Clinical Studies

IMMUNE PARAMETERS STUDY – CELT NATURALS IMMUNO-CARE^®
CONDUCTED AT THE UNIVERSITY OF GUELPH, BY THE HUMAN NUTRACEUTICAL RESEARCH UNIT

The following randomized, double-blind, placebo-controlled clinical trial involving 20 subjects over 28 days, demonstrated that Immuno-Care^®, by Celt Naturals, Calgary, Canada, can have a beneficial effect on immune parameters. The clinical trial showed a reduction in basophils that release histamine, typically found in allergic reactions. We are pleased to provide the findings of this important trial and encourage you to contact Celt Naturals.

The Supplement Immuno-Care^® by Celt Naturals

Human Nutraceutical Research Unit, J.T. Powell Building. University of Guelph, Guelph,
Ontario, Canada N1G 2W1, Tel: (519) 824-4120, Ext. 5374, Fax: (519) 823-5247
Website: www.uoguelph.ca/hnru, Maggie Laidlaw, M.Sc., Interim Director, HNRU.

OBJECTIVE:

This trial was to determine the effects of the supplement Immuno-Care^®, by Celt Naturals, Calgary, Canada, containing plant sterols, pine bark antioxidants, and an essential fatty acid complex, on specific immune parameters and cardiovascular indices. The trial was conducted at the University of Guelph as a randomized, double-blind, placebo-controlled clinical trial, with 20 subjects over 28 days.

ABSTRACT

A randomized, double-blind, placebo-controlled clinical trial to determine the effects of Celt Naturals Immuno-Care^® supplement (containing plant sterols, pine bark antioxidants and essential fatty acid complex) on specific immune parameters and cardiovascular indices in both men and women with non-food allergies.

The supplement appears to have effect on immune parameters, particularly basophils and Il-6 levels. Given these results, this supplement would appear to have the potential to substantially alleviate allergic responses.

The supplement also significantly reduced circulating levels of LDL cholesterol and increased circulating levels of HDL cholesterol. There was a significant decrease in the ratio of TC/HDL, and in the ratio of LDL/HDL cholesterol, which corresponds to a decrease in cardiovascular risk, as these ratios are markers for a reduction in the risk for developing atherosclerosis.

This information is provided for educational purposes only and should not be construed as medical advice. Any reader requiring medical advice is strongly advised to consult a licensed healthcare professional. Any products mentioned are not intended to diagnose, treat, cure, mitigate, or prevent any disease. These statements have not been evaluated by the United States Food and Drug Administration (USFDA).

RESULTS

Immune Parameters

The effects of the supplement on immune parameters are presented in Table 1. A number of studies support the belief that human basophils play an important role in allergic inflammation. Mast cells and basophils express the high affinity receptor for IgE (FcepsilonRI) and play a central role for IgE-associated immediate hypersensitivity reactions and allergic disorders. During allergic reactions, basophils migrate from the blood compartment to inflammatory sites, where they act as effector cells, in concert with eosinophils. Basophils release histamine during inflammation and allergic reactions.

Table 1:

**Statistically significant, p<0.0.

The participants in the treatment group, when compared to the control group, showed a significant reduction in basophil count, while the reduction seen in the control group was non-significant.
A reduction in basophil count may indicate a reduction in histamine release.

The immune system also responds to stressors by causing certain immune cells to secrete the
pro-inflammatory cytokines, Interleukin-1 (IL-1) and Interleukin-6 (IL-6). These cytokines are both involved in inflammation, and IL-6, in particular, is thought to worsen the symptoms of autoimmune diseases and fibromyalgia.

IL-6 has been found to act as a growth factor in several tumors, and some viruses also use IL-6 to replicate. IL-6 also causes calcium to be released from bone, promoting osteoporosis. We must control the release of these cytokines if we want to enhance immunity and reduce degenerative diseases.

