Mobilization of human CD34+ CD133+ and CD34+ CD133(-) stem cells in vivo by consumption of an extract from Aphanizomenon flos-aquae--related to modulation of CXCR4 expression by an L-selectin ligand?

OBJECTIVE: The goal of this study was to evaluate effects on human stem cells in vitro and in vivo of an extract from the edible cyanobacterium Aphanizomenon flos-aquae (AFA) enriched for a novel ligand for human CD62L (L-selectin). EXPERIMENTAL APPROACH: Ligands for CD62L provide a mechanism for stem cell mobilization in conjunction with down-regulation of the CXCR4 chemokine receptor for stromal derived factor 1. Affinity immunoprecipitation was used to identify a novel ligand for CD62L from a water extract from AFA. The effects of AFA water extract on CD62L binding and CXCR4 expression was tested in vitro using human bone marrow CD34+ cells and the two progenitor cell lines, KG1a and K562. A double-blind randomized crossover study involving 12 healthy subjects evaluated the effects of consumption on stem cell mobilization in vivo. RESULTS: An AFA extract rich in the CD62L ligand reduced the fucoidan-mediated externalization of the CXCR4 chemokine receptor on bone marrow CD34+ cells by 30% and the CD62L+ CD34+ cell line KG1A by 50% but did not alter the CXCR4 expression levels on the CD34(-) cell line K562. A transient, 18% increase in numbers of circulating CD34+ stem cells maximized 1 hour after consumption (P<.0003). When 3 noncompliant volunteers were removed from analysis, the increase in CD34+ cells was 25% (P<.0001). CONCLUSION: AFA water extract contains a novel ligand for CD62L. It modulates CXCR4 expression on CD34+ bone marrow cells in vitro and triggers the mobilization of CD34+ CD133+ and CD34+ CD133(-) cells in vivo.

Natural killer cell activation and modulation of chemokine receptor profile in vitro by an extract from the cyanophyta Aphanizomenon flos-aquae.

The present research was designed to study the effects of an extract from the edible cyanophyta Aphanizomenon flos-aquae on human natural killer (NK) cells. We have previously shown, using a double-blind randomized placebo-controlled crossover design, that ingestion of 1.5 g of dried whole A. flos-aquae resulted in a transient reduction in peripheral blood NK cells in 21 healthy human volunteers, suggesting increased NK cell homing into tissue. We have now identified an extract from A. flos-aquae (AFAe) that directly activates NK cells in vitro and modulates the chemokine receptor profile. NK cell activation was evaluated by expression of CD25 and CD69 on CD3-CD56+ cells after 18 hours. Changes in CXCR3 and CXCR4 chemokine receptor expression after 5-60 minutes were evaluated by immunostaining and flow cytometry. AFAe induced the expression of CD69 on CD3-CD56+ NK cells, induced CD25 expression on 25% of these cells, and acted in synergy with interleukin 2. NK cells enriched by RosetteSep (StemCell Technologies Inc., Vancouver, BC, Canada) were not activated by AFAe, indicating that the NK activation was dependent on other cells such as monocytes. The low-molecular-weight fraction <5,000 of AFAe was responsible for the most robust NK cell activation, suggesting novel compounds different from previously reported macrophage-activating large polysaccharides.

An antiinflammatory immunogen from yeast culture induces activation and alters chemokine receptor expression on human natural killer cells and B lymphocytes in vitro.

The aim of this study was to evaluate the immunomodulating effects of a consumable yeast-based immunogen, EpiCor, on human leukocytes in vitro. The selection of antiinflammatory and lymphocyte activation assays was based on initial evidence for immunomodulating effects of EpiCor from an unusually low incidence of influenza among employees in a factory manufacturing EpiCor, along with a high oxygen radical absorbance capacity value. In the present study, EpiCor significantly reduced the production of reactive oxygen species by neutrophils (P < .005). EpiCor treatment of peripheral blood mononuclear cells (PBMCs) caused induction of the activation markers CD80 and CD86 on B lymphocytes, and CD69 and CD25 on CD3-CD56+ natural killer cells. This induction was also seen on enriched populations of natural killer and B lymphocytes, suggesting a direct effect not dependent on bystander cells. Coculturing of PBMC with EpiCor and phytohemagglutinin resulted in inhibition of phytohemagglutinin-induced T-cell proliferation and reduction of interferon gamma production. Fucoidan, a ligand for the homing molecule l-selectin (CD62L), is known to induce rapid up-regulation of several chemokine receptors on lymphocytes. EpiCor caused strong inhibition of Fucoidan-mediated expression of the chemokine receptors CXCR4 and CCR9 on PBMC. This suggested rapid altering of signal transduction pathways, or a direct competition for cell surface receptors, with an end result being an altered sensitivity to chemotactic signals from tissue. We conclude that EpiCor possesses significant antiinflammatory activity and induces direct activation and increased chemotactic awareness of lymphocyte subsets in vitro. This suggests further study of effects of EpiCor consumption on antiviral defense mechanisms and antibody production.

