# Patent

## Title
Method of using β-hydroxy-β-methylbutyrate to treat a condition

## Abstract

Disclosed are methods of treating an individual having a condition characterized by an imbalance in type 1 and type 2 cytokine production, wherein the method comprises administering to the individual an amount of β-hydroxy-β-methylbutyrate (HMB) effective to modulate or otherwise cause an increase in the ratio of type 1 to type 2 cytokines, including an increase in the ratio of type 1 to type 2 cytokines without a corresponding increase in type 2 cytokine levels. Also disclosed are methods of using HMB to treat asthma and allergies. The methods of the present invention are based upon the discovery that HMB modulates cytokine production, most typically by increasing type 1 cytokines without a corresponding increase in type 2 cytokines.

## Background
This application is a divisional of U.S. application Ser. No. 11/641,978 filed Dec. 19, 2006, which claims the benefit of U.S. Provisional Patent Application 60/752,253 filed Dec. 19, 2005, the entire disclosures of which are incorporated herein by reference.


TECHNICAL FIELD
The present invention relates to a method of treating individuals with β-hydroxy-β-methylbutyrate (HMB) to modulate cytokine production.
BACKGROUND OF THE INVENTION
Allergies and asthma in the industrialized world have increased in prevalence and severity over recent years. Asthma is now, in fact, the most common chronic illness among children.
Much is known about the pathogenesis of allergies and asthma. Both are immune-based diseases. Both are associated with an imbalance in the relative levels of type-1 and type-2 cytokines in the body. It has been observed that individuals with allergies or asthma have a higher relative ratio of type 2 to type 1 cytokines. It is believed that this skewed ratio then contributes to the pathogenesis of allergies and asthma.
In general, cytokines are cell-produced regulatory proteins that influence, in paracrine or autocrine fashion, cell function. They are produced by immune cells and are therefore categorized by their inducible function and the cell types involved in the response.
Type 1 cytokines, for example, elicit or augment primarily cell-mediated immune responses against pathogens. Type 1 cytokines are involved in inflammatory responses, viral immunity, intracellular parasite immunity and allograft rejection. Type 1 cytokines include interleukin 2 (IL-2), interleukin 12 (IL-12), and interferon γ (IFNγ). Type 1 cytokines can suppress the production of type 2 cytokines.
Type 2 cytokines, by contrast, elicit or augment primarily antibody-mediated immune responses against pathogens. Type 2 cytokines are involved in humoral responses, helminth immunity, and allergic responses. Type 2 cytokines include interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and interleukin 13 (IL-13). Type 2 cytokines can suppress the production of type 1 cytokines.
Given the association between cytokine imbalance in allergies and asthma, it is believed that therapies directed to the normalization of the ratio of type 1 to type 2 cytokine levels will help treat or even prevent such diseases. To that end, it has now been discovered herein that β-hydroxy-β-methylbutyrate (HMB) exposure (in vitro) increases the relative ratio of type-1 to type-2 cytokines in stimulated peripheral blood mononuclear cells (PBMC), thus providing a potential new therapy for treating individuals having or at risk for developing allergies and asthma.
As a commercially available ingredient, HMB is found in a variety of nutritional products. It is also a metabolite of the essential amino acid leucine and is therefore found naturally in the human body. HMB is also found in a variety of plants, including citrus fruits and alfalfa, as well as in catfish. It is also known and used for a variety of purposes, including to build or maintain muscle mass in appropriate individuals and to enhance overall immune function.
To date, however, there have been no reports on the effect of HMB in modulating type 1 and type 2 cytokine production nor any disclosure of the use of HMB to affect cytokine imbalance in treating conditions responsive thereto, including allergies and asthma.
SUMMARY OF THE INVENTION
The methods of the present invention are directed to the modulation of type 1 to type 2 cytokine levels in the body in those individuals afflicted with conditions characterized by a corresponding cytokine imbalance, to thus provide treatment of the underlying condition. Most notable among such conditions are allergies and asthma.
A first embodiment of the present invention is a method of treating an individual having a condition characterized by a relative imbalance of type 1 to type 2 cytokine levels in the body, wherein the method comprises the administration to the individual of an effective amount of β-hydroxy-β-methylbutyrate to thus modulate the imbalance, typically by increasing the relative levels or production of type 1 to type 2 cytokines. The present invention includes those embodiments in which the condition is asthma, allergies, or both.
A second embodiment of the present invention is a method of treating an individual having or at risk for developing allergies, the method comprising the administration to the individual an effective amount of β-hydroxy-β-methylbutyrate (HMB).
A third embodiment of the present invention is a method of treating individuals having or at risk for developing asthma, the method comprising the administration to the individual an effective amount of β-hydroxy-β-methylbutyrate (HMB).
A fourth embodiment of the present invention is a method of treating elderly individuals at risk for developing age-related infections, the method comprising the administration to such individuals an effective amount of β-hydroxy-β-methylbutyrate (HMB).
The present invention is based upon the discovery that peripheral blood mononuclear cells (PBMC) stimulated with the T cell stimulus CD3/CD28 and simultaneously exposed to HMB exhibit a shift in type 1 and type 2 cytokine production, favoring type 1 cytokine production. The shift occurs as HMB exposure increases the production of type 1 cytokines such as interferon-γ (IFNγ), interleukin 12 (IL-12), and interleukin 2 (IL-2), without a corresponding increase in the production of type 2 cytokines such as interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 10 (IL-10), and interleukin 13 (IL-13).