It was noted in the pilot trial that the pro-inflammatory cytokine IL-6 levels showed a substantial reduction in the treated group, when compared to the control group. Although the drop in the
IL-6 levels in the treatment group was not statistically significant, a larger study with more subjects, over a longer period of time, may show significance.

Celt Naturals Immuno-Care^® has demonstrated that it has an effect on histamine-containing basophil counts, with a reduction of IL-6 levels, and consequently may substantially alleviate symptoms associated with airborne allergens, asthma, and allergic rhinitis. Further studies are recommended, with a larger patient participation and a longer trial period, to investigate other areas of immunological response.

Cardiovascular Parameters

The effects of the supplement on lipid and lipoprotein parameters and cardiovascular indices are illustrated in Tables 2 and 3.

Table 2: The effects of Celt Immuno-Care^® on blood lipid parameters in experimental and placebo groups from Day 0 to Day 28.

**Statistically significant, p<0.05.

Table 3: The effects of Celt Immuno-Care^® supplementation on specific cardiovascular ratios in experimental and placebo groups from Day 0 to Day 28.

**Statistically significant, p<0.05.

The specific objective of this portion of the trial was to determine the effects of the supplement Celt Immuno-Care^® on blood lipid parameters. Significant reduction was noted in the overall LDL levels of the treatment group from day 0 to day 28. Perhaps what is more interesting is the increase, though not statistically significant, in HDL levels, compared with a relative decrease in the placebo group.

However, it is the ratios of various lipids and lipid proteins, rather than the absolute values, that are important in assessing cardiovascular risk, and consequently, these ratios were calculated and tabulated.

A significant decrease in the ratio of TC/HDL, and in the ratio of LDL/HDL cholesterol, was noted in the Celt Immuno-Care^® group. A decrease in these ratios corresponds to an associated decrease in the risk of cardiovascular disease (CVD). These ratios are markers for a reduction in the risk of developing atherosclerosis.

Consequently, it is our opinion that these results indicate that Celt Immuno-Care^® could be very beneficial to the health of hypercholesterolemic individuals at risk of developing CVD.

CONCLUSIONS

Celt Naturals Immuno-Care^® and its components appears to have an effect on immune parameters, particularly on basophils and possibly Interleukin-6 levels. Given these changes, Immuno-Care^® would appear to have the potential to substantially alleviate allergic responses.

Celt Immuno-Care^® could also have an effect on autoimmune diseases, such as Crohn’s disease or rheumatoid arthritis, or in the ability of subjects to resist the common cold virus, although studies on these particular populations would be required to verify possible beneficial effects.

This study verified that Celt Naturals Immuno-Care^® supplement is effective in reducing circulating levels of LDL cholesterol and increasing circulating levels of HDL cholesterol.

It is of interest to note that there was a significant decrease in the ratio of TC/HDL and LDL/HDL cholesterol in the Immuno-Care^® group. A decrease in these ratios corresponds to an associated decrease in risk of cardiovascular disease (CVD) as these ratios are markers for a reduction in the risk of developing atherosclerosis. Consequently, these results would be of considerable benefit to the health of hypercholesterolemic individuals at risk of developing CVD.

CELLULAR ANTIOXIDANT PROTECTION STUDY – Sterol 117^™

GITTE S. JENSEN, PHD1 ABSTRACT

The objective of this study was to perform an assessment of the antioxidant protection provided at the cellular level by the product Sterol 117^™ (Celt Naturals, Calgary, Canada). The CAP-e bioassay, using human erythrocytes, was used for this testing, aimed at assessing one important aspect of bioavailability. The water-soluble and water-insoluble components of Sterol 117^™ were tested in parallel. The data showed a dose-dependent protection against intracellular oxidative damage, verifying that antioxidants in Sterol 117^™ were able to protect living human cells from oxidative damage.

Keywords: Antioxidant capacity, bioassay, CAP-e, oxidative damage, plant sterols.

