U.S. patent application number 17/558467 was filed with the patent office on 2022-04-14 for compositions including different types of transfer factor.
The applicant listed for this patent is 4Life Patents, LLC. Invention is credited to F. Joseph Daugherty, William J. Hennen, David Lisonbee.
Application Number | 20220112256 17/558467 |
Document ID | / |
Family ID | 1000006042181 |
Filed Date | 2022-04-14 |
United States Patent
Application |
20220112256 |
Kind Code |
A1 |
Lisonbee; David ; et
al. |
April 14, 2022 |
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.
Inventors: |
Lisonbee; David; (Orem,
UT) ; Hennen; William J.; (Eagle Mountain, UT)
; Daugherty; F. Joseph; (Omaha, NE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
4Life Patents, LLC |
Sandy |
UT |
US |
|
|
Family ID: |
1000006042181 |
Appl. No.: |
17/558467 |
Filed: |
December 21, 2021 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
16673651 |
Nov 4, 2019 |
11203625 |
|
|
17558467 |
|
|
|
|
11377703 |
Mar 15, 2006 |
10464980 |
|
|
16673651 |
|
|
|
|
PCT/US04/30307 |
Sep 15, 2004 |
|
|
|
11377703 |
|
|
|
|
10663353 |
Sep 15, 2003 |
6866868 |
|
|
PCT/US04/30307 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61J 3/074 20130101;
A61K 35/00 20130101; A61K 35/20 20130101; A61K 35/57 20130101; C07K
14/52 20130101; C07K 14/465 20130101 |
International
Class: |
C07K 14/52 20060101
C07K014/52; A61K 35/20 20060101 A61K035/20; A61K 35/57 20060101
A61K035/57; A61J 3/07 20060101 A61J003/07; C07K 14/465 20060101
C07K014/465 |
Claims
1. A component consisting of: at least one 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 at least one fraction
and/or extract of bovine colostrum is a fraction of bovine
colostrum.
3. The component of claim 2, wherein the at least one fraction of
bovine colostrum is substantially free of fat.
4. The component of claim 3, wherein the at least one 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 at least one fraction
and/or extract of bovine colostrum is an extract of bovine
colostrum.
6. The component of claim 1, wherein the at least one fraction
and/or extract of bovine colostrum is dried bovine colostrum and
the chicken egg yolk is dried chicken egg yolk.
7. The component of claim 1, wherein the chicken egg yolk is a
fraction of chicken egg yolk.
8. The component of claim 1, wherein the chicken egg yolk is an
extract of chicken egg yolk.
9. The component of claim 1, wherein the at least one fraction
and/or extract of bovine colostrum and the chicken egg yolk of the
component are selected, and amounts and relative proportions of the
at least one 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 at least one 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: at least one 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 at least one 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 at least one 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 at least one 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: at least one 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 at least one 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 at least one 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 at least one 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
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 16/673,651, filed Nov. 4, 2019, titled
COMPOSITIONS INCLUDING DIFFERENT TYPES OF TRANSFER FACTOR, now U.S.
Pat. No. 11,203,625, issued Dec. 21, 2021 ("the '651 Application"),
which is a continuation of U.S. patent application Ser. No.
11/377,703, filed Mar. 15, 2006, titled COMPOSITIONS INCLUDING
DIFFERENT TYPES OF TRANSFER FACTOR, now U.S. Pat. No. 10,464,980,
issued Nov. 5, 2019 ("the '703 Application"), which is a
continuation-in-part of PCT international patent application no.
PCT/US2004/030307, filed Sep. 15, 2004 ("the '307 International
Application"), which claims the benefit of the filing date of U.S.
patent application Ser. No. 10/663,353, filed Sep. 15, 2003, now
U.S. Pat. No. 6,866,868, issued Mar. 15, 2005 ("the '353
Application"). The entire disclosures of each of the '651
Application, the '703 Application, the '307 International
Application, and the '353 Application are hereby incorporated
herein.
TECHNICAL FIELD
[0002] The present invention relates generally to compositions
which include transfer factor and, more specifically, to
compositions which include transfer factor from different types of
source animals. The present invention also relates to methods for
making compositions that include different types of transfer factor
and to methods for eliciting or enhancing a T-cell mediated immune
response by the immune system of a subject.
