Journal:

Article Title: Metastatic Melanoma Secreted IL-10 Down-Regulates CD1 Molecules on Dendritic Cells in Metastatic Tumor Lesions

doi:

Figure Lengend Snippet: IL-10 down-regulates CD1 expression on monocyte-derived DC in a dose-dependent manner. Monocyte-derived DC were cultured for 3 days in the presence of IL-4 (1000 IU/ml) and GM-CSF (800 IU/ml). Monocyte-derived DC were thereafter cultured for another 72 hours in GM-CSF and IL-4 in the presence of various IL-10 concentrations (1, 10, and 100 ng/ml). A: Analysis of CD1a, b, c and CD86, CD80, and (B) HLA-DR expression by flow cytometry. Monocyte-derived DC cultured in presence of GM-CSF and IL-4 without IL-10 were used as positive control. Results are representative for three independent experiments. Filled histograms represent isotype controls.

Article Snippet: In some experiments DC were also stained with the following PE or FITC-conjugated monoclonal antibodies: anti-CD11a (IgG 2a ), anti-CD11b (Ig G1 ), anti-CD11c (Ig G1 ), anti-CD14 (IgG 2a ), anti-CD80 (Ig G1 ), anti-CD83 (Ig G1 ), and anti-CD86 (Ig G1 ) (all from Becton Dickinson).

Techniques: Expressing, Derivative Assay, Cell Culture, Flow Cytometry, Positive Control

Journal:

Article Title: Metastatic Melanoma Secreted IL-10 Down-Regulates CD1 Molecules on Dendritic Cells in Metastatic Tumor Lesions

doi:

Figure Lengend Snippet: Effects of melanoma supernatants on CD1, CD86, and HLA-DR expression on monocyte-derived DC. A: Supernatants from first passage metastatic melanoma cultures with high (cell line 000630, 306 pg/ml IL-10) and low (cell line 000515, 91 pg/ml IL-10) IL-10 concentrations were used to study the effects of different supernatant dilutions on CD1 expression on GM-CSF/IL-4-generated DC. After 3 days, 0%, 10%, 50%, and 100% of the medium was replaced with melanoma supernatants from the melanoma cell culture indicated and incubated for another 3 days. CD1a, b, and c expression was thereafter assessed by flow cytometry. B: CD86, CD80, and HLA-DR expression on DC after replacement of 0%, 10%, 50%, and 100% of DC culture medium with melanoma supernatant (cell line 000630). C: Supernatants from different melanoma and fibroblast cell cultures were used to study the effects on CD1 expression on DC using flow cytometry. Fifty percent of the DC culture medium was replaced with first passage metastatic melanoma (culture 000630), second passage metastatic melanoma (culture 000621), primary melanoma (culture 990220), and fibroblast cell culture supernatants. CD1 expression was thereafter measured using flow cytometry. Results are shown as the mean MFI of three independent experiments. Vertical bars represent the standard deviations of the means. *, P < 0.05, in relation to monocyte-derived DC cultured in the absence of cell culture supernatant. MM, metastatic melanoma; PM, primary melanoma; fibro, fibroblasts; P, passage.

Article Snippet: In some experiments DC were also stained with the following PE or FITC-conjugated monoclonal antibodies: anti-CD11a (IgG 2a ), anti-CD11b (Ig G1 ), anti-CD11c (Ig G1 ), anti-CD14 (IgG 2a ), anti-CD80 (Ig G1 ), anti-CD83 (Ig G1 ), and anti-CD86 (Ig G1 ) (all from Becton Dickinson).

Techniques: Expressing, Derivative Assay, Generated, Cell Culture, Incubation, Flow Cytometry