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 summarizes experimental data showing the effect of HMB on type 1 cytokine (IL-2, IL-12, IFNγ, TNFα, GM-CSF) production from peripheral blood mononuclear cells simultaneously stimulated with CD3/CD28 for 24 hours (*p<0.05, paired sample, 2 tailed t-test, comparison to no HMB).
FIG. 2 summarizes experimental data showing the effect of HMB on type 2 cytokine (IL-4, IL-5, IL-10, IL-13) production from peripheral blood mononuclear cells simultaneously stimulated with CD3/CD28 for 24 hours (*p<0.05, paired sample, 2 tailed t-test, comparison to no HMB).
FIG. 3 summarizes experimental data showing the effect of HMB on the ratio cytokine type 1 (IL-2) to type 2 cytokines (IL-4, IL-5, IL-13, IL-10) from peripheral blood mononuclear cells simultaneously stimulated with CD3/CD28 for 24 hours (*p<0.05, paired sample, 2 tailed t-test, comparison to no HMB)
FIG. 4 summarizes experimental data showing the effect of HMB on the ratio cytokine type 1 (IL-12) to type 2 cytokines (IL-4, IL-5, IL-13, IL-10) from peripheral blood mononuclear cells simultaneously stimulated with CD3/CD28 for 24 hours (*p<0.05, paired sample, 2 tailed t-test, comparison to no HMB)
FIG. 5 summarizes experimental data showing the effect of HMB on the ratio cytokine type 1 (IFNγ) to type 2 cytokines (IL-4, IL-5, IL-13, IL-10) from peripheral blood mononuclear cells simultaneously stimulated with CD3/CD28 for 24 hours (*p<0.05, paired sample, 2 tailed t-test, comparison to no HMB)



