Mixed allergen compositions and methods of use thereof

Abstract

A mixed allergen composition of two or more different allergens is provided. In some cases, the mixed allergen composition comprises: a nut allergen; an animal allergen; and at least one of: a non-nut plant allergen; a biologic; and a vitamin. Also provided is a method of administering the mixed allergen composition to a subject. The mixed allergen composition can be used for a variety of applications, including health maintenance applications, immune balance applications, intestinal balance applications, immune support applications, health improvement applications, and therapeutic applications.

Description

[0001] This application is a divisional application of CN201680017213.2.

[0002] Cross-references

[0003] This application claims priority pursuant to 35 U.S.C. §119(e) to U.S. Provisional Patent Application Serial No. 62 / 119,014, filed on February 20, 2015, the disclosure of which is incorporated herein by reference.

[0004] introduction

[0005] Allergy is a disorder of the immune system characterized by the occurrence of allergic reactions to normally harmless environmental substances. Allergic reactions are caused by allergens, which can be present in a variety of sources, including but not limited to pollen or other plant components, dust, mold or fungi, food, additives, latex, infusion reactions, animal or bird dander, insect venom, radiocontrast agents, drugs or chemicals. Common allergic reactions include eczema, urticaria, hay fever, asthma and reactions to venom. Mild allergic reactions like hay fever are very common in the population, causing symptoms such as allergic conjunctivitis, itching and runny nose. In some people, severe allergic reactions to environmental or dietary allergens can lead to life-threatening anaphylactic reactions and potential death if not treated. Allergic reactions can occur in three different modes: a) early reactions or acute responses, b) late reactions and c) potential chronic allergic inflammation. The early stage of allergic reaction usually occurs within a few minutes or even seconds after the first allergen exposure, wherein this early stage is also commonly referred to as immediate allergic reaction. In the early stage of allergic reaction, hypersensitivity reaction to allergen (first encounter) causes response in Th2 cells, which are a subgroup of T cells that produce cytokine interleukin-4 (IL-4). Th2 cells interact with B cells (lymphocytes that produce antibodies to antigens) and together with the action of IL-4 stimulate B cells to begin to produce and secrete immunoglobulin E (IgE).

[0006] IgE plays an important role in allergic reactions and allergic reactions. When allergens are introduced, the B cells of the corresponding subjects produce a large amount of IgE. IgE triggers an immune response by binding to receptors present on basophils and mast cells. When activated, these cells release chemical mediators such as histamine and cytokines that cause the characteristic symptoms of allergic reactions.

[0007] Food allergy is an adverse immune response to food allergens such as food proteins. Common food allergens are found in shellfish, peanuts, tree nuts, fish, milk, eggs, soybeans and fresh fruits such as strawberries, mangoes, bananas and apples. Food allergy mediated by immunoglobulin E (IgE) is classified as type I immediate hypersensitivity. These allergic reactions have an acute onset (from a few seconds to an hour) and the accompanying symptoms may include angioedema (soft tissue swelling of the eyelids, face, lips, tongue, throat and organs); urticaria; itching of the mouth, throat, eyes, skin; gastrointestinal symptoms such as nausea, vomiting, diarrhea, stomach cramps or abdominal pain; rhinorrhoea or nasal congestion; wheezing, shortness of breath or difficulty swallowing; and even anaphylaxis, a severe systemic allergic reaction that can lead to death. Gupta et al. (PEDIATRICS, Vol. 128, No. 1, July 1, 2011, pp. e9-e17) demonstrated that 1 in 12 children under the age of 21 is diagnosed by a physician with food allergy. This prevalence is reported to double every 10 years for certain nuts (CDC 2009). In addition, deaths due to fatal food allergies still occur every year. Importantly, more than $24 billion is spent annually on health care / nursing costs for food allergies (Gupta et al., JAMA PEDIATRICS, November 2013, Vol. 167, No. 11). Most of these costs are due to approximately 90,000 emergency room visits in the United States each year due to food-induced allergic symptoms. Summary of the invention

[0008] A mixed allergen composition of two or more different allergens is provided. In some cases, the mixed allergen composition comprises: a nut allergen; an animal allergen; and at least one of: a non-nut plant allergen; a biologic; and a vitamin. Also provided is a method of administering the mixed allergen composition to a subject. The mixed allergen composition can be used for a variety of applications, including health maintenance applications, immune balance applications, intestinal balance applications, immune support applications, health improvement applications, and therapeutic applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 : Th2 cell proliferation (in cells / mL) in ex vivo blood samples from subjects fed different food allergen mixtures. (Note that Figures 1 to 6 For each, for each set of bars on the graph, the order of the test compositions from left to right corresponds to the order of the test compositions listed from top to bottom in the column on the right side of the graph. Figure 1 The bars from left to right are the data for each composition tested from top to bottom in the column listing the test compositions on the right side of the figure.

[0010] Figure 2 : Specific IgG4 levels in plasma from subjects fed different food allergen mixtures (in mg IgG4 / L).

[0011] Figure 3 : Specific IgE levels in plasma from subjects fed different food allergen mixtures (in mg IgE / L).

[0012] Figure 4 : Th2 cell proliferation (in cells / mL) in ex vivo blood samples from subjects fed different food allergen mixtures.

[0013] Figure 5 : Specific IgG4 levels in plasma from subjects fed different food allergen mixtures (in mg IgG4 / L).

[0014] Figure 6 : Specific IgE levels in plasma from subjects fed a food allergen mixture (in mgIgE / L).