RELATED ART
[0003] Many deadly pathogens are passed to humans from the animal
kingdom. For example, monkeys are the sources of the type I human
immunodeficiency virus (HIV-I), which causes acquired immune
deficiency syndrome (AIDS) and monkeypox, which is similar to
smallpox; ground-dwelling mammals are believed to be the source of
the Ebola virus; fruit bats and pigs are the source of the Nipah
virus; the Hendra virus comes from horses; the virus responsible
for the "Hong Kong Flu" originated in chickens; and wild birds,
especially ducks, are the sources of many of the deadly influenza
viruses. Many diseases also have animal reservoirs. By way of
example, mice carry Hanta virus, rats carry the Black Plague, and
deer carry Lyme disease.
The Immune System
[0004] The immune systems of vertebrates are equipped to recognize
and defend the body from invading pathogenic organisms, such as
parasites, bacteria, fungi, and viruses. Vertebrate immune systems
typically include a cellular component and a noncellular
component.
[0005] The cellular component of an immune system includes the
so-called "lymphocytes," or white blood cells, of which there are
several types. It is the cellular component of a mature immune
system that typically mounts a primary, nonspecific response to
invading pathogens, as well as being involved in a secondary,
specific response to pathogens.
[0006] In the primary, or initial, response to an infection by a
pathogen, white blood cells that are known as phagocytes locate and
attack the invading pathogens. Typically, a phagocyte will
internalize, or "eat" a pathogen, then digest the pathogen. In
addition, white blood cells produce and excrete chemicals in
response to pathogenic infections that are intended to attack the
pathogens or assist in directing the attack on pathogens.
[0007] Only if an infection by invading pathogens continues to
elude the primary immune response is a specific, secondary immune
response to the pathogen needed. As this secondary immune response
is typically delayed, it is also known as "delayed-type
hypersensitivity." A mammal, on its own, will typically not elicit
a secondary immune response to a pathogen until about seven (7) to
about fourteen (14) days after becoming infected with the pathogen.
The secondary immune response is also referred to as an acquired
immunity to specific pathogens. Pathogens have one or more
characteristic proteins, which are referred to as "antigens." In a
secondary immune response, white blood cells known as B
lymphocytes, or "B-cells," and T lymphocytes, or "T-cells," "learn"
to recognize one or more of the antigens of a pathogen. The B-cells
and T-cells work together to generate proteins called "antibodies,"
which are specific for (e.g., configured to bind to or otherwise
"recognize") one or more certain antigens on a pathogen.
[0008] The T-cells are primarily responsible for the secondary, or
delayed-type hypersensitivity, immune response to a pathogen or
antigenic agent. There are three types of T-cells: T-helper cells,
T-suppressor cells, and antigen-specific T-cells, which are also
referred to as cytotoxic (meaning "cell-killing") T-lymphocytes
(CTLs), or T-killer cells or natural killer (NK) cells. The
T-helper and T-suppressor cells, while not specific for certain
antigens, perform conditioning functions (e.g., the inflammation
that typically accompanies an infection) that assist in the removal
of pathogens or antigenic agents from an infected host. The NK
cells, which comprise about ten to about fifteen percent of
circulating lymphocytes, are important mediators of both natural
and adaptive immunity.
[0009] Antibodies, which make up only a part of the noncellular
component of an immune system, recognize specific antigens and,
thus, are said to be "antigen-specific." The generated antibodies
then basically assist the white blood cells in locating and
eliminating the pathogen from the body. Typically, once a white
blood cell has generated an antibody against a pathogen, the white
blood cell and all of its progenitors continue to produce the
antibody. After an infection is eliminated, a small number of
T-cells and B-cells that correspond to the recognized antigens are
retained in a "resting" state. When the corresponding pathogenic or
antigenic agents again infect the host, the "resting" T-cells and
B-cells activate and, within about forty-eight (48) hours, induce a
rapid immune response. By responding in this manner, the immune
system mounts a secondary immune response to a pathogen, the immune
system is said to have a "memory" for that pathogen.
[0010] Mammalian immune systems are also known to produce smaller
proteins, known as "transfer factors," as part of a secondary
immune response to infecting pathogens. Transfer factors are
another noncellular part of a mammalian immune system.
