Compositions including different types of transfer factor

Abstract

A composition for eliciting a T-cell mediated immune response in a subject includes transfer factor from at least two different types of source animals. For example, the composition may include mammalian transfer factor and nonmammalian transfer factor. An example of the composition includes a combination of a colostrum-derived product, which includes the mammalian transfer factor, and an egg-derived product, which includes the nonmammalian transfer factor. Additionally, the egg-derived product may be substantially free of fat. Methods for forming the composition and eliciting T-cell mediated immune responses in subjects that have been treated with the composition are also disclosed.

Claims

1. A component consisting of: a fraction and/or an extract of bovine colostrum comprising mammalian transfer factor; and chicken egg yolk comprising avian transfer factor.

2. The component of claim 1, wherein the fraction and/or extract of bovine colostrum is a fraction of bovine colostrum.

3. The component of claim 2, wherein the fraction of bovine colostrum is substantially free of fat.

4. The component of claim 3, wherein the fraction of bovine colostrum is substantially free of at least one of casein, cells, cell debris, antibodies, and allergenic agents.

5. The component of claim 1, wherein the fraction and/or extract of bovine colostrum is an extract of bovine colostrum.

6. The component of claim 1, wherein the chicken egg yolk is a fraction of chicken egg yolk.

7. The component of claim 1, wherein the chicken egg yolk is an extract of chicken egg yolk.

8. The component of claim 1, wherein the fraction and/or extract of bovine colostrum is dried bovine colostrum and the chicken egg yolk is dried chicken egg yolk.

9. The component of claim 1, wherein the fraction and/or extract of bovine colostrum and the chicken egg yolk of the component are selected, and amounts and relative proportions of the fraction and/or extract of bovine colostrum and the chicken egg yolk in the component are tailored, to synergistically elicit an elevated cell-mediated immune response in a treated subject.

10. The component of claim 9, wherein the relative proportions of the fraction and/or extract of bovine colostrum and the chicken egg yolk in the component are further tailored to maintain the elevated cell-mediated immune response for a period of at least 48 hours.

11. A component consisting of: a fraction and/or an extract of bovine colostrum including mammalian transfer factor; and a fraction and/or extract of chicken egg yolk including avian transfer factor.

12. The component of claim 11, wherein the fraction and/or extract of bovine colostrum and the fraction and/or extract of chicken egg yolk are in dry form.

13. The component of claim 11, wherein amounts and relative proportions of the fraction and/or extract of bovine colostrum and the fraction and/or extract of chicken egg yolk are tailored to synergistically elicit an elevated cell-mediated immune response in a treated subject.

14. The component of claim 13, wherein the relative proportions of the fraction and/or extract of bovine colostrum and the fraction and/or extract of chicken egg yolk are further tailored to maintain the elevated cell-mediated immune response for a period of at least 48 hours.

15. A component consisting of: a fraction and/or extract of a source of mammalian transfer factor, said fraction and/or extract comprising mammalian transfer factor; and an avian transfer factor-containing component.

16. The component of claim 15, wherein the fraction and/or extract of the source of mammalian transfer factor and the avian transfer factor-containing component are in dry form.

17. The component of claim 15, wherein amounts and relative proportions of the fraction and/or extract of the source of mammalian transfer factor and the avian transfer factor-containing component are selected to synergistically elicit an elevated cell-mediated immune response in a treated subject.

18. The component of claim 17, wherein the relative proportions of the fraction and/or extract of the source of mammalian transfer factor and the avian transfer factor-containing component are further tailored to maintain the elevated cell-mediated immune response for a period of at least 48 hours.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) In the drawings, which depict exemplary embodiments of various aspects of the present invention:

(2) FIG. 1 depicts an example of the manner in which a composition that incorporates teachings of the present invention may be embodied;

(3) FIG. 2 is a schematic representation of capsulation equipment that may be used to introduce a powdered embodiment of the composition of the present invention into gelatin capsules; and

(4) FIG. 3 schematically illustrates an exemplary test protocol that was conducted to determine the efficacy of various aspects of the present invention.