Antioxidant capacity and other bioactivities of the freeze-dried Amazonian palm berry, Euterpe oleraceae mart. (acai).

The fruit of Euterpe oleraceae, commonly known as acai, has been demonstrated to exhibit significantly high antioxidant capacity in vitro, especially for superoxide and peroxyl scavenging, and, therefore, may have possible health benefits. In this study, the antioxidant capacities of freeze-dried acai fruit pulp/skin powder (OptiAcai) were evaluated by different assays with various free radical sources. It was found to have exceptional activity against superoxide in the superoxide scavenging (SOD) assay, the highest of any food reported to date against the peroxyl radical as measured by the oxygen radical absorbance capacity assay with fluorescein as the fluorescent probe (ORACFL), and mild activity against both the peroxynitrite and hydroxyl radical by the peroxynitrite averting capacity (NORAC) and hydroxyl radical averting capacity (HORAC) assays, respectively. The SOD of acai was 1614 units/g, an extremely high scavenging capacity for O2*-, by far the highest of any fruit or vegetable tested to date. Total phenolics were also tested as comparison. In the total antioxidant (TAO) assay, antioxidants in acai were differentiated into "slow-acting" and "fast-acting" components. An assay measuring inhibition of reactive oxygen species (ROS) formation in freshly purified human neutrophils showed that antioxidants in acai are able to enter human cells in a fully functional form and to perform an oxygen quenching function at very low doses. Furthermore, other bioactivities related to anti-inflammation and immune functions were also investigated. Acai was found to be a potential cyclooxygenase (COX)-1 and COX-2 inhibitor. It also showed a weak effect on lipopolysaccharide (LPS)-induced nitric oxide but no effect on either lymphocyte proliferation and phagocytic capacity.

Immunomodulation by SanPharma fungal metabolic products.

OBJECTIVE: In the present study, a series of fungal metabolite products from SanPharma (Dohren, Germany) were tested for effects on human peripheral blood leukocytes in vitro using standard immunologic methods. BACKGROUND: Therapeutic strategies used in German biological medicine often include treatment (oral, nasal, rectal, topical, or injection) with fungal or bacterial products, also known as "isopathic remedies," of which some are limited to metabolic products, whereas others include microbial cell lysates and cell wall fragments as well. The SanPharma products are based on metabolites, and do not contain microbial cell wall compounds. METHODS: Activation of natural killer (NK) cells was evaluated by cell surface immunostaining using CD3, CD56, CD69, and CD25 monoclonal antibodies. Production of interferon-gamma was evaluated by enzyme-linked immunoabsorbent assay (ELISA) on supernatants collected after 5 days' culture of peripheral blood mononuclear cells (PBMC). Direct mitogenic effect was assessed using the lipophilic membrane dye PHK26 (Sigma-Aldrich, St. Louis, MO) in a fluorescence-based proliferation assay, in which fluorescence intensity is reduced upon cell divisions. Cell viability upon exposure to fungal metabolites was assessed using propidium iodide staining and flow cytometry. RESULTS: All fungal metabolite products specifically induced the expression of the CD69 marker on human CD3-negative, CD56-positive NK cells, but not CD3-positive T cells, in vitro, as shown by the induction of the CD69 marker on up to 50% of NK cells after 18 hours' culture with metabolites. Only one of the five metabolite products, Roqueforti, induced cyclooxygenase-2 (COX-2), indicating that nuclear factor-kappaB (NFkappaB)-mediated signaling may not have been involved in the NK activation by the other four products. The Notatum product reduced baseline levels of COX-2, indicating an anti-inflammatory effect. No evidence of toxic or mitogenic effects was found. CONCLUSIONS: The fungal metabolite products from SanPharma specifically activate human NK cells in vitro.