1. INTRODUCTION

The project involved a nutritional product, Sterol 117^™, containing a blend of antioxidants, essential fatty acids, plant sterols, and anti-inflammatory compounds, which is marketed as support of immune function and recovery from fatigue syndromes.

The ingredients include multiple aqueous and non-aqueous sources of antioxidants with various other properties, including immune-activating and anti-inflammatory properties.

Celt Naturals needs to build onto the portfolio of data on the blend of these ingredients. A sequential strategy has been discussed, starting with selected bioassays, and moving towards a human clinical pilot study.

Based on the broad spectrum of potent antioxidants in the blend, the CAP-e antioxidant bioassay was performed, as part of an initial foundation, to gain insight into the antioxidant availability to living cells.

1. Dr. Gitte S. Jensen, PhD (Immunology), Research Director, NIS Labs, 1437 Esplanade, Klamath Falls, Oregon 97601.
2. MATERIALS AND METHODS

2.1. Preparation of Sterol 117^™ for in vitro bioassay work.

Sterol 117^™ (Celt Naturals) was received as an encapsulated dry powder. Immediately prior to testing in biological assays, aqueous and ethanol extracts were prepared from the powder in the following manner:  Five hundred milligrams of powder were added to either 5 mL phosphate-buffered saline (PBS) or 5 mL ethanol. Further dilutions were prepared in PBS.

2.2. Chemicals and reagents.

The following buffers and reagents were obtained from Sigma-Aldrich (St. Louis, MO): phosphate-buffered saline (PBS), RPMI-1640 culture medium, Histopaque 1077, and Histopaque 1119. Dichlorofluorescein diacetate (DCF-DA) was obtained from Molecular Probes (Eugene, OR), a subdivision of Invitrogen (Carlsbad, CA).

2.3. Purification of red blood cells (RBC).

After obtaining informed consent, as approved by the Sky Lakes Medical Center Institutional Review Board (Klamath Falls, OR), peripheral venous blood from healthy volunteers was drawn into sodium heparin and layered onto a double gradient of Histopaque 1119 and 1077. The vials were centrifuged at 2400 rpm for 25 minutes. The RBC fraction was washed twice in PBS, without calcium or magnesium, at 2400 rpm for 10 minutes. The packed RBC was transferred into new vials and again washed twice in PBS, without calcium or magnesium, at 2400 rpm for 10 minutes. The RBC aliquots were stored at 4° Celsius until used in the CAP-e assay.

2.4. Cell-based antioxidant protection of erythrocytes (CAP-e) assays.

The CAP-e assay was conducted, following the method published by Honzel et al (Honzel, Carter, Redman, Schauss, Endres, & Jensen, 2008), modified to an accelerated and more sensitive microplate-based protocol. The RBC cell suspension was prepared for the CAP-e assay by adding 0.1 mL packed RBC into 10 mL PBS. The cell suspension was distributed in a V-bottom 96-well microtiter plate. Twelve wells were not treated with any source of antioxidants and served as negative controls (six wells) and positive controls (six wells) for minimum versus maximum oxidative damage. An additional 12 wells were treated with a standard source of Gallic Acid, a known antioxidant compound, across six different serial dilutions, where each dilution was tested in duplicate. The remaining wells were treated with the isolated compounds from Sterol 117^™, where each compound was tested at six serial dilutions, and each dilution was tested in duplicate.

The RBC were incubated with the potential source of antioxidants for 20 minutes, based on testing of various incubation times on RBC antioxidant uptake of standard antioxidant compounds. Subsequently, antioxidants not absorbed by the cells were removed by washing twice in PBS at 2400 rpm for two-and-a-half minutes. The cells were lysed and the precursor dye was added to the wells. Incubation was performed at room temperature for 15 minutes. Oxidation was carried out in June 2010. Cellular antioxidant protection by Sterol 117^™, using the peroxyl free radical generator AAPH for one hour. A measure of oxidative damage was the intensity of green fluorescence, as measured at 488 nm, using a Tecan (Tecan) SpectraFluor plate reader (40 flashes, optimal gain). The inhibition of oxidative damage was calculated as the reduced fluorescence intensity of product-treated cells, compared to cells treated only with the oxidizing agent. The CAP-e value reflects the IC50 dose of the test product. This was then compared to the IC50 dose of the known antioxidant, Gallic acid. The CAP-e value was expressed as Gallic acid equivalent (GAE) per gram.