DETAILED DESCRIPTION

(5) An exemplary embodiment of composition that incorporates teachings of the present invention includes transfer factor from at least two different types of source animals. By way of nonlimiting example, a composition according to the present invention may include mammalian transfer factor and nonmammalian transfer factor.

(6) The different types of transfer factor of the inventive composition may be obtained from any suitable source. For example, mammalian transfer factor may be obtained from colostrum, as described in Wilson or otherwise as known in the art (e.g., a leukocyte (white blood cell) extract, a splenic (i.e., “from the spleen”) extract, etc.). An exemplary source for nonmammalian transfer factor is an egg of an animal, such as a chicken, as described in Hennen. Thus, a composition according to the present invention may include a first component which comprises colostrum or a fraction or extract thereof, which are collectively referred to herein as a “colostrum-derived product,” as well as a second component that comprises egg or a fraction or extract thereof, which are also referred to herein as an “egg-derived products.”

(7) As compositions that incorporate teachings of the present invention include transfer factor from different types of source animals, they may include transfer molecules with a broader array of antigen-specificity or pathogen-specificity than conventional transfer factor-containing compositions. Thus, a composition according to the present invention is capable of enlisting the immune system of a treated animal to elicit a T-cell mediated immune response against a broader array of pathogens than those against which conventional transfer factor-containing compositions are effective. This is because different types of animals may be exposed to different types of antigens or pathogens, such as by vaccination, the animals' environments, or the like. Moreover, it is known that some conditions in certain animals are caused by multiple infections, even further expanding the specificity of a composition according to the present invention. For example, one or more pathogens may adversely affect (e.g., suppress or monopolize) the host's immune system, while one or more other pathogens may be permitted to cause a disease state in the host. As another example, some disease states are caused by a combination of pathogens.

(8) As an example, a composition which includes transfer factor-containing components from both cows and chickens will include transfer factor molecules which are specific to antigens or pathogens to which cows are exposed, as well as transfer factor molecules that have specificity for antigens or pathogens to which chickens are exposed. As both cows and chickens may be exposed to antigens or pathogens to which the other is not exposed, such a composition may include transfer factor molecules with antigen or pathogen specificities that would not be present in a composition that includes only transfer factor from cows (e.g., by way of a colostrum-derived product) or transfer factor from chickens (e.g., through an egg-derived product).

(9) A composition of the present invention may include about the same amounts, measured in terms of weight or volume, of a colostrum-derived product and an egg-derived product (i.e., about 50% colostrum-derived product and about 50% egg-derived product). Alternatively, a composition that incorporates teachings of the present invention may include more colostrum-derived product (e.g., about 85% or 60%, by combined weight of the colostrum-derived product and egg-derived product) than egg-derived product (about 15% or 40%, by weight). As another alternative, the inventive composition may include more egg-derived product (e.g., about 60% or 85%, by weight) than colostrum-derived product (e.g., about 40% or 15% by weight). As another example, a composition that incorporates teachings of the present invention may include about one percent, by weight, of one of a colostrum-derived product and an egg-derived product and about 99%, by weight, of the other of the colostrum-derived product and the egg-derived product. Although specific amounts of colostrum-derived product and egg-derived product have been provided, any combination thereof is within the scope of the present invention.

(10) In addition to including a source of transfer factor (e.g., a colostrum-derived product, an egg-derived product, etc.) a composition that incorporates teachings of the present invention may include one or more other ingredients, including, but not limited to, vitamins, minerals, proteins, natural products (e.g., herbs, mushrooms, roots, etc., or extracts thereof), and the like. Additional ingredients may be useful for providing further advantages to subjects to which the composition is administered, or may enhance the ability of the transfer factor in the composition to elicit or enhance a secondary, or delayed-type hypersensitivity, immune response.

