Macrophage Polarization and Receptor Activation
Corresponding Organization : Institut de Pharmacologie et de Biologie Structurale
Other organizations : Consejo Nacional de Investigaciones Científicas y Técnicas, Academia Nacional de Medicina, Centre for Genomic Regulation, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Inserm, Biomedical Primate Research Centre
Variable analysis
- Blocking antibodies specific to Dectin-1 (MAB1859, R&D Systems) or DC-SIGN (ab13847, abcam), or both
- Cytochalasin D (1 µg/ml, C8273, SIGMA) in combination with purified β-glucan from Saccharomyces cerevisiae (10 µg/ml, G5011, SIGMA)
- ManLAM (10 µg/ml, kindly provided by Dr. Jerome Nigou, IPBS/CNRS)
- LPS (1 µg/ml)
- Cytokine/chemokine production measured by ELISA analysis in the supernatants collected after 24 h
- Human macrophages differentiated at day 5 and activated with IL-4 (20 ng/ml) for additional 48 h
- M(IL-4) macrophages washed with warmed RPMI-1640 medium supplemented with M-CSF (10 ng/ml) and pre-incubated for 30 min
- M(IL-4) macrophages incubated with an irrelevant antibody (Ab-Control)
- Positive control: M(IL-4) macrophages treated with cytochalasin D (1 µg/ml) in combination with purified β-glucan from Saccharomyces cerevisiae (10 µg/ml) to efficiently stimulate Dectin-1
- Negative control: M(IL-4) macrophages incubated with an irrelevant antibody (Ab-Control)
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