin labeled) and its isotype matched control were from R&D Systems.

Techniques: Activation Assay, Isolation, Flow Cytometry, Staining, Incubation, Expressing, Concentration Assay, Positive Control, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Plasmacytoid Dendritic Cell Response to CpG ODN Correlates with CXCL16 Expression and Is Inhibited by ox-LDL

doi: 10.1155/2013/312590

Figure Lengend Snippet: Oxidized LDL and recombinant IFN γ /TNF α modify cytokine production induced by D-ODN. (a) Enriched pDCs (100,000 cells/well) were preincubated with LDL or OxLDL (10 μ g/mL each) and then stimulated with 3 μ M of D-ODN. Cytokine production was assessed from culture supernatants 24 h later using ELISA. Average response of 3 different pDC preparations is shown (mean ± S.D). (b) Enriched pDCs were preincubated in the absence or presence of GM6001 (50 μ M), recIFN γ /TNF α (20 ng/mL each), or PMA/ionomycin (250 pg/mL/100 pg/mL) for 30 min at 37°C. Cytokine production was assessed from culture supernatants 24 h later using ELISA. Average response of 3 different pDC preparations is shown (mean ± S.D). (c) PBMC (4 × 10 6 /mL) were preincubated in the absence or presence of 1 μ M Control ODN, 1 μ M K-ODN, or 3 μ M D-ODN for 48 h. Cells were then fixed and stained for CXCL16 expression. MFI of CXCL16 stained cells ± S.D of 3 different donors is shown.

Article Snippet: Polyclonal goat anti-human CXCL16 (purified and biotin labeled) and its isotype matched control were from R&D Systems.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Control, Staining, Expressing