(11) As shown in FIG. 1, without limiting the scope of the present invention, a composition 10 according to the present invention may take the form of a powdered or particulate substance, which includes the multiple types of transfer factor (not shown). In order to ensure that an appropriate and precise dosage of composition 10 is administered to a subject (not shown), composition 10 may be contained within a gelatin capsule 12 of a type which is well-known and readily available to those in the art. The result is the illustrated capsule 14. Alternatively, a composition according to the present invention may be embodied as tablet, a so-called “caplet,” an unencapsulated powder, a liquid, a gel, or in any other pharmaceutically acceptable form. Suitable processes for placing the inventive composition into any such form are readily apparent to those of skill in the art.

(12) In an exemplary embodiment of a method for making or forming a composition according to the present invention, a first type of transfer factor may be combined with a second type of transfer factor. Additionally, one or more other types of transfer factor may be combined with the first and second types of transfer factor. The different types of transfer factor that are combined may be substantially purified transfer factor, components or “products” that include transfer factor, or any combination thereof.

(13) Turning again to FIG. 2, a process for forming composition-filled capsules 14, such as that shown in FIG. 1, is provided merely as an example for a method for making a composition that incorporates teachings of the present invention. As illustrated, the composition 10 is made and composition-filled capsules 14 are formed using standard capsulation equipment 20 of a type known in the industry, such as the SF-135 capsule filling machine available from CapPlus Technologies of Phoenix, Ariz.

(14) In addition to one or more composition supply hoppers 24, an auger 26 associated with each composition supply hopper 24, and a feed station 28 with which each auger 26 and the conduit 27 within which auger 26 is contained communicates, capsulation equipment 20 includes one or more capsule hoppers 30, as well as a pneumatic feed system 32 for transporting capsule bodies 12a and/or caps 12b to feed station 28.

(15) As the capsulation equipment will introduce the mixture into capsules, which may be swallowed by a subject, it is currently preferred that the substantially fat-free component and the egg-derived product be introduced into the capsulation equipment in powdered form. The substantially fat-free component dilutes the amount, or concentration, of fat (e.g., from egg yolk) present in the mixture relative to the concentration of fat which is present in the egg-derived product. Accordingly, the relative amounts of substantially-fat free product and the egg-derived product may be tailored to provide a fat concentration that will minimize clogging of the capsulation equipment.

(16) Continuing with the example of a composition 10 which includes a colostrum-derived product 10a as the substantially fat-free component and an egg-derived product 10b, colostrum-derived product 10a and egg-derived product 10b may be introduced simultaneously into a single composition supply hopper 24 of capsulation equipment 20. For example, colostrum-derived product 10a and egg-derived product 10b may be mixed upon introduction thereof into composition supply hopper 24, as shown, or premixed. By introducing a substance which has a lower fat content than egg-derived product 10b into composition supply hopper 24 along with egg-derived product 10b, the fat content (e.g., concentration) of the resulting mixture is less than that of egg-derived product 10b, reducing or eliminating the likelihood that composition supply hopper 24, auger 26, conduit 27, feed station 28, or any other component of capsulation equipment 20 will be coated with cholesterol or fat.

(17) Following introduction of a predetermined amount of composition 10 into capsule bodies 12a at feed station 28, the filled capsule bodies 12a are transported to a capsule closing station 34, where capsule caps 12b are assembled therewith to fully contain composition 10 within capsule 12.

(18) Again, a composition-filled capsule 14 is only one example of the manner in which a composition that incorporates teachings of the present invention may be embodied. The inventive composition may also take other forms, such as tablets, caplets, loose powder, liquid, gel, liquid-filled or gel-filled capsules, any other pharmaceutically acceptable form known in the art, each of which may be made by known processes.

(19) The composition of the present invention may be administered to a subject (e.g., a mammal, such as a human, a dog, or a cat, a bird, a reptile, a fish, etc.) by any suitable process (e.g., enterally, parenterally, etc.), depending, of course, upon the form thereof. For example, virtually any form of the composition (e.g., a capsule, tablet, caplet, powder, liquid, gel, etc.) may be administered orally (i.e., through the mouth of the subject), provided that the composition includes a pharmaceutically acceptable carrier of a type known in the art that will prevent degradation or destruction of transfer factor molecules by the conditions that persist in the digestive tract of the subject without substantially interfering with the efficacy of the transfer factor molecules included in the composition.