3. RESULTS

Cell-based Antioxidant Protection assay Aqueous and ethanol extracts were prepared in parallel, to allow testing of both water-soluble and water-insoluble compounds in the test product.

The graphs in Figure 1 show the results of the CAP-e test on both the PBS and the ethanol extracts of Sterol 117^™. Cells cannot endure more than 2% ethanol, therefore the ethanol extract started at a lower concentration than the PBS extract, and both were serially diluted twofold.

Both extracts showed a dose-dependent antioxidant protection of red blood cells. Thus, we can conclude that both water-soluble and water-insoluble antioxidants are present in the product, in forms that are available to provide protection to live cells. The doses used of the Sterol 117^™ PBS extract allowed the inhibition to reach an IC50. The IC50 is a measure of the effectiveness of a compound in inhibiting (in the case of the CAP-e assay) oxidative damage. If the product is potent enough to show more than 50% inhibition within the dose range tested, then an IC50 can be calculated.

Figure 1. Dose dependent cellular antioxidant protection by Sterol 117^™, when using aqueous
(left panel) versus non-aqueous (right panel) extraction.  The dashed line indicates the level where 50% inhibition was reached (IC50).

The point on the graph where the dashed IC50 line intersects the curve reflects the IC50 dose of the test product, i.e. the dose that provided 50% inhibition of oxidative damage. This IC50 dose is compared to the IC50 dose of the known antioxidant, Gallic Acid (which is used as a control in the assay), resulting in a CAP-e value reported in Gallic Acid equivalent units.

The PBS extract of Sterol 117^™ indeed reached an IC50, giving it a CAP-e value of 1.7 CAP-e units per gram.

Even though the diluted EtOH extract did not reach an IC50 at the doses tested, it was clearly more efficient, i.e. contained more antioxidants per weight, capable of providing a biological protection. This suggests that non-water-soluble antioxidants contribute more to overall antioxidant protection than water-soluble antioxidants. The ethanol extraction is not identical to human digestion, but allows assessment of compounds that are likely to be made available through the digestive process.

The graph below is a comparison of the two extracts from the test product. The ethanol extract shows a higher antioxidant protection, since it provided greater inhibition of oxidative damage to the cells at lower doses than the PBS extract. For example, this can be seen when comparing the antioxidant protection at 1g/L, where the aqueous extract provided 11% protection, in contrast to the 34% protection provided at the same 1 g/L dose of product in an ethanol extract.

Figure 2. Comparison of the cellular antioxidant protection by Sterol 117^™, when using aqueous (solid line) versus non-aqueous (dashed line) extraction.

4. DISCUSSION

Antioxidant capacity of Sterol 117^™ was measured by the recently developed cell-based antioxidant protection (CAP-e) assay [1-4]. Data from the CAP-e assay reflects whether antioxidants can enter into and protect live cells from oxidative damage.

The rationale behind the CAP-e method is important: It allows assessment of antioxidant potential in a method that is comparable to the ORAC test, but only allows measurement of antioxidants that are able to cross the lipid bilayer cell membrane.

As a model cell type, we use the red blood cell (RBC). This is an inert cell type, in contrast to other cell types such as PMN cells, where pro-inflammatory compounds may induce the reactive oxidative burst, or anti-inflammatory compounds may perform cellular signaling and change the behavior of the PMN cell, at doses many times below levels of detection for antioxidants. We developed this assay particularly to be able to assess antioxidants from complex natural products in a cell-based system^[1].