DETAILED DESCRIPTION
The methods of the present invention comprise the administration of an effective amount of β-hydroxy-β-methylbutyrate (HMB) to an individual in need thereof in the manner and for the purposes described herein. These and other essential or optional elements or features of the methods of the present invention are described in detail hereinafter.
The terms “treating” and “treatment” as used herein, unless otherwise specified, includes preventing a condition, delaying the onset of a condition, reducing the severity of symptoms of a condition, or eliminating some or all of the symptoms of a condition.
The term “ameliorate” as used herein, unless otherwise specified, means to eliminate, delay, or reduce the prevalence or severity of symptoms associated with a condition.
The term “condition” as used herein, unless otherwise specified, includes pathological and non-pathological conditions, all of which are characterized by an aberration or imbalance in the relative amounts of type 1 to type 2 cytokines.
The term “elderly individual” as used herein, unless otherwise specified, means someone more than 60 years old, preferably more than 70 years old.
The term “modulate” as used herein, unless otherwise specified, means to reduce the imbalance (i.e., imbalance associated with a condition) of type 1 to type 2 cytokine levels in the body, or to otherwise increase the ratio of type 1 to type 2 cytokines, including an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels.
All percentages, parts and ratios as used herein are by weight of the total composition, unless otherwise specified. All such weights as they pertain to listed ingredients are based on the active level and, therefore, do not include solvents or by-products that may be included in commercially available materials, unless otherwise specified.
All numerical ranges as used herein, whether or not expressly preceded by the term “about”, are intended and understood to be preceded by that term, unless otherwise specified.
All references to singular characteristics or limitations of the present invention shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made.
All combinations of method or process steps as used herein can be performed in any order, unless otherwise specified or clearly implied to the contrary by the context in which the referenced combination is made.
The methods of the present invention may also be substantially free of any optional or selected essential feature described herein, provided that the remaining method still contains all of the required limitations as described herein.
Embodiments
We now turn to the first embodiment of the present invention. Conditions included within the first embodiment of the present invention include allergy, asthma, solid tumors, cancers including advanced ovarian cancer and melanoma, kidney tumors, and stress, including psychological stress after a burn injury, surgical stress and pre-surgical stress. The methods are especially useful in treating allergy, asthma, or both.
With respect to allergy and asthma, elevated levels of IL-4, a type 2 cytokine, have been associated with the promotion or aggravation of allergy and asthma. Therefore, the first embodiment of the present invention, which is directed to a method of treating an individual having a condition comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, can treat individuals suffering from the symptoms of allergy and asthma because the increase in type 1 cytokines will serve to promote a balanced type 1 to type 2 cytokine profile.
With respect to cancers, including advanced ovarian cancer, studies have shown that the direct injection into the abdominal cavity of the type 1 cytokine IFN-γ may prolong the survival time for women with advanced ovarian cancer. This treatment has been shown to be effective both during the initial chemotherapy as well as after chemotherapy for individuals in whom chemotherapy has failed. Therefore, the first embodiment of the present invention, which is directed to a method of treating an individual having a condition comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, potentiates treatment of individuals with cancer, including advanced ovarian cancer, because the method has been discovered to raise levels of type 1 cytokines, including IFNγ.
With respect to kidney tumors and melanoma, studies have shown that interleukin 2 given as an injection under the skin can treat some kidney tumors and melanoma. When used as a cancer treatment, it is thought that IL-2 strengthens the body's natural defense mechanism and causes some cancer cells to be recognized and eliminated by immune cells. Therefore, the first embodiment of the present invention, which is directed to a method of treating an individual having a condition comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without an increase in type 2 cytokines, potentiates treatment of individuals having kidney tumors or melanoma because the inventors have discovered that the method of the first embodiment of the present invention can raise levels of type 1 cytokines, including IL-2.
With respect to psychological stress after a burn injury, surgical stress and pre-surgical stress, studies have shown that stress increases type 2 and suppresses type 1 cytokine production. The immune system is compromised when individuals experience stress due to the production of type 2 cytokines and the suppression of type 1 cytokines that accompany periods of stress. Therefore, the first embodiment of the present invention, which is directed to a method of treating an individual having a condition comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without an increase in type 2 cytokines, can treat stress after a burn injury, surgical stress and pre-surgical stress because administering to individuals an amount of HMB effective to increase type 1 cytokine levels without increasing type 2 cytokine levels accommodates for the cytokine imbalance associated with stress. The increase in type 1 cytokines promotes a balanced type 1 to type 2 cytokine profile in the individual.
The type 1 cytokines included within the first embodiment of the present invention include interferon-γ, interleukin 2, and interleukin 12. The type 2 cytokines included within the first embodiment of the present invention include interleukin 4, interleukin 5, interleukin 10, and interleukin 13.
Some of the protective functions of IFNγ include inhibition of viral replication, stimulation of macrophages and enhancement of cell surface molecules necessary for self-recognition in an immune response. Additionally, adequate levels of IFNγ are required for protection against infection and disease. IFNγ also antagonizes several actions of type 2 cytokine IL-4 and inhibits the proliferation of IL-4 producing cells. Therefore, the ability to induce production of IFNγ aids in the treatment of individuals with conditions such as those discussed herein. The inventors have discovered that HMB can induce the production of IFNγ without affecting the production of type 2 cytokines and therefore the present method is effective in treating conditions of the type discussed herein.
Some of the protective functions of IL-2 include inducing proliferation of all T cells, activated B cells, and natural killer cell and enhancing killing of tumor cells by the induction of tumoricidal cytokines from T cells and natural killer cells. Adequate levels of IL-2 are also required for protection against infection and disease. Therefore, the ability to induce production of IL-2 aids in the treatment of individuals with conditions such as those discussed herein. The inventors have discovered that administration of HMB induces the production of IL-2 without increasing the type 2 cytokine levels and therefore the present method is effective in treating conditions of the type discussed herein.
While adequate levels of type 2 cytokine IL-4 are also required for protection against infection and disease, elevated levels of IL-4 have been associated with the promotion of allergies, asthma and stress. Therefore, the ability to treat an individual having a condition as described herein is dependent upon both the ability to induce the production of type 1 cytokines such as IFNγ and IL-2, but also the ability to not simultaneously increase the production of type 2 cytokines, and particularly IL-4, as increased levels of IL-4 are known to promote allergies, asthma and stress. The first embodiment of the present invention is directed to a method of treating an individual having a condition wherein the administration of HMB induces the production of IL-2 and IFNγ without a corresponding increase in IL-4 levels.
Another aspect of the first embodiment of the present invention is directed to a method of treating an individual having a condition characterized by an imbalance in type 1 and type 2 cytokines, comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, wherein the amount of HMB administered is an amount effective to ameliorate allergic symptoms. Elevated levels of type 2 cytokine IL-4 are associated with the promotion of allergies. But type 1 cytokines such as IFNγ antagonize several actions of IL-4 and inhibit the proliferation of IL-4 producing cells. Therefore, the present method is capable of ameliorating the symptoms of allergies when the amount of HMB administered to an individual is an effective amount to promote a balanced type 1 to type 2 cytokine profile.