(20) The dosage of composition or transfer factor within the composition that is administered to the subject may depend on a variety of factors, including, without limitation, the subject's weight, the health of the subject, or conditions (e.g., pathogens) to which the subject has been exposed.

(21) Administration of the composition to the subject may cause the immune system of the subject to elicit a T-cell mediated immune response against one or more antigens or pathogens. Thus, the composition may be administered to a subject to treat a disease state that the subject is experiencing, to prevent the subject from exhibiting a disease state caused by a particular pathogen, or to merely enhance the overall health of the subject's immune system and abilities to fight off infecting or invading pathogens.

(22) The following EXAMPLES illustrate the enhanced ability of a composition which includes transfer factors from multiple types of source animals to cause an immune system of a treated subject to elicit a T-cell mediated immune response to various types of pathogens, in the form of target cells. The ratios used in the EXAMPLES are based on the weight of the material (e.g., egg powder, colostrum powder) used in a particular test sample.

Example 1

(23) In EXAMPLE 1, a preliminary test, the target cells included bacteria (e.g., C. pneumoniae and H. pylori) and viruses (e.g., herpes simplex virus-1 (HSV-1) and herpes simplex virus-2 (HSV-2)) in the form of virally infected cells, as well as to cancerous, or malignant, cells (e.g., K562 erythroleukemic cells).

(24) The in vitro technique that was used to make these determinations was the so-called “chromium-51 release assay,” which includes measurement of the amount of radioactive chromium-51 (Cr-51) released by cells that have been attacked by NK cells. The radioactivity measurement may be obtained, for example, with a Beckman 2000 Gamma Counter, which is available from Beckman Coulter, Inc., of Fullerton, Calif.

(25) In EXAMPLE 1, which was a preliminary test, a fixed amount (5 micrograms per milliliter of nutrient media and cellular milieu) of a powdered composition was provided in the nutrient media and cellular milieu, along with a substantially fixed amount of NK cells. Examples of the powdered compositions that were used include bleached wheat flour, Transfer Factor™ (TF), available from 4Life Research, LLC, of Sandy, Utah, Transfer Factor Plus' (TFP or TF+), also available from 4Life Research, avian transfer factor available in a lyophilized (i.e., freeze-dried) whole egg powder, and mixtures of TF and TFP (both the formula marketed in the United States and that marketed internationally) with avian transfer factor in a ratio of about 85% TF or TFP (i.e., bovine transfer factor), by weight, to about 15% avian transfer factor, by weight. The powdered composition, nutrient media, NK cells, and target cells were mixed and incubated for four hours prior to measuring the radioactive atoms that were released by disruption of the target cells by the NK cells. Each exemplary reaction was conducted in triplicate, with the results of the three reactions having been averaged.

(26) In addition to including one or more types of transfer factor, TFP includes a variety of other components, including maitake and shiitake mushrooms, cordyceps, inositol hexaphosphate, beta glucans, beta sitosterol, and olive leaf extract. Maitake and shiitake mushrooms are known to be good sources for polysaccharides and to promote T-cell function. Cordyceps are also rich in polysaccharides. Beta glucans, another class of polysaccharides, is also known to be an important immune cell stimulator.

(27) The following TABLE includes data of the counts per minute obtained with each combination of target cells and powdered composition, as well as the effectiveness of each powdered composition in eliciting an NK cell-mediated immune response against the target cells relative to the NK cell-mediated immune response relative to (measured in percent increase) the same types and concentrations of target cells in the presence of bleached wheat flour.