Antigen-specific transfer factors are believed to be structurally
analogous to antibodies, but on a much smaller molecular scale.
Both antigen-specific transfer factors and antibodies include
antigen-specific sites. In addition, both transfer factors and
antibodies include highly conserved regions that interact with
receptor sites on their respective effector cells. In transfer
factor and antibody molecules, a third, "linker," region connects
the antigen-specific sites and the highly conserved regions.
The Role of Transfer Factor in the Immune System
[0011] Transfer factor is a low molecular weight isolate of
lymphocytes. Narrowly, transfer factors may have specificity for
single antigens. U.S. Pat. Nos. 5,840,700 and 5,470,835, both of
which issued to Kirkpatrick et al. (hereinafter collectively
referred to as "the Kirkpatrick Patents"), disclose the isolation
of transfer factors that are specific for certain antigens. More
broadly, "specific" transfer factors have been generated from cell
cultures of monoclonal lymphocytes. Even if these transfer factors
are generated against a single pathogen, they have specificity for
a variety of antigenic sites of that pathogen. Thus, these transfer
factors are said to be "pathogen-specific" rather than
antigen-specific. Similarly, transfer factors that are obtained
from a host that has been infected with a certain pathogen are
pathogen-specific. Although such preparations are often referred to
in the art as being "antigen-specific" due to their ability to
elicit a secondary immune response when a particular antigen is
present, transfer factors having different specificities may also
be present in such preparations. Thus, even the so-called
"antigen-specific," pathogen-specific transfer factor preparations
may be specific for a variety of antigens.
[0012] Additionally, it is believed that antigen-specific and
pathogen-specific transfer factors may cause a host to elicit a
delayed-type hypersensitivity immune response to pathogens or
antigens for which such transfer factor molecules are not specific.
Transfer factor "draws" at least the non-specific T-cells, the
T-inducer and T-suppressor cells, to an infecting pathogen or
antigenic agent to facilitate a secondary, or delayed-type
hypersensitivity, immune response to the infecting pathogen or
antigenic agent.
[0013] Typically, transfer factor includes an isolate of proteins
having molecular weights of less than about 10,000 daltons (D) that
have been obtained from immunologically active mammalian sources.
It is known that transfer factor, when added either in vitro or in
vivo to mammalian immune cell systems, improves or normalizes the
response of the recipient mammalian immune system.
[0014] The immune systems of newborns have typically not developed,
or "matured," enough to effectively defend the newborn from
invading pathogens. Moreover, prior to birth, many mammals are
protected from a wide range of pathogens by their mothers. Thus,
many newborn mammals cannot immediately elicit a secondary response
to a variety of pathogens. Rather, newborn mammals are typically
given secondary immunity to pathogens by their mothers. One way in
which mothers are known to boost the immune systems of newborns is
by providing the newborn with a set of transfer factors. In
mammals, transfer factor is provided by a mother to a newborn in
colostrum, which is typically replaced by the mother's milk after a
day or two. Transfer factor basically transfers the mother's
acquired, specific (i.e., delayed-type hypersensitive) immunity to
the newborn. This transferred immunity typically conditions the
cells of the newborn's immune system to react against pathogens in
an antigen-specific manner, as well as in an antigen- or
pathogen-nonspecific fashion, until the newborn's immune system is
able on its own to defend the newborn from pathogens. Thus, when
transfer factor is present, the immune system of the newborn is
conditioned to react to pathogens with a hypersensitive response,
such as that which occurs with a typical delayed-type
hypersensitivity response. Accordingly, transfer factor is said to
"jump start" the responsiveness of immune systems to pathogens.
[0015] Much of the research involving transfer factor has been
conducted in recent years. Currently, it is believed that transfer
factor is a protein with a length of about forty-four (44) amino
acids. Transfer factor typically has a molecular weight in the
range of about 3,000 to about 5,000 Daltons (Da), or about 3 kDa to
about 5 kDa, but it may be possible for transfer factor molecules
to have molecular weights outside of this range. Transfer factor is
also believed to include three functional fractions, each of which
may include different types of transfer factor molecules: an
inducer fraction; an immune suppressor fraction; and an
antigen-specific fraction. Many in the art believe that transfer
factor also includes a nucleoside portion, which could be connected
to the protein molecule or separate therefrom, that may enhance the
ability of transfer factor to cause a mammalian immune system to
elicit a secondary immune response. The nucleoside portion may be
part of the inducer or suppressor fractions of transfer factor.
