CLINICAL AND EXPERIMENTAL STUDIES
Natural Killer (NK) Cell Activity
Peripheral blood mononuclear cells were isolated and pooled from several healthy donors. Sixty thousand cells were added to each well of 96-well microtiter plate. Various immune-modulating ingredients, including 4Life Transfer Factor Tri-Factor Formula, were added to select wells on the plate and the 48-hour incubation started. At end of the incubation period, 30,000 K562 cells were added to each well. MTT assay techniques were used to determine the cytotoxic index. The various 4Life Transfer Factor products resulted in cytotoxic indices of 80-98%. By comparison, mononuclear cells incubated with IL-2 for the same 48-hour period produced a cytotoxic index of 88%.(18)
The NK cell activity of 4Life Transfer Factor has more recently been evaluated via flow cytometry technique. PBMCs derived from six healthy donors were plated in 96-well plates at 96,000/90 μl concentration. UltraFactor XF was solubilized in PBS to 10 mg/ml and added to PBMCs at final concentrations of 10, 100, and 1000 μg/ml. K562 cells were added into wells at 4000 per 90 μl and incubated for 48 hours. IL-2 at 20 ng/ml was used as positive control. Results revealed that UltraFactor XF's cytotoxicity against K562 cells was greater than IL-2 in four out of the six donors and similar to IL-2 in one out of the six donors.(19)
CD4 T-Helper Cell Research
Multiple studies were performed using an FDA-approved diagnostic CD4 T-Helper cell assay kit and/or a T-Cell Memory (CD8) assay kit under development by the same company. Similar to the NK cell research described above, these ex vivo studies were performed on 96-well microtiter plates measuring ATP production via a luciferase-based luminescence reaction. The CD4 assay utilized Phytohaemagglutinin (PHA)-stimulated cells isolated from whole blood via the use of Dynabeads™. An 18-hour incubation of these isolated, stimulated CD4 cells with 4Life Transfer Factor products resulted in a modulation of immune cell activity as exhibited by a decrease in Adenosine Triphosphate (ATP) production without a negative impact on cell viability. It is hypothesized that this reduction on ATP production is a result of a redirection in immune cell focus, essentially diminishing the distraction induced by the addition of PHA to the microtiter wells.(20)
Salivary Secretory IgA (SIgA)-Preliminary Investigation
Twenty-four subjects naïve to 4Life Transfer Factor supplementation were enrolled in a small-scale, preliminary test. Twenty-one were included in the final analysis. Salivary samples were collected from each subject weekly at roughly the same time of day and day of the week. Saliva was collected over a five-minute period via passive drool while subjects chewed on a piece of Parafilm™. The samples were put on ice and then frozen at -70°C until assay. The commercial Salimetrics™ salivary IgA assay kit was used for analysis. Subjects were given 4Life Transfer Factor Tri-Factor Formula at 2 capsules per day for two weeks and then transitioned to 4Life Transfer Factor RioVida Tri-Factor Formula at 60 ml per day for an additional two weeks. At the end of the four-week supplementation period, the group showed an average 73% increase in salivary secretory IgA (SIgA) production over their baseline value. Furthermore, none of the 21 subjects showed a SIgA production rate less than their baseline value at the end of the test.(21)
A study conducted with 30 college students found that either 1× 15 days or 2× 15 days (with a two-week break in between) of 4Life Transfer Factor Classic administered according to label dose helped them maintain their health. In both groups, product administration improved the number of CD8+ T-cells and NK cells to healthier levels. Particularly, those who took the product for 2×15 days showed prolonged health maintenance and improvement of immune cell markers than those who took it for 15 days. Specifically, the maintenance of good health and improvement of immune cell markers remained for up to three months after stopping product administration in those who took the product for 2× 15 days, in comparison to one month in those who took the product for 1×15 days. (22)
In a study of acute toxicity, rats were assessed for 14 days following a single gavage of 4Life Transfer Factor Tri-Factor Formula. Five female SD rats were each gavaged with a dose of 2,000 mg/kg. No treatment-related mortalities occurred and there were no clinical signs of toxicity. No significant difference in body weight occurred. No gross lesions were found at necropsy in any of the animals. Thus, acute toxicity is considered to be greater than 2,000 mg/kg (human equivalent dose of approximately 320 mg/kg). (23)
Another similar single-dose oral toxicity study was conducted in mice. Six female Wistar mice each received 2,000 mg/kg of 4Life Transfer Factor Chewable Tri-Factor Formula via oral gavage and monitored for 14 days. No observable toxicity occurred, as assessed by mortality, body weight gain, histopathology of brain, liver, kidneys, and lungs, or clinical signs of aggression, lethargy, breathing difficulties, diarrhea, mobility, or shivering. Thus, the no-observed adverse effect level was considered to be greater than 2,000 mg/kg in mice, which is equivalent to approximately 9.7 g/day in humans. (24)
Recent toxicity studies performed by an independent toxicology laboratory were conducted to evaluate the mutagenicity and genotoxicity potential of UltraFactor XF (colostrum ultrafiltrate). Mutagenicity was assessed by the Bacterial Reverse Mutation assay. Results revealed that the test article has no mutagenic activity at any of the concentrations tested. Genotoxicity was assessed by the Mammalian Chromosome Aberration test. Results demonstrated that the test article, tested up to the maximum recommended concentration, did not induce structural chromosome aberrations in this mammalian system. The laboratory concluded that UltraFactor XF is considered not clastogenic in this system. (25)
Oral toxicity studies performed by the same toxicology laboratory assessed the potential short-term and long-term toxicity of UltraFactor XF in rats. In both, 14-day and 90-day repeated dose studies, male and female Wistar rats received by oral gavage 1050, 2100, or 4200 mg/kg body weight/day of UltraFactor XF or placebo. Results revealed that no mortality occurred at any given dose. Clinical observations showed no adverse effect of the test article on behavior and physical condition of the animals. No abnormal body weight gain or food consumption was observed. Ophthalmological and hematological evaluations demonstrated no adverse effect by the test article. Similarly, no changes were observed in clinical chemistry, gross pathology, organ weight, or histopathology at any given dose. It was concluded that the no-observed adverse effect level was greater than 4200 mg/kg in rats. This dose is equivalent to 40 g/day in humans (25).
An expert panel of toxicologists have evaluated the above-mentioned toxicity data and concluded that UltraFactor XF is generally recognized as safe (GRAS). (26)
The use of transfer factors is contraindicated in people receiving immunosuppressive therapy, though actual interactions have not been documented. The use of transfer factors during pregnancy and nursing has not been evaluated.
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