Similarly, the present invention is directed to a method of treating an individual having a condition characterized by an imbalance in type 1 and 2 cytokines, comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, wherein the amount of HMB administered is an amount effective to ameliorate asthmatic symptoms. Elevated levels of type 2 cytokine IL-4 are associated with the promotion of asthma. But type 1 cytokines such as IFNγ antagonize several actions of IL-4 and inhibits the proliferation of IL-4 producing cells. Therefore, the present method is capable of ameliorating the symptoms of asthma when the amount of HMB administered to an individual is an effective amount to promote a balanced type 1 to type 2 cytokine profile.
Still another aspect of the first embodiment of the present invention is directed to a method of treating an individual having a condition characterized by an imbalance in type 1 and type 2 cytokines, comprising administering to the individual an amount of HMB effective to modulate or otherwise cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, wherein the amount of HMB administered is an amount effective to prevent a decrease in FEV1, or the forced expiratory volume in 1 second. Individuals suffering from severe and persistent asthma display a low FEV1 percentage value, while those who suffer from only mild and intermittent asthma display a higher percentage value. Thus, in administering an amount of HMB effective to induce the production of type 1 cytokines without inducing the production of type 2 cytokines in order to decrease the promotion of asthma associated with imbalanced type 1 to type 2 cytokine profiles, the present method is capable of preventing a decrease in FEV1.
Individuals who experience only mild and intermittent symptoms of asthma display a FEV1 value of greater than or equal to 80%. Therefore, another aspect of the first embodiment of the present invention is directed to a method of treating an individual having a condition where cytokine production is induced, comprising administering to the individual an amount of HMB effective to cause an increase in type 1 cytokine levels without a corresponding increase in type 2 cytokine levels, wherein the amount of HMB administered is an amount effective to maintain basal FEV1 above 80%. In altering the imbalanced type 1 to type 2 cytokine profiles associated with asthma through the method of the first embodiment of the present invention, the method is capable of maintaining basal FEV1 above 80%.
We now turn to the second embodiment of the present invention. The present invention includes a method of treating allergy in an individual having or at risk for developing allergy, the method comprising the administration to the individual an amount of β-hydroxy-β-methylbutyrate effective to prevent or ameliorate symptoms of allergies. Individuals who are at risk for allergy include those who are already suffering from allergies and those who are genetically or otherwise predisposed to having allergies.
The term “allergies” as used herein includes hay fever, food allergies, allergic conjunctivitis, atopic dermatitis, inhalant (air born allergens) allergy, and other common allergies. Such allergies are often associated with exposure to allergens such as animal danders, pollens, insect stings or bites, house dust, house dust mites, molds, some drugs, and foods, especially fish, eggs, milk and nuts.
We now turn to the third embodiment of the present invention. The present invention includes a method of treating asthma in an individual having or at risk for asthma, the method comprising the administration to the individual of an amount of β-hydroxy-β-methylbutyrate effective to prevent or ameliorate asthmatic symptoms. Individuals who are at risk for asthma include those who are already suffering from asthma and those who are genetically or otherwise predisposed to having asthma.
We now turn to the fourth embodiment of the present invention. The present invention includes a method of treating elderly individuals at risk of developing age-related infections, including both bacterial and viral infections, respiratory and non-respiratory, the method comprising the administration to such individuals of an amount of β-hydroxy-β-methylbutyrate effective to reduce the risk or prevalence of such infections.
An effective amount of HMB, for the purposes of the methods described herein, most typically ranges from 0.1 g to 10 g, including from 0.5 g to 5.0 g, and also including from 1.0 g to 3.5 g, of HMB per day. The total daily dose may be administered as a single, divided, or continuous (or semi-continuous) dose (e.g., enteral feeding), every day or on selected intermittent days.
The methods of the present invention are preferably directed to oral administration.
Product Forms
The methods of the present invention may be directed to any product form suitable for the safe administration of an effective amount of HMB to the targeted population or selected individual, all in accordance with the methods herein. Such products include pharmaceutical dosage forms (e.g., capsules, tablets, liquids, topicals, etc.) as well as nutritional products.
Nutritional products for use herein further comprise one or more (preferably all) of fat, protein, carbohydrate, minerals, and vitamins. Such products include solids, liquids, powders, and gels.
Non-limiting examples of solid nutritional product forms suitable for use herein include snack and meal replacement products, including those formulated as bars, sticks, cookies or breads or cakes or other baked goods, frozen liquids, candy, breakfast cereals, powders or granulated solids or other particulates, snack chips or bites, and so forth.
Non-limiting examples of liquid nutritional product forms suitable for use herein include snack and meal replacement products such as those formulated as juices or other acidified beverages, milk or soy-based beverages, shakes, coffees, teas, carbonated beverages, non-carbonated beverages, enteral feeding compositions, and so forth. These liquid compositions are most typically formulated as suspensions or emulsions, but can also be formulated in any other suitable form such as solutions, liquid gels, and so forth.
Many different sources and types of proteins, lipids, and carbohydrates are known and can be used in the various nutritional products described herein, provided that the selected nutrients are safe and effective for oral administration and are compatible with the essential and other added ingredients.
Carbohydrates suitable for use in the nutritional products may be simple, complex, or variations or combinations thereof. Non-limiting examples of suitable carbohydrates include hydrolyzed or modified starch or cornstarch, maltodextrin, glucose polymers, sucrose, corn syrup, corn syrup solids, rice-derived carbohydrate, glucose, fructose, lactose, high fructose corn syrup, indigestible oligosaccharides (e.g., fructooligosaccharides), honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), and combinations thereof.
Carbohydrates suitable for use herein also include soluble dietary fiber, non-limiting examples of which include gum arabic, sodium carboxymethyl cellulose, guar gum, citrus pectin, low and high methoxy pectin, oat and barley glucans, carrageenan, psyllium and combinations thereof. Soluble dietary fiber is also suitable as a carbohydrate source herein, non-limiting examples of which include oat hull fiber, pea hull fiber, soy hull fiber, soy cotyledon fiber, sugar beet fiber, cellulose, corn bran, and combinations thereof.
Proteins suitable for use in the nutritional products include hydrolyzed, partially hydrolyzed or non-hydrolyzed proteins or protein sources, and can be derived from any known or otherwise suitable source such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, corn), vegetable (e.g., soy), or combinations thereof. The proteins for use herein can also include, or be entirely or partially replaced by, free amino acids known for use in nutritional products, non-limiting examples of which include tryptophan, glutamine, tyrosine, methionine, cysteine, arginine, and combinations thereof.
Fats suitable for use in the nutritional products include coconut oil, fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil, high oleic safflower oil, MCT oil (medium chain triglycerides), sunflower oil, high oleic sunflower oil, palm and palm kernel oils, palm olein, canola oil, marine oils, cottonseed oils, and combinations thereof.
The concentration or amount of carbohydrate, protein, and carbohydrate in the nutritional compositions of the present invention can vary considerably depending upon the particular product form and the various other formulations and targeted dietary needs. These macronutrients are most typically formulated within any of the caloric ranges (embodiments A, B, or C) described in the following table.