Example 1

(28) TABLE-US-00001 TABLE 1 Target Cells C. Pneu H. Pyl K562 HSV-1 HSV-2 Spontaneous 1,256/ 1,875/ 1,620/ 974/ 1,476/ Flour 1,323/ 1,121/ 1,267/ 2,017/ 1,262/ Average Composition 1,290/ 1,498/ 1,444/ 1,496/ 1,365/ TF 2,593/ 2,499/ 2,445/ 2,240/ 2,473/ % increase over flour  96% 123%  93%  11%  96% % increase over average 101%  67%  69%  50%  81% TFP 3,386/ 2,701/ 3,243/ 2,944/ 1,956/ % increase over flour 156% 141% 156%  46%  55% % increase over average 163%  80% 125%  97%  43% Bov-Av TF 14,857/  11,434/  6,639/ 17,910/  10,626/  % increase over flour 1023%  920% 424% 788% 742% % increase over average 1052%  663% 360% 1098%  679% Bov-Av TFP 6,196/ 5,543/ 4,008/ 8,050/ 4,693/ US % increase over flour 458% 485% 306% 389% 362% % increase over average 380% 270% 178% 438% 244% Bov-Av TFP 5,747/ 4,786/ 3,640/ 7,366/ 4,269/ Intl % increase over flour 424% 417% 277% 355% 328% % increase over average 346% 219% 152% 393% 213% 100% Avian 2,553/ 1,860/ 2,483/ 2,985/ 2,183/ TF % increase over flour  93%  66%  96%  48%  73% % increase over average  98%  24%  72% 100%  60%

(29) Notably, the formulations denoted “TFP” include only about half (0.466667) of the transfer factor as that present in the formulations denoted “TF.” Accordingly, one of ordinary skill in the art would expect the data that corresponds to cytotoxicity induced by the products identified as “Bov-Av TFP US” and “Bov-Av TFP Intl” to be somewhat less than the cytotoxicity induced by the product identified as Bov-Av TF. Instead, these numbers were much higher. In fact, it appears that the data that corresponds to “Bov-Av TFP US” and “Bov-Av TFP Intl” is about ten times too high. Accordingly, appropriate corrections have been made to TABLE 1. Additionally, further testing has been conducted, as is evident from the ensuing EXAMPLES, to evaluate and verify the abilities of combinations of different types of transfer factor to elicit T-cell responses in treated animals.

(30) The preliminary results that are set forth in TABLE 1 show that administration of a composition of the present invention to a subject will likely increase the subject's secondary, or delayed-type hypersensitivity, immune response, as effected by NK cells, against one or more pathogens to a degree which far exceeds the NK cell activity initiated by both colostrum-derived transfer factor and egg-derived transfer factor alone. In fact, the results show that a composition that incorporates teachings of the present invention may result in facilitation of the activity of NK cells with an unexpected degree of synergy.

(31) In view of these results, further experimentation was conducted to determine the efficacy of a broader range of aspects of the present invention.

Example 2

(32) The effects of various transfer factor compositions, including compositions that incorporate teachings of the present invention, on the activity of lymphocytes in attacking cancer cells was evaluated. FIG. 3 schematically represents the protocol for the evaluation. Blood from healthy donors was obtained, at reference 40. Mononuclear cells, including natural killer cells, were separated from other constituents of the blood, at reference character 42, by standard phycol-urographin methodology, employing a density gradient p=i, 077 g/cm.sup.3. The isolated mononuclear cells, or “effector cells,” at a dilution of about 60,000 cells/100 μl of culture medium, were then introduced in 100 μl aliquots into the wells of a 96-well microtitre plate, such as that available from Corning Incorporated of Corning, N.Y., under the trade name COSTAR®, as shown at reference character 44.

(33) Thereafter, transfer factor-containing test samples, or “additives,” as noted in TABLES 2 through 5 below, were introduced into each well, with resulting concentrations of transfer factor in the test samples being 1 mg/ml, 0.1 mg/ml, 0.01 mg/ml, 0.001 mg/ml, 0.0001 mg/ml, and 0.00001 mg/ml, as is also shown at reference character 44. A control including no transfer factor product was also employed. The microtitre plates were then placed in a CO.sub.2-incubator with conditions of 5% CO.sub.2 atmosphere, 100% humidity, and a temperature of 37° C., and incubated for periods of 24 hours and 48 hours. Each study variant was conducted in triplicate.