[0016] The antigen-specific region of the antigen-specific transfer
factors is believed to comprise about eight (8) to about twelve
(12) amino acids. A second highly-conserved region of about ten
(10) amino acids is thought to be a very high-affinity T-cell
receptor binding region. The remaining amino acids may serve to
link the two active regions or may have additional, as yet
undiscovered properties. The antigen-specific region of a transfer
factor molecule, which is analogous to the known antigen-specific
structure of an antibody, but on a much smaller molecular weight
scale, appears to be hyper-variable and is adapted to recognize a
characteristic protein on one or more pathogens. The inducer and
immune suppressor fractions are believed to impart transfer factor
with its ability to condition the various cells of the immune
system so that the cells are more fully responsive to the
pathogenic stimuli in their environment.
Sources of Noncellular Immune System Components
[0017] Conventionally, transfer factor has been obtained from the
colostrum of milk cows, such as by the method described in U.S.
Pat. No. 4,816,563 to Wilson et al. (hereinafter "Wilson"). While
milk cows typically produce large amounts of colostrum and, thus,
large amounts of transfer factor over a relatively short period of
time, milk cows only produce colostrum for about a day or a
day-and-a-half every year. Thus, milk cows are neither a constant
source of transfer factor nor an efficient source of transfer
factor.
[0018] Transfer factor has also been obtained from a wide variety
of other mammalian sources. For example, in researching transfer
factor, mice have been used as a source for transfer factor.
Antigens are typically introduced subcutaneously into mice, which
are then sacrificed following a delayed-type hypersensitivity
reaction to the antigens. Transfer factor is then obtained from
spleen cells of the mice.
[0019] While different mechanisms are typically used to generate
the production of antibodies, the original source for antibodies
may also be mammalian. For example, monoclonal antibodies may be
obtained by injecting a mouse, a rabbit, or another mammal with an
antigen, obtaining antibody-producing cells from the mammal, then
fusing the antibody-producing cells with immortalized cells to
produce a hybridoma cell line, which will continue to produce the
monoclonal antibodies throughout several generations of cells and,
thus, for long periods of time.
[0020] Antibodies against mammalian pathogens have been obtained
from a wide variety of sources, including mice, rabbits, pigs,
cows, and other mammals. In addition, the pathogens that cause some
human diseases, such as the common cold, are known to originate in
birds. As it has become recognized that avian (i.e., bird) immune
systems and mammalian immune systems are very similar, some
researchers have turned to birds as a source for generating
antibodies.
[0021] Avian antibodies that are specific for pathogens that infect
mammals, or "mammalian pathogens," have been obtained by
introducing antigens into eggs. Alternatively, antibodies may be
present in eggs following exposure of the source animal to
antigens, including antigens of mammalian pathogens. U.S. Pat. No.
5,080,895, issued to Tokoro on Jan. 14, 1992 (hereinafter "the '895
Patent"), discloses a method that includes injecting hens with
pathogens that cause intestinal infectious diseases in neonatal
mammals. The hens then produce antibodies that are specific for
these pathogens, which are present in eggs laid by the hens. The
'895 Patent discloses compositions that include these
pathogen-specific antibodies and use thereof to treat and prevent
intestinal diseases in neonatal piglets and calves. Treatment of
pathogenic infections in mammals with avian antibodies may have
undesirable results, however, since the immune systems of mammals
may respond negatively to the large avian antibody molecules by
eliciting an immune response to the antibodies themselves.
Moreover, as mammalian immune systems do not recognize avian
antibodies as useful for their abilities to recognize certain
pathogens, or the specificities of avian antibodies for antigens of
such pathogens, avian antibodies often do not elicit the desired
immune responses in mammals.
[0022] It is also known that transfer factor may be obtained from
eggs. U.S. Pat. No. 6,468,534 to Hennen et al. (hereinafter
"Hennen") describes a process by which female chickens (i.e., hens)
are exposed to one or more antigens, which results in the
elicitation of an immune response, including a secondary immune
response, by the chickens. As a result of the secondary immune
response, transfer factor molecules are present in the eggs of the
chicken. The eggs may then be processed to provide a product in
which the transfer factor is present. Such a product may take the
form of a spray dried or freeze dried, or lyophilized, egg powder,
and may include all or part of the egg. The egg powder may then be
incorporated directly into gelatin capsules or mixed with other
substances then introduced into gelatin capsules.