The data showed that Sterol 117^™ contained both water-soluble and water- insoluble antioxidants capable of providing a biologically meaningful antioxidant protection of living cells, in that they were able to protect living human cells form oxidative stress-related damage.  This warrants further clinical study in humans.

REFERENCES

1. Honzel D., Carter S. G., Redman K. A., Schauss A. G., Endress J. R., & Jensen G. S. (2008). Comparison of chemical and cell-based antioxidant methods for evaluation of foods and natural products: generating multifaceted data by parallel testing, using erythrocytes and polymorphonuclear cells. Journal of Agricultural and Food Chemistry, 56(18), 8319-8325.
2. Jensen G. S., Wu X., Patterson K. M., Barnes J., Carter S. G., Scherwitz L., et al. (2008). In vitro and in vivo antioxidant and anti-inflammatory capacities of an antioxidant-rich fruit and berry juice blend. Results of a pilot and randomized, double-blinded, placebo-controlled, crossover study. Journal of Agricultural and Food Chemistry, 56(18), 8326-8333.
3. Schauss A. G., Jensen G. S., Wu X., & Scherwitz L. (2009). Increased antioxidant capacity and inhibition of lipid (delete line break here) peroxidation in healthy adults consuming Monavie Active, an Acai (Euterpe oleracea) fruit-based juice. Acat horticulturae, 841, 97-100.
4. Jensen G. S., Schauss A. G., Beaman R., Ager D. M. (2009). Acai fruit (Euterpe oleracea Mart.): systematic and collaborative study of the phytochemistry, nutrient composition, and in vitro and in vivo bioactivities of the Amazonian palm fruit in humans. Alternative Therapies in Health and Medicine, 15(3), S90-S91.

IMMUNE MODULATION STUDY – Sterol 117^™

Gitte S. Jensen, PhD1 Abstract

Two bioassays, targeted at key aspects of the human innate immune defense, were used to examine the effects of Sterol 117^™ on the function of select subsets of human immune cells. The polar (water-soluble) and non-polar (water-insoluble) components of Sterol 117^™ were tested in parallel. The data showed that Sterol 117^™ activated human natural killer cells in vitro and contains compounds that support phagocytosis, as an important part of antibacterial immune defense. Keywords: Immune modulation, Natural Killer cells, Phagocytes, antiviral, antibacterial.

1. INTRODUCTION

The project involved a nutritional product, Sterol 117^™, containing a blend of antioxidants, essential fatty acids, plant sterols, and other anti-inflammatory compounds, and which is marketed as support of immune function and recovery from fatigue syndromes. The ingredients include multiple aqueous and non-aqueous sources of antioxidants, with various other properties, including immune-activating and anti-inflammatory properties.

The initial testing, conducted at NIS Labs, showed that the product contains antioxidants capable of protecting live cells from free radical damage when the cells were exposed to oxidative stress. An additional pilot test provided data that suggested support of antiviral immune defense mechanisms, since both the aqueous and ethanol extracts induced activation of Natural Killer (NK) cells.

This report presents data that further documents NK cell activation by Sterol 117^™ in more detail, regarding this important aspect of antiviral immune defense. The report also documents the effect on a key aspect of our antibacterial immune defense mechanisms; phagocytosis.

1. Dr. Gitte S. Jensen, PhD (Immunology), Research Director, NIS Labs, 1437 Esplanade, Klamath Falls, Oregon 97601.
2. MATERIALS AND METHODS

2.1. Preparation of Sterol 117^™ for in vitro bioassay work.

Sterol 117^™ (Celt Naturals, Calgary, Canada) was received as an encapsulated dry powder. Immediately prior to testing in biological assays, aqueous and ethanol extracts were prepared from the powder in the following manner: Five hundred milligrams powder was added to either 5 mL phosphate-buffered saline (PBS) or 5 mL ethanol. Further dilutions were prepared in PBS at physiological pH.