(34) After incubation, about 30,000 K-562 tumor cells (i.e., erythroblastotic human leukemia), or “target cells,” were introduced into each well, as illustrated at reference character 46, providing a ratio of effector cells-to-target cells of about 2:1. The effector and target cells were then incubated for periods of 18 hours and 24 hours in the CO.sub.2 incubator, under the same conditions listed above.

(35) Thereafter, at reference character 48, the MTT method of defining the viability of cellular cultures, which employs a soluble yellow bromide, 3-(4,5-dimethylthiasol-2-il)-2,5-tetrazol (MTT), was used to determine the number of K-562 tumor cells that were killed in each well. In such a test, live cells reduce the MTT to insoluble purple-blue intracellular crystals of MTT-formazan (MTT-f). Nonviable dead cells are not capable of reducing the MTT to MTT-f. Thus, the optical properties of the resulting solution may be evaluated to provide an indication of the affect of various transfer factor-containing products on the ability of the effector cells to kill the K-562 tumor cells. More specifically, the intensity of MTT transformation into MTT-f reflects the general level of the studied cells' dehydrogenase activity and is modulated by the activity of conjugated fermentation systems; e.g., respiratory chain of electrons transmission, etc.

(36) The MTT solution used in this EXAMPLE was prepared in 5 mg/ml of Henks' saline solution, as known in the art. Equal volume aliquots of the MTT solution were introduced into the wells of the microtitre plates, and the plates were incubated in a CO.sub.2 incubator, under the same conditions noted above, for a period of about three to about four hours. The microtitre plates were then centrifuged at about 1,500 rpm for about 5 minutes, the supernatant was removed, and 150 μl aliquots of dimethylsulfoxide (DMSO) were introduced into the wells.

(37) The microtitre plates were then permitted to sit at room temperature for a period of thirty minutes, allowing formazan crystals to completely dissolve. Thereafter, a multiwell spectrophotometer (LABSYSTEMS MultiScan MSS 340, available from Cambridge Scientific Products of Cambridge, Mass.) was used to evaluate each well of each microtitre plate at a wavelength of 540 nm.

(38) As shown at reference character 50, the optical density (OD) measurements that were obtained with the spectrophotometer were then used to calculate the cytotoxic index (%) (CI (%)) of each well. The CI (%) calculation was performed according to the standard formula:
Cl(%)=[I−(O.sub.e+t−OD.sub.e)/OD.sub.t]*100,
where ODe+t is the OD in experimental series, ODe is the OD in wells including only effector cells, and OD.sub.t is the OD in the wells including only target cells.

(39) TABLE-US-00002 TABLE 2 CI (%) at 24 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 10.sup.−5 Additive mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml TF (bovine) 35 17 29 18 18 15 TF+ 13.5 20.3 35 28.5 10 20.3 (international formulation) TF+ (85:15, 13.3 10.6 29 30 21.6 76 bovine:avian) TF (70:30, 80 47 24 12 30 26.3 bovine:avian) TF (avian) 16 37 47 47 16.1 34.3 None 18 18 18 18 18 18 (spontaneous cell death) (±6%)

(40) TABLE-US-00003 TABLE 3 % Increase in CI (over spontaneous CI) at 24 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 10.sup.−5 Additive mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml TF (bovine) 94 −6 61 0 0 −17 TF+ −25 13 94 58 −44 13 (international formulation) TF+ (85:15, −26 −41 61 67 20 322 bovine:avian) TF (70:30, 344 161 33 −33 67 46 bovine:avian) TF (avian) −11 106 161 161 −11 91 None 0 0 0 0 0 0 (spontaneous cell death) (±6%)