Nutritional Embodiments












Nutrients
A
B
C






Carbohydrate - % total calories
1-98
10-75
30-50



Fat - % total calories
1-98
20-85
35-55



Protein - % total calories
1-98
 5-70
15-35









The nutritional compositions for use herein may further comprise other optional components that may modify the physical, chemical, aesthetic or processing characteristics of the products or serve as pharmaceutical or additional nutritional components when used in the targeted population. Many such optional ingredients are known or otherwise suitable for use in medical food or other nutritional products or pharmaceutical dosage forms and may also be used in the compositions herein, provided that such optional ingredients are safe for oral administration and are compatible with the essential and other ingredients in the selected product form.
Non-limiting examples of such optional ingredients include preservatives, anti-oxidants, emulsifying agents, buffers, additional pharmaceutical actives, additional nutrients as described herein, sweeteners including artificial sweeteners (e.g., saccharine, aspartame, acesulfame K, sucralose) colorants, flavors, thickening agents and stabilizers, emulsifying agents, lubricants, and so forth.
The nutritional compositions for use herein may further comprise any of a variety of other vitamins or related nutrients, non-limiting examples of which include vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12, carotenoids (e.g., beta-carotene, zeaxanthin, lutein, lycopene), niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, salts and derivatives thereof, and combinations thereof.
The nutritional compositions for use herein may further comprise any of a variety of other additional minerals, non-limiting examples of which include calcium, phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, chloride, and combinations thereof.
Experiment
The following experiment is conducted to determine the relationship between HMB exposure and cytokine production.
To induce production of cytokines, PBMCs, isolated from peripheral blood of 10 normal healthy donors, are stimulated for 24 hours with the T cell stimulant CD3/CD28. Cytokine production is analyzed using a Bio-plex Cytokine Assay. The Bio-Plex technology is based on antibody-antigen interactions, wherein fluorescently labeled beads conjugated with antibody directed against the cytokine of interest bind target cytokine to the bead. This bead-cytokine complex is then exposed to a biotinylated detection antibody and a streptavidin-PE (phycoerythrin) reporter molecule. The signal from the reporter molecule is directly proportional to the amount of cytokine present, thus enabling cytokine quantification.
Each of the T cell-derived cytokines quantified in the experiment is described in the following table:












Type 1 cytokines








Interleukin 2
Growth factor for all subpopulations of T cells


(IL-2)
and also promotes the proliferation of activated



B cells


Interleukin 12
Induces the synthesis of IFNγ, IL-2, and


(IL-12)
Tumor necrosis factor α (TNFα) from helper



T cells committed to the production of type 1



cytokines (Th1 cells), promotes the generation of



lymphokine activated killer cells, inhibits the



synthesis of IgE production


Interferon γ
Influences cell mediated mechanisms of cytotoxicity,


(IFNγ)
has antiviral and antiparasitic activities and



inhibits the proliferation of transformed cells


Granulocyte
Stimulates the proliferation and differentiation


Macrophage
of neutrophilic, eosinophilic, and monocytic


colony
lineages and activates the mature forms of these


stimulating
cell types


factor



(GM-CSF)



Tumor necrosis
Induces cytolysis and cytostasis of tumor cells,


factor α
enhances the proliferation of T cells, promotes


(TNFα)
the proliferation and differentiation of B cells



in the presence of IL-2







Type 2 cytokines








Interleukin 4
Promotes the proliferation and differentiation of


(IL-4)
activated B cells


Interleukin 5
Promotes growth and differentiation of eosinophils


(IL-5)



Interleukin 13
Down-modulates macrophage activity, reduces pro-


(IL-13)
inflammatory cytokine production, induces human



monocyte differentiation and B cell differentiation



and proliferation


Interleukin 10
Suppressive cytokine which down-regulates type 1


(IL-10)
cytokine production









A t-test (paired sample, two-tailed) is performed in which a broad range of type 1 and type 2 cytokines are evaluated from cultures with HMB compared to cultures and without HMB.

Significant dose response increases are then observed in the production of the following type 1 cytokines: IL-2 (5 mM and 10 mM HMB), IL-12 (5 mM and 10 mM HMB), and IFNγ, (5 mM and 10 mM HMB); the results of which are summarized in FIG. 1.
Concerning type 2 cytokine production, a significant decrease is seen in IL-10 production following 10 mM exposure to HMB (see FIG. 2), while HMB does not significantly affect the production of GC-CSF, TNFα, IL-4, IL-5, and IL-13. These results are summarized in FIG. 1 and FIG. 2.
The shifts in cytokine production favoring type 1 are summarized in FIGS. 3, 4, and 5. Increases in IL-2 production relative to IL-4 and IL-10 production are demonstrated at HMB concentrations of 5 mM and 10 mM, while increased in IL-2 production relative to IL-5 and IL-13 production are demonstrated at HMB concentrations of 1 mM, 5 mM, and 10 mM (FIG. 3). Increases in IL-12 production relative to IL-4, IL-5, IL-13, and IL-10 are demonstrated at HMB concentrations of 5 mM and 10 mM (FIG. 4). Increases in IFNγ production relative to IL-4 and IL-10 are demonstrated at an HMB concentration of 10 mM, while relative to IL-5 increases are demonstrated at HMB concentrations of 5 mM and 10 mM (FIG. 5)
The data show that HMB exposure increases type 1 cytokine production (IL-2, IL-12, IFNγ) while reducing production of certain type 2 cytokines (IL-10) and not significantly affecting the production of other type 2 cytokines (GC-CSF, TNFα, IL-4, IL-5, IL-13). The net result, therefore, is a shift in type 1 and type 2 cytokine production in favor of type 1 cytokine production.
EXAMPLES
The following examples illustrate specific embodiments of the methods of the present invention, including some nutritional and other product forms suitable for use therein. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible without departing from the spirit and scope of the invention.
The nutritional compositions described below are representative examples of nutritional products suitable for use in the methods of the present invention. Each may be prepared by conventional methods of making nutritional emulsion, some examples of which are described in U.S. Patent Publication 20050215640A1, which description is incorporated herein by reference.