2.2. Chemicals and reagents.

The following buffers and reagents were obtained from Sigma-Aldrich (St. Louis, MO): phosphate-buffered saline (PBS), RPMI-1640 culture medium, Histopaque 1077, and Histopaque 1119. Dichlorofluorescein diacetate (DCF-DA) was obtained from Molecular Probes (Eugene, OR), a subdivision of Invitrogen (Carlsbad, CA).

2.3. Purification of white blood cells.

After obtaining informed consent, as approved by the Sky Lakes Medical Center Institutional Review Board (Klamath Falls, OR), peripheral venous blood from healthy volunteers was drawn into sodium heparin and layered onto a double-gradient of Histopaque 1119 and 1077. The vials were centrifuged at 2400 rpm for 25 minutes. The peripheral blood mononuclear cell (PBMC) and polymorphonuclear (PMN) fractions were harvested and washed twice in PBS, without calcium or magnesium, at 2400 rpm for 10 minutes. The cells were used immediately for testing in the NK activation and phagocytosis bioassays.

2.4. Evaluation of activation status of human NK cells in vitro.

Freshly purified human PBMC were used for these assays. The cells were plated in micro-well plates. Negative control samples in quadruplicate were left untreated. Positive control samples were treated with Interleukin-2 (IL-2). Two sets of samples were treated with serial dilutions of test product. One set was only treated with test product, to evaluate the direct effect of the test product on NK cell activation. The second set was also treated with IL-2, to see whether the test product affects the response of NK cells to the known NK cell activator IL-2.

After 18 hours of culture, cells were stained with fluorescent markers for the T cell antigen CD3, the antigen CD56, and the activation molecule CD69. The expression level of CD69 on the surface of CD3-negative and CD56-positive NK cells was analyzed, as a measure of mean fluorescence intensity (MFI). The analysis allows us to detect if compounds in a test product directly activate NK cells in vitro. It also provides indication of whether a test product enhances or otherwise affects the response of NK cells to IL-2. This would be an important indication of whether the product may support an already ongoing antiviral immune response.

2.5. Evaluation of phagocytic activity.

Freshly purified peripheral blood PMN cells were pretreated with test products for three minutes, and then introduced to fluorescent micro-particles, mimicking August 2010. Immune modulation by Sterol 117^™ bacteria. The cells were allowed to ingest particles for two minutes, after which free micro-particles were removed by centrifugation. The fluorescence intensity of phagocyte cells was then evaluated by flow cytometry.

3. RESULTS

Effect on activation of NK cells: CD69 expression levels. Treatment of PBMC, which is the white blood cell fraction containing the NK cell subset, resulted in increased expression of CD69 on the surface of the NK cells. Both S117 PBS and S117 EtOH increased the expression of CD69 on the cell surface of CD3- CD56+ NK cells.

This effect was dose-dependent for both product preparations, with S117 EtOH performing better than S117 PBS. For S117 EtOH, the three highest concentrations of product, and for S117 PBS the two highest concentrations.

The treatment with test product may result in more phagocytes deciding to engage in phagocytosis, and may also lead to a faster or stronger rate of phagocytosis, resulting in higher numbers of fluorescent micro-particles per cell. The data analysis examined both aspects of the phagocytic activity of the cells produced statistically significant increases in CD69 expression above baseline (P<0.05). When the effects of S117 PBS and S117 EtOH on NK cell activation were assayed in the presence of the known NK cell activator IL-2, both products showed synergy with IL-2 and increased CD69 expression to levels higher than those resulting from IL-2 treatment alone. This effect was very pronounced with the S117 EtOH product, where the 0.25 g/L and 0.063 g/L doses resulted in a doubling of CD69 expression on NK cells (P<0.02).

