Galactomannan Downregulates the Inflammation Responses in Human Macrophages via NFκB2/p100

Figure 1

Galactomannan (GAL) attenuates the LPS-induced inflammation in human macrophages. (a) Schematic representation of the ET model used for this study. The cultures of human macrophages were or not pretreated with 10 ng/mL LPS or 10 μg/mL GAL for 24 h, washed twice with PBS, cultured in medium for 16 h, and restimulated with 10 ng/mL LPS for the indicated times. Supernatants were harvested at the indicated times (3 h, 5 h, and 24 h) and TNFα (b), IL6 (c), and IL10 (d) proteins levels were evaluated by CBA (); , comparing GAL + LPS with LPS. (e) Cell surface expression of HLA-DR was analyzed by flow cytometry. (), , comparing GAL  +  LPS with LPS 24 h. (f) Total mRNA was isolated from cells, cDNA was synthesized and CIITA real-time quantitative PCR was performed. (), comparing GAL + LPS with LPS 24 h. (g) Human heterologous lymphocytes were labeled with the membrane stain PKH2 Green Fluorescent Cell Linker Kit (Sigma-Aldrich) and cocultured with LPS, LPS + LPS, or GAL + LPS macrophages for 5 days. Lymphocyte proliferation was measured as a loss of green fluorescence intensity in the CD3⁺ gate, for this analysis. The fold induction is shown. , comparing GAL + LPS with LPS.

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