Liquid Nutritional #1 (Weight Gain Formula)










Ingredient
Amt (kg)
Ingredient
Amt (kg)













Water
316
Vitamin DEK
0.04


Ultratrace/trace min.
0.06
premix



premix

Carrageenan
0.03


Potassium chloride
0.072
Soy lecithin
0.6


Na citrate
2.89
Sodium caseinate
15.5


Potassium iodide
0.0001
Calcium caseinate
4.2


Potassium citrate
1.5
Ca HMB
2.6


Corn syrup
7.68
monohydrate



Maltodextrin
53.6
Milk protein isolate
14


Mg phosphate dibasic
0.26
Sardine oil
6.9


Ca phosphate tribasic
0.99
Ascorbic acid
0.12


Magnesium chloride
1.2
KOH 45% Soln
0.13


Sucrose
11.9
Taurine
0.12


Fructooligosaccharide
5.9
Water sol. vit. premix
0.11


Medium chain
2.6
Ascorbyl palmitate
0.03


triglycerides

Choline chloride
0.25


Canola oil
1.5
L-carnitine
0.0681


Soy oil
0.87
Flavor #1
1.6


57% Vitamin A palmitate
0.007
Flavor #2
0.27



















Liquid Nutritional #2 (Low Glycemic Index Formula)











Amt per

Amt per


Ingredient
1,000 kg
Ingredient
1,000 kg














Water
QS
Vitamin C
584
gm












Maltodextrin
56
kg
Potassium chloride
530
gm


Acid casein
41.09
kg
Choline chloride
472.1
gm


Fructose
28
kg
45% KOH soln.
402.5
gm


High oleic
27.2
kg
UTM/TM premix
369.3
gm


safflower oil


K phosphate
333
gm


Maltitol syrup
16
kg
Carnitine
230.5
gm


Maltitol
12.63
kg
Gellan gum
125
gm


Fibersol 2E
8.421
kg
Taurine
100.1
gm


Caseinate
6.043
kg
Vitamin E
99
gm


FOS
4.607
kg
Lutein Esters (5%)
92
gm


Soy
4.3
kg
WSV premix
75.4
gm


polysaccharide


Vit. DEK premix
65.34
gm


Canola oil
3.2
kg
30% Beta carotene
8.9
gm


Tricalcium
2.8
kg
Vitamin A
8.04
gm


phosphate


Pyridoxine HCl
3.7
gm


Mg chloride
2.4
kg
Chromium chloride
1.22
gm


Lecithin
1.6
kg
Folic acid
0.64
gm


Sodium citrate
1.18
kg
Potassium iodide
0.20
gm


Potassium citrate
1.146
kg
Cyanocobalamin
0.013
gm


Sodium hydroxide
1.134
kg
Vitamin C
584
gm


Mg phosphate
1.028
kg





Calcium HMB
5.7
kg





monohydrate







m-inositol
914
gm



















Liquid Nutritional #3 (Pediatric Formula)










Ingredient
per 771 kg














Stock PIF Slurry





High oleic safflower oil
40.7
kg



Soy oil
24.4
kg



MCT oil
16.3
kg



Lecithin
840.2
g



Monoglycerides
840.2
g



Carrageenan
508.9
g



Caseinate
32.8
kg



Stock OSV Blend





DEK premix
83.3
g



Vitamin A
7.1
g



Lutein esters (5%)
92
g



Stock PIW Slurry





Water
530
kg



Caseinate
11.3
kg



Whey protein
11.9
kg



Stock MIN Slurry





Water
18
kg



Cellulose gum
1696
g



Calcium HMB monohydrate
4.4
kg



Magnesium chloride
2.7
kg



Potassium chloride
1.0
kg



Potassium citrate
2.7
kg



Potassium iodide
0.25
g



Dipotassium phosphate
1.45
kg



Final Blend





PIW slurry
251
kg



PIF slurry
53
kg



MIN slurry
12.6
kg



Sodium chloride
127.4
g



Sucrose
77.6
kg



Tricalcium phosphate
2.5
kg



Water
167
kg



Stock WSF Soln





Water
31.7
kg



Potassium citrate
3.74
g



UTM/TM premix
172.2
g



WSV premix
134.1
g



m-inositol
176.7
g



Taurine
145.5
g



L-carnitine
34.92
g



Choline chloride
638.7
g



Stock ascorbic acid soln.





