Polysaccharides of Ganoderma lucidum alter cell immunophenotypic expression and enhance CD56+ NK-cell cytotoxicity in cord blood.

In our previous study, a fucose-containing glycoprotein fraction (F3), isolated from the water-soluble extracts of Ganoderma lucidum, was shown to stimulate mice spleen cell proliferation and cytokine expression. We now further investigate the effect of F3 on the immunophenotypic expression in mononuclear cells (MNCs). When human umbilical cord blood (hUCB) MNCs were treated with F3 (10-100 microg/mL) for 7days, the population of CD14+CD26+ monocyte/macrophage, CD83+CD1a+ dendritic cells, and CD16+CD56+ NK-cells were 2.9, 2.3, and 1.5 times higher than those of the untreated controls (p<0.05). B-cell population has no significant change. T cell growth was, however, slightly inhibited and CD3 marker expression decreased approximately 20% in the presence of higher concentrations of F3 (100 microg/mL). We also found that F3 is not harmful to human cells in vitro, and after F3 treatment, NK-cell-mediated cytotoxicity was significantly enhanced by 31.7% (p<0.01) at effector/target cell ratio (E/T) 20:1, but was not altered at E/T 5:1.

Cell phenotype analysis using a cell fluid-based microchip with high sensitivity and accurate quantitation.

We have assessed a cell fluid chip-based fluorescent cytometric assay that runs on bioanalyzer for fast characterization of small population cell phenotypes characterization. The assay determines the expression of specific cell surface markers on various cell samples. Six samples can be analyzed on each chip in one automated process. Results were in good agreement with conventional flow cytometry in quantitation. Importantly, this procedure used less than 200 cells per sample and produced results consistent with that using 10(5) cells by the conventional staining procedure. The method was also used for screening potential ingredients in herbs. Purpose of this study was to analyze the change of cell subtypes of UCB mononuclear cells in vitro reactivity in herbs. We found that by treatment of the water-soluble extract (F3) of Ganoderma lucidum, the presence of CD56(+) marker (natural killer cells) significantly increased from 1.1 to 3.2% (P<0.05 and P) in UCB mononuclear cells. The results indicated that F3 quantitatively influenced NK cells activities. We suggest this screening method may be useful for a fast phenotypes characterization after extract stimulation utilizing only a small population of cells.