Figure 1. A) Sterol 117^™ directly activates human NK cells in vitro. Dose-dependent activation of NK cells by Sterol 117^™ was seen both when using aqueous (PBS) versus non-aqueous (EtOH) extraction. B) Synergistic enhancement of NK cell activation in the presence of IL-2. The dotted line indicates the baseline level of CD69 expression on the NK cells. The broadly dashed horizontal line indicates the level of CD69 expression achieved when NK cells were activated by the known stimulus IL-2. In addition to evaluating the mean fluorescence intensity (MFI) of the CD69 activation marker, analysis was also performed to evaluate if the relative number of
CD69-positive NK cells changed with treatment by test product. The highest dose of S117 PBS also produced a statistically significant increase in the percent of NK cells result when NK cells were treated with expressing CD69 that was greater than IL-2 alone (P<0.05). 

Figure 2. Sterol 117^™ increases the numbers of NK cells that express the CD69 marker, both in a direct manner (top graph) and in synergy with IL- 2 (bottom graph). A dose-dependent increase was seen both when using aqueous (PBS) versus non-aqueous (EtOH) extraction.

Effect on activation of Natural Killer T (NKT) cells

Cell subset, called NKT cells, is almost in most normal healthy human donors, non-detectable. During analysis, we the proportion of a rare CD3+ CD56+ found that two out of the three donors had detectable levels of NKT cells. This allowed us to gather pilot data on how the test product may affect the activation status of this type of cell, known to conduct rapid immune-modulating effects, including cytokine secretion.

Expression of CD69 on NKT cells was increased by treatment of cells with S117 PBS and S117 EtOH.

In the presence of IL-2, S117 EtOH nearly quadrupled the expression level of CD69 on NKT cells when assayed at the 0.25 g/L concentration. This increase was much higher than the effect of IL-2 alone, and was highly statistically significant (P<0.003).

Figure 3. A) Sterol 117^™ directly activates human NKT cells in vitro. Dose-dependent activation of NKT cells by Sterol 117^™ was seen both when using aqueous (PBS) versus ethanol (EtOH) extraction. B) Synergistic enhancement of NTK cell activation in the presence of IL-2 was seen for the EtOH extract.

Phagocytic activity.

When measuring the percent of PMN cells that were actively engaged in phagocytic activity, it was seen that treatment with S117 EtOH, at the two highest doses tested, showed an effect on increasing this percent. Treatment of cells with S117 EtOH also increased the number of particles ingested per phagocytic PMN cell (indicated by an increase in fluorescence intensity).

Figure 4. The treatment of human phagocytes with the EtOH extract of Sterol 117^™ resulted in an enhanced function of these cells, in terms of the antibacterial behavior of engulfing foreign particles.

4. DISCUSSION

Our body’s primary defense mechanisms towards cancers and viral diseases involve a group of cells called NK cells. These cells travel in our bloodstream in a state of rest, but can be immediately recruited into tissues by chemical signals, and activated through various mechanisms to a) kill cancer cells, b) divide and make more NK cells, and c) secrete substances that attract other cells into the site.

The data showed that Sterol 117^™ contained both water-soluble and water-insoluble antioxidants, capable of providing a significant increase in the activation marker CD69 on both NK cells and NKT cells. Expression of the CD69 marker on NK cells has been equated to an increased cytotoxic capacity, in terms of the capacity of the NK cells to kill transformed target cells^[1].

Phagocytosis of microbial particles is an important part of the innate (immediate) immune response. It is a rapid process, and the effect of test products on enhancing this cellular
function is often almost immediate. Sterol 117^™ contains compounds that are extractable by ethanol, i.e. non-polar compounds, that induce an enhanced phagocytic capacity in vitro.  This may suggest that ingestion of Sterol 117^™ supports phagocytes in the gut mucosal lining, and could possibly be supportive of antibacterial defense mechanisms at the gut interface.

These findings, taken together, suggest that Sterol 117^™ contains compounds that are capable of supporting two distinct functions of the human innate immune defense.

REFERENCES

1. Clausen J, Vergeiner B, Enk M, Petzer AL, Gastl G, Gunsilius E. Functional significance of the activation-associated receptors CD25 and CD69 on human NK-cells and NK-like T-cells. Immunobiology. 2003;207(2):85-93.