[0023] FIG. 2 schematically depicts capsulation equipment of a type
that is currently useful for capsulating egg-derived avian transfer
factor in the form of an egg powder. Capsulation equipment 20
includes a composition supply hopper 24, a feed station 28, and an
auger 26 in communication between each composition supply hopper 24
and feed station 28. Auger 26 transports the whole egg powder from
composition supply hopper 24 to feed station 28.
[0024] When auger 26 operates, it is heated to a temperature that
exceeds the relatively low melting point of cholesterol, from egg
yolk, in the egg powder. The warmed cholesterol is sticky, coating
auger 26, the conduit in communication therewith, and feed station
28, thereby decreasing the efficiency with which capsulation
equipment 20 operates. Consequently, capsulation equipment 20 must
be disassembled and cleaned periodically, which may take a
considerable amount of time (e.g., up to about 8 hours), resulting
in a significant decrease in the productivity of capsulation
equipment 20 and, thus, the number of capsules that may be formed
therewith. Thus, processing of whole egg powder to obtain a
transfer factor-containing product is somewhat undesirable.
[0025] Additionally, compositions which are derived from products
(e.g., eggs or colostrum) from a single source animal typically
only include transfer factor molecules which have specificity to
antigens to which the source animal has been exposed. The
consequence of such limited exposure may be that the effectiveness
of such transfer factor-containing compositions in preventing or
treating certain types of infections or conditions is also
limited.
[0026] Accordingly, there is a need for a composition which is
useful for causing an immune system of a treated subject to elicit
an immune response to a broader array of pathogens, as well as for
a method for improving the efficiency and productivity with which
capsulation and other composition-forming equipment operates.
SUMMARY OF THE INVENTION
[0027] The present invention includes compositions for eliciting
T-cell mediated immune responses in subjects. The composition
includes an active component with transfer factor from at least two
different types of source animals. The term "type," as used herein
with respect to source animals, describes the source animals from
which transfer factor may be obtained and refers to source animals
from different classes (e.g., mammals, birds, reptiles, amphibians,
insects, etc.). The term "type," as used herein, also refers to
source animals from different subclasses, orders (e.g.,
artiodactyls, primates, carnivores, etc.), families (bovine,
hominids, felines, etc.), subfamilies, genuses (e.g., cattle,
humans, domestic cats, etc.), and even species and subspecies. Use
of the term "type" herein with respect to transfer factor denotes
the type of source animal from which the transfer factor was
obtained.
[0028] An exemplary embodiment of the active component of such a
composition includes transfer factor from both mammalian and
nonmammalian source animals, which types of transfer factor are
also referred to herein as "mammalian transfer factor" and
"nonmammalian transfer factor," respectively. By way of nonlimiting
example, the mammalian transfer factor may be included in the
composition as colostrum or a fraction or extract thereof, which
are collectively referred to herein as "colostrum-derived
products," or otherwise, as known in the art (e.g., as a cellular
extract, such as a leukocyte (white blood cell) extract, a splenic
("from the spleen") extract, or the like, etc.). Also by way of
example, the nonmammalian transfer factor of the exemplary
composition may be obtained from an egg or a fraction or extract
thereof, which are also referred to herein as "egg-derived
products." It has been discovered that when different types of
transfer factors are combined and administered to a treated animal
(e.g., a mammal), some synergy occurs.
[0029] When a composition of the present invention includes a
colostrum-derived product and an egg-derived product, both products
may be included in the mixture in amounts (e.g., by weight, by
volume, etc., of the total mixture) that are about equal, or more
of one of the colostrum-derived product and the egg-derived product
than the other. Experimental results show that transfer factor from
source animals that have highly dependent young, such as cows,
induces a relatively quick secondary immune response, with anergy
(i.e., a lack of sensitivity by white blood cells to the transfer
factor molecules) setting in relatively quickly.