(41) TABLE-US-00004 TABLE 4 CI (%) at 48 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 10.sup.−5 Additive mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml TF (bovine) 19.3 50 54.7 15.3 40.7 11.3 TF+ 23.3 12 17 42 48 62 (international formulation) TF+ (85:15) 48 82.7 96.7 69.4 54 91 TF (70:30) 97 94 99 90 96 91 TF (avian) 68 49 45 35 58 70 None 18 18 18 18 18 18 (spontaneous cell death) (±6%)

(42) TABLE-US-00005 TABLE 5 % Increase in CI (over spontaneous CI) at 48 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 10.sup.−5 Additive mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml TF (bovine) 7 178 204 −15 126 −37 TF+ 29 −33 −6 133 167 244 (international formulation) TF+ (85:15, 167 359 437 286 200 406 bovine:avian) TF (70:30, 439 422 450 400 433 406 bovine:avian) TF (avian) 278 172 150 94 222 289 None 0 0 0 0 0 0 (spontaneous cell death) (±6%)

(43) The data provided in TABLES 2 through 5 confirms that the majority of test samples (i.e., transfer factor-containing compositions) stimulated increased (relative to spontaneous tumor cell death) antitumor and cytotoxic activity of healthy donors' lymphocytes against K-562 tumor cells.

(44) The greatest stimulating effect appears in the 48 hour results, with the most effective range of stimulating concentrations being from about 0.1 mg/ml to about 0.0001 mg/ml. The test samples that included both colostrum-derived transfer factor and egg-derived transfer factor again appear to be the most effective in the given conditions of the experiment, lysing as many as 80-98% of the K-562 tumor cells.

(45) Additionally, the results of TABLE 5 indicate that combinations of different types of transfer factor, particularly the 85:15 ratio of TF+ to egg-derived transfer factor, may be more effective than other courses of therapy for eliminating undesirable cells and pathogens from the body of a treated animal. More specifically, inasmuch as the inventors are aware, in equivalent testing, the best results that could be achieved with interleukin-2 treatment have been 76% cytotoxicity of K-562 tumor cells with a 24 hour incubation (which amounts to a 322% increase over spontaneous deaths of such cells) and an 88% cytotoxicity of K-562 tumor cells with a 48 hour incubation (which amounts to a 389% increase over spontaneous deaths of such cells).

Example 3

(46) Another confirmatory test was conducted to verify the above-stated results and to evaluate the effects of a greater variety of compositions of the present invention on inducing NK and other mononuclear cells to kill K-562 tumor cells. The same protocol described in EXAMPLE 2 was employed in the tests of EXAMPLE 3.

(47) The results of 24 and 48 hour incubation periods for a variety of compositions formulations, each including egg powder and bovine colostrum powder, are listed in TABLES 6 through 9.

(48) TABLE-US-00006 TABLE 6 CI (%) at 24 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10−.sup.4 Bovine:Avian mg/ml mg/ml mg/ml mg/ml mg/ml 85:15 45 29 67.5 28 50 50:50 67.5 23 66 63.5 22.5 30:70 64.6 68.8 39.1 45.6 44 15:85 55.2 28 20.1 20 18.8 None 18 18 18 18 18 (spontaneous cell death) (±6%)

(49) TABLE-US-00007 TABLE 7 % Increase in CI (over spontaneous CI) at 24 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 Bovine:Avian mg/ml mg/ml mg/ml mg/ml mg/ml 85:15 150 61 275 56 178 50:50 275 28 267 253 25 30:70 259 282 117 153 144 15:85 207 56 12 11 4 None 0 0 0 0 0 (spontaneous cell death) (±6%)

(50) TABLE-US-00008 TABLE 8 CI (%) at 48 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 Bovine:Avian mg/ml mg/ml mg/ml mg/ml mg/ml 85:15 46 60 69 67 64 50:50 69 74 74 63 49 30:70 75 83 67 63 45 15:85 77 69 51 42 40 None 18 18 18 18 18 (spontaneous cell death) (±6%)

