Composition, structural properties and immunomodulatory activity of several aqueous Pleurotus ß-glucan-rich extracts.

In this work, aqueous extracts from six different Pleurotus species were obtained and their yield, gross composition, ß-glucan content, monosaccharide profile, thermal stability, molecular weight distribution, and FT-IR were analyzed before and after purification through ethanol precipitation of the carbohydrate-rich fractions. The bioactivity (anti-inflammatory and immunomodulatory activity) of the various fractions obtained was also analyzed in three different cell cultures and compared with a lentinan control. The trend observed after purification of the aqueous fractions was an increase in the concentration of polysaccharides (especially ß-glucans), a decrease in ash, glucosamine and protein content and the elimination of low molecular weight (Mw) compounds, thus leaving in the purified samples high Mw populations with increased thermal stability. Interestingly, all these purified fractions displayed immunomodulatory capacity when tested in THP-1 macrophages and most of them also showed significant activity in HEK-hTLR4 cells, highlighting the bioactivity observed for Pleurotus ostreatus (both the extracts obtained from the whole mushroom and from the stipes). This specific species was richer in heteropolysaccharides, having moderate ß-glucan content and being enriched upon purification in a high Mw fraction with good thermal stability.

Polysaccharides from Pleurotus eryngii: Selective extraction methodologies and their modulatory effects on THP-1 macrophages.

Polysaccharides from P. eryngii mushroom were selectively extracted using low-cost technologies (water at different conditions of temperature and pressure). Mannogalactan was the main polysaccharide in cold-water extracted fraction (CWEF), while a linear (1→6)-ß-d-glucan was the main polymer in hot-water extracted fraction (HWEF). Autoclave-extracted fraction (AEF) contained a mixture of at least four different α- and ß-glucans. The report of linear (1→6)-ß-glucan and linear (1→3)-ß-glucan is a new finding for P. eryngii fruiting bodies. The immunostimulatory properties of the fractions on THP-1 macrophages were studied. All fractions at 50, 250 and 500 µg/mL were not cytotoxic and produced different stimulus on NO, IL-1ß and IL-10 secretion by the cells. Thus, our results showed that it is possible to concentrate different P. eryngii polysaccharides in selected fractions using a simple and low-cost procedure. Since biological effects depends on the polysaccharide structure, this technique allows the obtainment of fractions with distinct immunomodulatory activities.

Cationic lipid nanocarriers activate Toll-like receptor 2 and NLRP3 inflammasome pathways.

We provide evidence that cationic lipids, usually considered as a safe alternative to viral vectors as nanocarriers for gene therapy or drug intracellular delivery, do not behave as inert material but do activate cellular signalling pathways implicated in inflammatory reactions. We show here that the cationic lipid RPR206252 induces NF-κB activation, and the production of TNF-α, IL-1ß, IL-6 and IFN-γ by human or mouse macrophage cell lines. Further, we demonstrate that the activation of inflammatory cascades by RPR206252 is dependent on Toll-like receptor 2 (TLR2), the natural sensor of bacterial lipopeptides and NOD-like receptor protein 3 (NLRP3), the major inflammasome component. Our results suggest that cationic lipid nanocarriers because of their ability to stimulate the innate system can be used as a new class of synthetic and safe adjuvant for vaccination. FROM THE CLINICAL EDITOR: Cationic lipid nanocarriers are typically considered neutral tools for gene delivery. However, as demonstrated in this study, they possess a clear ability to stimulate the innate immune system, and actually can be used as a new class of synthetic and safe adjuvant for vaccination.

Effect of Echinacea purpurea (Asteraceae) aqueous extract on antibody response to Bothrops asper venom and immune cell response

The effect of aqueous extract of Echinacea purpurea roots on the murine antibody response to Bothrops asper snake venom in vivo was studied. Three groups were used. Group #1, baseline control, was treated with snake venom plus PBS. Group #2 was treated with snake venom plus sodium alginate as adjuvant (routine method used at Instituto Clodomiro Picado), and group #3 or experimental group, was treated with snake venom plus aqueous extract ofE. purpurea root as adjuvant. In all groups, the first inoculation was done with Freund's complete adjuvant (FCA). By the time of the second bleeding, mice in group #3 showed a remarkable increment in the level of anti-venom antibodies compared with those in groups #1 or #2. In vitro immune cell proliferation as a response to aqueous extract of E. purpurea root was studied using human lymphocytes activated with different lectins (Con A, PHA and PWM). In all cases, increase in percentage of lymphoproliferation was greater when E. purpurea root extract was used in addition to individual lectins.