Water
18.6
kg



Ascorbic acid
550.0
g



45% KOH
341
g



Stock vanilla soln.





Water
38.5
kg



Vanilla flavor
4.3
kg



















Nutritional Liquid #4 (Nutritional Supplement)










Ingredient
per 1,000 kg
Ingredient
per 1,000 kg














Water
QS
Magnesium
558
gm












Corn Syrup
33
kg
chloride




Maltodextrin
28
kg
Vanilla Flavor
544
gm


Sucrose
19.4
kg
Sodium Chloride
272
gm


Caseinate
8.7
kg
Carrageenan
227
gm


Calcium HMB
5.7
kg
Choline chloride
218
gm


monohydrate


UTM/TM Premix
165
gm


High Oleic
4.1
kg
Potassium Chloride
146
gm


Safflower Oil


Ascorbic Acid
145
gm


Canola Oil
4.1
kg
Sodium Citrate
119
gm


Soy Protein
3.7
kg
Potassium
104
gm


Whey Protein
3.2
kg
Hydroxide




Caseinate
2.9
kg
Lutein (5%)
46
gm


Corn Oil
2.0
kg
WSV Premix
33
gm


Tricalcium
1.4
kg
Vit DEK Premix
29
gm


Phosphate


Vitamin A
3.7
gm


Potassium Citrate
1.3
kg
Potassium Iodide
86
mcg


Magnesium
952
gm





Phosphate







Lecithin
658
gm



















Liquid Nutritional #5 (Asthma and Allergy formula)











kg per

kg per


Ingredient
1000 kg
Ingredient
1000 kg













Ingredient water
Q.S.
Natural Vitamin E
0.645


Borage oil
61.1
Micronized tri
0.631


Marine oil
53.4
calcium phosphate



Milk protein isolate
30.4
Tocopherol-2
0.600


Sucrose
11.7
antioxidant



Whey protein conc.
8.41
Taurine
0.456


Gum arabic
8.00
Vanilla
0.400


Calcium HMB
5.7
Sucralose 25% sol
0.375


Monohydrate

Zinc Sulfate
0.251


Soy lecithin
4.77
Ascorbyl palmitate
0.143


Cellulose gum
4.00
Sodium chloride
0.143


Potassium citrate
2.64
Acesulfame K
0.0750


Orange Cream Flavor
2.50
Cupric sulfate
0.0177


Ascorbic acid
1.13
FD&C Red #3
0.0150


Turmeric powder
1.00
B carotene 30%
0.00992


Sodium citrate
0.901
Vit. A palmitate
0.00315


KOH 45% solution
0.799
Sodium molybdate
0.000529


Orange Oil
0.750
Sodium selenate
0.000441



















Powder Nutritional #6 (Exercise Formula)