(51) TABLE-US-00009 TABLE 9 % Increase in CI (over spontaneous CI) at 48 Hours 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 Bovine:Avian mg/ml mg/ml mg/ml mg/ml mg/ml 85:15 156 233 283 272 256 50:50 283 311 311 250 172 30:70 317 361 272 250 150 15:85 328 283 183 133 122 None 0 0 0 0 0 (spontaneous cell death) (±6%)

(52) For the sake of comparison, a whole colostrum sample and a processed transfer factor sample including 100% bovine transfer factor sample (and no avian transfer factor), each including 0.01 mg/ml of transfer factor, were evaluated. At 24 hours, the whole colostrum sample demonstrated a 22% increase in lysis over spontaneous lysis, while the 100% bovine transfer factor sample was responsible for a 103% increase in lysis over spontaneous cell lysis. At 48 hours, the increases in cell lysis were 26% and 203%, respectively.

(53) The data of TABLES 6 through 9, particularly of TABLES 6 and 8, shows that when more colostrum-derived transfer factor is present in a composition according to the present invention (e.g., 85:15), the initial (24 hour test) response may be greater than the response generated by compositions that include less colostrum-derived transfer factor, but does not increase significantly over time (48 hour test).

(54) Compositions (e.g., 50:50 and 30:70) that include more egg-derived transfer factor may provide comparable short term results (24 hour test), but provide much better long term (48 hour test) results.

(55) These results support the theory that combining different types of transfer factors provides a synergistic effect. They also indicate that the proportions of different types of transfer factor in a composition may be tailored to provide a desired result.

Example 4

(56) TABLE-US-00010 TABLE 10 CI (%) 1 10.sup.−1 10.sup.−2 10.sup.−3 10.sup.−4 10.sup.−5 mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml 24 hrs. TF+ (85:15) 13.3 10.6 29 30 21.6 76 (colostrum:egg) 85:15 45 29 67.5 28 50 (colostrum:egg) 48 hrs. TF+ (85:15) 48 82.7 96.7 69.4 54 91 (colostrum:egg) 85:15 46 60 69 67 64 (colostrum:egg)

(57) EXAMPLE 4 compares data obtained in EXAMPLES 2 and 3 above to illustrate that the inclusion of additional components, primarily polysaccharides, in TFP improves the efficiency with which a composition that incorporates teachings of the present invention induces NK and other mononuclear blood cells to kill K-562 tumor cells and, thus, elicits a secondary immune response.

(58) Notably, in the 48 hour test, where polysaccharides were included, cytotoxicity was greater at all dilutions above 0.0001 mg/ml than in comparable compositions that lacked polysaccharides. Thus, polysaccharides are believed to either increase the synergism with which the two or more types of transfer factors act or to provide additional synergism in the elicitation of a secondary immune response.

(59) While the foregoing EXAMPLES and accompanying data demonstrate the effectiveness of compositions that include transfer factor and, in particular, compositions that include two or more different types of transfer factor, in eliciting a T-cell (e.g., NK cell) mediated immune response, transfer factor is also believed to affect the immune system of a treated subject in a number of other ways. For example, and not to limit the scope of the present invention, transfer factor may provide the biochemical benefits disclosed in U.S. patent application Ser. No. 11/122,430, filed May 4, 2005, the disclosure of which is hereby incorporated herein, in its entirety, by this reference. As the benefits of transfer factor are not limited to elicitation of T-cell mediated immune responses, synergy in the biochemical effects of transfer factor may also be recognized when two or more types of transfer factor are combined.

(60) Although the foregoing description contains many specifics, these should not be construed as limiting the scope of the present invention, but merely as providing illustrations of some of the presently preferred embodiments. Similarly, other embodiments may be devised without departing from the spirit or scope of the present invention. Features from different embodiments may be employed in combination. The scope of the invention is, therefore, indicated and limited only by the appended claims and their legal equivalents rather than by the foregoing description. All additions, deletions and modifications to the invention as disclosed herein which fall within the meaning and scope of the claims are to be embraced thereby.