[0030] The different types of transfer factor of the active
component may be selected or provided in amounts that are tailored
to cause a treated subject to synergistically elicit a T-cell
mediated immune response. For example, transfer factor from source
animals that have independent young, such as chickens or other
"gallinaceous" birds, does not induce as quick a secondary immune
response, but does provide for a more sustained secondary immune
response. Accordingly, the relative concentrations of
colostrum-derived transfer product and egg-derived product may be
tailored to elicit a secondary immune response that occurs or is
sustained for a particular period of time. As another example of
such synergism, transfer factor from one source may facilitate
elicitation of a cell-mediated immune response against a
corresponding set of pathogens or other antigenic agents, while
transfer factor from another source may cause a treated subject to
elicit a cell-mediated response against another set of pathogens or
other antigenic agents. As a further example, one set of pathogens
against which transfer factor from one source (e.g., from a source
animal that has been exposed to a broad array of pathogens or other
antigenic agents) is most effective may cause a subject to elicit a
broad, or unfocused immune response, while transfer factor from
another source (e.g., a source from a source animal that has been
exposed to a limited number (e.g., only one or a few) pathogens or
other antigenic agents) may cause a subject to elicit a narrow,
focused immune response.
[0031] An active component of such a composition may consist
essentially of the two or more types of transfer factor (including
dialysate or another at least partially purified fraction having an
upper-end molecular weight cutoff of about 10,000 Da), or include
additional components.
[0032] Additional components may include a variety of different
things, such as a portion of a source (e.g., egg, colostrum, cells,
etc.) from which the transfer factor was derived, a supplement,
beneficial microorganisms, and the like.
[0033] If a portion, or extract, of a source of transfer factor is
included in a composition according to the present invention, the
extract may be purified at least partially to remove one or more
components therefrom. By way of nonlimiting example, proteins
(e.g., antibodies and other proteins having molecular weights of
about 160,000 Da or more), fat, casein, cells, or cell debris may
be substantially removed from the extract and, thus, the extract,
or even the composition, may be substantially free of these
components. Allergenic components, including, but not limited to,
some of the components listing immediately above, may also be
separated from the transfer factor from at least one source. Of
course, there is no requirement that any components be
substantially removed from non-transfer factor portions of one or
more sources, or that the non-transfer factor portions of one or
more sources otherwise be purified.
[0034] Supplements are also referred to herein as "supplemental
components." A supplement that may be included in a composition of
the present invention includes, without limitation, one or more
vitamins, minerals, proteins, or natural products (e.g., herbs,
mushrooms, roots, etc.) or extracts thereof. Polysaccharides are
believed to provide further synergy in the effectiveness of a
composition of the present invention in eliciting secondary immune
responses in treated animals. Exemplary polysaccharides are
available in the form of beta-glucans and mushroom extracts (which,
of course, include other components).
[0035] While a composition according to the present invention may
also or alternatively include one or more beneficial
microorganisms, compositions that incorporate teachings of the
present invention may also lack microorganisms and, thus, be
microorganism-free or cell-free.
[0036] According to another aspect, the present invention includes
methods for forming compositions that include two or more types of
transfer factor. One or more of transfer factor (e.g., colostrum,
eggs, cells, tissues, etc.) may be processed to obtain and,
optionally, at least partially purify transfer factor. Such
processing may also be used to obtain, extract, or at least
partially purify other components from the one or more sources. For
example, processes such as those disclosed in Wilson and Hennen,
may be employed. If desired, other components may be included in
the composition.
[0037] In another aspect, the present invention includes a method
for processing or manufacturing an egg-derived product which
includes transfer factor. The inventive method of processing or
manufacture includes mixing a substantially fat-free component,
such as a colostrum-derived product, which may or may not include
transfer factor, with the egg-derived product before or while the
egg-derived product is being introduced into manufacturing or other
processing equipment. Capsulation is one example of a processing or
manufacturing method in which such techniques may be employed.
[0038] Additionally, the present invention includes a method for
reducing the cleaning frequency of manufacturing or other
processing equipment, such as capsulation equipment, used for
processing an egg-derived product. That method includes mixing a
less fatty or substantially fat free substance, such as a
colostrum-derived product, with the egg-derived product before or
during introduction of the egg-derived product into the processing
equipment.