Amount per

Amount per


Ingredient Name
1000 kg
Ingredient Name
1000 kg















Whey Protein
282.051
kg
Potassium
5.128
kg


Concentrate


Chloride




Calcium Caseinate
192.308
kg
Salt
3.205
kg


Maltodextrin
165.416
kg
Xanthan Gum
3.205
kg


Milk Protein
138.782
kg
Choline Bitartrate
2.782
kg


Isolate


41% choline




Dutch Cocoa 10/12
76.932
kg
Acesulfame K
2.718
kg


Sunflower Oil
21.474
kg
Vanilla
1.923
kg


Creamer


Disodium Phosphate




Myoplex Oil
19.231
kg
Anhydrous
1.667
kg


PreBlend


MicroChill WPI
1.282
kg


Chocolate Cream
15.256
kg
Beta Carotene
1.128
kg


Calcium HMB
13.157
kg
1% CWS




monohydrate


Sucralose
692.3
g


Oat Fiber
10.897
kg
Potassium Citrate
641.0
g


Tricalcium
8.526
kg
38% K




Phosphate


Alpha Ketoglutaric
321.0
g


Vitamin Mineral
8.462
kg
Acid




Preblend


Egg Albumin
321.0
g


Dipotassium
8.333
kg
Powder




Phosphate


L-Glutamine
321.0
g


Rich Dark Chocolate
7.051
kg
Taurine
321.0
g


Carrageenan CSM 2
6.474
kg









Working Example I
A 28-year old individual who suffers from seasonal allergies in the spring is given 0.25-1 g of HMB (Nutritional Liquid #5) four times a week for a year. The symptoms of seasonal allergies are reduced the following spring.
Working Example II
A 30-year old white male who normally has four exacerbations of asthma per year is administered 1-10 g of HMB (Nutritional Liquid #5) four times a week for a year. Exacerbation frequency decreases to once a year.
Working Example III
A 45-year old female who has undergone chemotherapy for ovarian cancer is administered 2-10 g of HMB (Nutritional Liquid #1) four times a week for a year. One year later the ovarian cancer has not returned.
Working Example IV
A 50-year old male diagnosed with and has been treated for kidney tumors is administered 750 mg of HMB (capsules) four times a week for a year. Six months later the tumor has not spread to other parts of the individual's body.
Working Example V
A 42-year old female diagnosed with and treated for melanoma is administered 1 g of HMB 4 times ((Nutritional Liquid #1) a week for a year. Six months later the melanoma has not spread to other parts of the individual's body.
Working Example VI
A 37-year old male suffering from severe symptoms of psychological stress as a result of a burn injury is administered 500 mg (Nutritional Liquid #1) of HMB 4 times a week for a year. One year later the symptoms of psychological stress are reduced.
Working Example VII
A 29-year old female suffering from symptoms of surgical stress is administered 200 mg of HMB (Nutritional Liquid #2) 7 times a week for 2 months. Two months later the symptoms of surgical stress are reduced.
Working Example VIII
A 25-year old male suffering from symptoms of pre-surgical stress is administered 200 mg of HMB (capsules) once a day for 3 weeks preceding the individual's scheduled surgery. At the end of the 3 weeks, symptoms of pre-surgical stress are reduced.
Working Example IX
A 24-year old male suffering from moderate persistent asthma is tested to determine the individual's FEV1 percentage value and the value is recorded. The individual is then administered 5-10 g of HMB 4 times ((Nutritional Liquid #5) a week for a year. One year later the individual's FEV1 has not decreased and symptoms of asthma are reduced.
Working Example X
A 33-year old male suffering from mild intermittent asthma is tested to determine the individual's FEV1 percentage value and a value of 83% is recorded. The individual is then administered 1.5-6 g of HMB 4 times (Nutritional Liquid #5) a week for a year. One year later the individual's FEV1 remains above 80% and symptoms of asthma are reduced.
Working Example XI
A 14-year old female with a family history of seasonal allergies shows no signs of suffering from seasonal allergies. The individual is administered 0.1-1.5 g of HMB (Nutritional Liquid #3) once a day for 6 months. Six months later the individual still shows no signs of suffering from seasonal allergies.
Working Example XII
A 16-year old male with a family history of asthma shows no signs of suffering from the symptoms of asthma. The individual is administered 250 mg of HMB (Nutritional Liquid #4) once a day for 6 months. Six months later the individual still shows no signs of suffering from symptoms of asthma.
Working Example XIII
A 72 year old male, after conventional treatment for and recovery from pneumonia, is administered 250 mg of HMB (Nutritional Liquid #1) once a day for 6 months. During those six months, the individual remains free of respiratory tract infections, including any recurrence of pneumonia.
Working Example XIV
A 24-year old male is training for the New York Marathon. During his training period and for 3 months following the event he takes two servings per day of Nutritional formula #6 (containing 1 gram HMB per serving). Contrary to his previous year's experience, he does not experience any respiratory infections during this intense training regimen (such infections reflect the immune suppression that is known to be associated with extreme physical training programs).

## Claims

1. A method of treating an individual having a condition characterized by a higher relative ratio of type 2 cytokines to type 1 cytokines, the method comprising administering to the individual a composition comprising an effective amount of β-hydroxy-β-methylbutyrate to increase the relative ratio of type 1 to type 2 cytokines, wherein said treating consists of at least one of delaying the onset of said condition and reducing the severity of symptoms of said condition, and wherein said condition is selected from the group consisting of allergy and asthma.

2. The method of claim 1 wherein the level of type 1 cytokines is increased by the administration of β-hydroxy-β-methylbutyrate without an increase in type 2 cytokines.

3. The method of claim 1 wherein the effective amount of β-hydroxy-β-methylbutyrate ranges from 0.1 grams to 10 grams per day.

4. The method of claim 3 wherein the effective amount of β-hydroxy-β-methylbutyrate ranges from 0.5 grams to 10 grams per day.

5. The method of claim 4 wherein the effective amount of β-hydroxy-β-methylbutyrate ranges from 0.5 grams to 5.0 grams per day.

6. The method of claim 5 wherein the effective amount of β-hydroxy-β-methylbutyrate ranges from 1.0 grams to 3.5 grams per day.

7. The method of claim 1 wherein the composition further comprises one or more of fat, protein, and carbohydrate.

8. The method of claim 1 wherein the condition is an allergy.

9. The method of claim 8 wherein the amount of β-hydroxy-β-methylbutyrate is effective to ameliorate symptoms of said allergy.

10. The method of claim 8 wherein the allergy is selected from the group consisting of hay fever, food allergies, allergic conjunctivitis, atopic dermatitis, and inhalant allergies.

11. The method of claim 1 wherein the condition is asthma.

12. The method of claim 11 wherein the amount of β-hydroxy-β-methylbutyrate is effective to ameliorate symptoms of said asthma.

13. The method of claim 11 wherein the amount of β-hydroxy-β-methylbutyrate is effective to prevent a decrease in forced expiratory volume in 1 second (FEV1).

14. The method of claim 11 wherein the amount of β-hydroxy-β-methylbutyrate is sufficient to maintain basal forced expiratory volume in 1 second (FEV1) above 80%.

15. The method of claim 1 wherein the type 1 cytokine is selected from the group consisting of interferon-γ, interleukin 2, and interleukin 12.

16. The method of claim 15 wherein the type 1 cytokine is interferon-γ or interleukin 2.

17. The method of claim 1 wherein the type 2 cytokine is selected from the group consisting of interleukin 4, interleukin 5, interleukin 13, and interleukin 10.

18. The method of claim 17 wherein the type 2 cytokine is interleukin 4.

19. The method of claim 1 wherein the composition further comprises a mineral selected from the group consisting of calcium, phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, chloride, and combinations thereof.