[0039] The present invention also includes methods for treating a
subject. Treatment methods that incorporate teachings of the
present invention include administration of a composition according
to the present invention to a subject. As the composition includes
transfer factor, administration of the composition to the subject
will cause the subject's immune system to elicit a T-cell mediated
immune response or will enhance a T-cell mediated immune response
by the subject's immune system which is already underway.
[0040] Other features and advantages of the present invention will
become apparent to those of ordinary skill in the art through
consideration of the ensuing description, the accompanying
drawings, and the appended claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041] In the drawings, which depict exemplary embodiments of
various aspects of the present invention:
[0042] FIG. 1 depicts an example of the manner in which a
composition that incorporates teachings of the present invention
may be embodied;
[0043] 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
[0044] FIG. 3 schematically illustrates an exemplary test protocol
that was conducted to determine the efficacy of various aspects of
the present invention.
DETAILED DESCRIPTION
[0045] 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.
[0046] 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."
[0047] 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.
[0048] 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).
[0049] 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.
[0050] 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.
[0051] 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.
[0052] 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.
[0053] 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.
[0054] 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.
[0055] 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.
[0056] 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.
[0057] 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.
[0058] 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.
[0059] 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.
[0060] 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.
[0061] 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.
[0062] 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
[0063] 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).
[0064] 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.
[0065] 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.TM. (TF), available from
4Life Research, LLC, of Sandy, Utah, Transfer Factor Plus.TM. (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.
[0066] 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.
[0067] 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
TABLE-US-00001 [0068] TABLE 1 Target Cells Composition 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 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 US 6,196/ 5,543/
4,008/ 8,050/ 4,693/ % increase over flour 458% 485% 306% 389% 362%
% increase over average 380% 270% 178% 438% 244% Bov-Av TFP Intl
5,747/ 4,786/ 3,640/ 7,366/ 4,269/ % increase over flour 424% 417%
277% 355% 328% % increase over average 346% 219% 152% 393% 213%
100% Avian TF 2,553/ 1,860/ 2,483/ 2,985/ 2,183/ % increase over
flour 93% 66% 96% 48% 73% % increase over average 98% 24% 72% 100%
60%
[0069] 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.
[0070] 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.
[0071] 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
[0072] 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
.mu.l of culture medium, were then introduced in 100 .mu.l 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.RTM., as shown at reference character 44.
[0073] 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.degree. C., and incubated for
periods of 24 hours and 48 hours. Each study variant was conducted
in triplicate.
[0074] 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.
[0075] 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.
[0076] 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 .mu.l aliquots of dimethylsulfoxide (DMSO)
were introduced into the wells.
[0077] 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.
[0078] 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:
CI(%)=[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.
TABLE-US-00002 TABLE 2 CI (%) at 24 Hours Additive 1 mg/ml
10.sup.-1 mg/ml 10.sup.-2 mg/ml 10.sup.-3 mg/ml 10.sup.-4 mg/ml
10.sup.-5 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%)
TABLE-US-00003 TABLE 3 % Increase in CI (over spontaneous CI) at 24
Hours Additive 1 mg/ml 10.sup.-1 mg/ml 10.sup.-2 mg/ml 10.sup.-3
mg/ml 10.sup.-4 mg/ml 10.sup.-5 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%)
TABLE-US-00004 TABLE 4 CI (%) at 48 Hours Additive 1 mg/ml
10.sup.-1 mg/ml 10.sup.-2 mg/ml 10.sup.-3 mg/ml 10.sup.-4 mg/ml
10.sup.-5 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%)
TABLE-US-00005 TABLE 5 % Increase in CI (over spontaneous CI) at 48
Hours Additive 1 mg/ml 10.sup.-1 mg/ml 10.sup.-2 mg/ml 10.sup.-3
mg/ml 10.sup.-4 mg/ml 10.sup.-5 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%)
[0079] 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.
[0080] 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.
[0081] 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
[0082] 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.
[0083] 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.
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%)
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%)
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%)
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%)
[0084] 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.
[0085] 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).
[0086] 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.
[0087] 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
TABLE-US-00010 [0088] TABLE 10 CI (%) 1 mg/ml 10.sup.-1 mg/ml
10.sup.-2 mg/ml 10.sup.-3 mg/ml 10.sup.-4 mg/ml 10.sup.-5 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)
[0089] 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.
[0090] 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.
[0091] 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.
[0092] 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.
* * * * *