[0015] Figure 7 : T cell proliferation in response to gluten, insulin, tetanus, and bacterial flagellin in ex vivo blood samples from subjects fed a mixture of different food allergens. DETAILED DESCRIPTION

[0016] A mixed allergen composition of two or more different allergens is provided. In some cases, the mixed allergen composition comprises: a nut allergen; an animal allergen; and at least one of the following: a non-nut plant allergen; a biologic; and a vitamin. Also provided is a method of administering the mixed allergen composition to a subject. The mixed allergen composition can be used for a variety of applications, including health maintenance applications, immune balance applications, intestinal balance applications, immune support applications, health improvement applications, and therapeutic applications.

[0017] Before describing the present invention method and composition, it should be understood that the present invention is not limited to the specific method or composition described, because the present invention method or composition can certainly vary. It should also be understood that the terminology used herein is only used to describe specific embodiments and is not intended to be limiting, because the scope of the present invention will only be limited by the appended claims.

[0018] If a numerical range is provided, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed (to one tenth of the lower limit unit, unless the context clearly indicates otherwise). Each smaller range between any given value or intermediate value in a given range and any other given value or intermediate value in the given range is included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the range, and any one, none or all of the two end values ​​are included in each range in the smaller range. It is also covered by the present invention, but any specially excluded value in the given range is excluded. When a given range includes one or both of the end values, the scope excluding any one or both of the end values ​​they include is also included in the present invention.

[0019] Unless otherwise indicated, the meaning of all scientific and technological terms used herein is consistent with what those of ordinary skill in the art to which the present invention belongs generally understand. Although any method and material similar or equivalent to the methods and materials described herein can be used to practice or test the present invention, some potential and preferred methods and materials are now described herein. All publications mentioned herein are incorporated herein by reference to disclose and describe methods and / or materials in conjunction with these cited publications. It should be understood that when there is a conflict, the present disclosure will replace any disclosure of the incorporated publication.

[0020] As will be apparent to those skilled in the art after reading this disclosure, the various embodiments described and illustrated herein each have discrete components and features that can be easily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Any described method can be performed in the order of events described or in any other order that is logically possible.

[0021] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "the peptide" includes reference to one or more peptides and equivalents thereof known to those skilled in the art, such as polypeptides, and so forth.

[0022] The publications discussed herein are provided only because their publication dates precede the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to take precedence over such publications because of prior invention. In addition, the publication dates provided may differ from the actual publication dates, and independent confirmation of the actual publication dates may be required.

[0023] Mixed allergen composition

[0024] As outlined above, aspects of the present invention include mixed allergen compositions. By mixed allergen composition is meant a composition comprising two or more different allergens, wherein any two given allergens are different if they are different from each other, for example, they are compounds described by different chemical formulae or compositions described by different components (e.g., constituent compounds) and / or their amounts. The number of different allergens in the composition may vary as desired. In certain embodiments, the mixed allergen composition comprises 2 or more different allergens, such as 3 or more allergens, 4 or more allergens, 5 or more allergens, 6 or more allergens, 7 or more allergens, 8 or more allergens, 9 or more allergens, 10 or more allergens, 15 or more allergens, 20 or more allergens, 25 or more allergens, 30 or more allergens, 40 or more allergens, 50 or more allergens, 75 or more allergens, 100 or more allergens, wherein in some cases the number of different allergens in a given composition is 100 or less, such as 75 or less, including 50 or less, like 25 or less, 15 or less, including 10 or less.

[0025] The allergens present in the composition may vary, wherein in some cases the allergens present in the composition are allergens that can induce allergic reactions in susceptible subjects. Allergens include any antigens that trigger a specific IgE response, or their active derivatives. Antigens include any substances that can stimulate antibody production and can specifically bind to them. Antigenic determinants or epitopes are antigenic sites on molecules. Allergens themselves may have little or no intrinsic toxicity, but due to their ability to trigger IgE-related immune responses, and after subsequent exposure, due to their ability to trigger IgE-dependent and / or T-cell-dependent hypersensitivity reactions, they can cause pathological conditions. As such, "allergens" include any substances that can stimulate typical hypersensitivity reactions in atopic subjects. Allergens that may be present in a given mixed allergen composition include any substances present in various different sources (such as, but not limited to: food, drugs, spices, plants, environments or biological systems (such as prokaryotic or eukaryotic cells or viruses)), as well as chemical allergens.

[0026] Allergens of interest include nut allergens. Nut allergens are allergens that include one or more compounds present in nuts (e.g., dried fruits that include an edible kernel or flesh enclosed in a woody or leathery shell). Nut allergens of interest include, but are not limited to, peanut allergens such as rAra h 1, rAra h 2, rAra h 3, rAra h 8PR-10, rAra h 9LTP, peanut complete allergen (the phrase "complete allergen" as used herein refers to all possible antigenic components of a given food protein), etc.; Brazil nut allergens such as rBer e 1, Brazil nut complete allergen, etc.; hazelnut allergens such as rCor a 1PR-10, rCor a 8LTP, hazelnut complete allergen, nCor a 9, rCor a 14, etc.; walnut allergens such as rJug r 1, rJug r 3LTP, walnut complete allergen, etc.; cashew allergens; pistachio allergens, such as pistachio component allergens, pistachio complete allergens, etc.; pecan allergens, such as pecan component allergens, pecan complete allergens, etc.; tree nut component allergen packages, such as one or more allergens from cashews, walnuts, hazelnuts, Brazil nuts; etc.

[0027] Allergens of interest include animal allergens. Animal allergens are allergens that include one or more compounds present in animals, including both vertebrates and invertebrates. Vertebrate allergens that may be present in the mixed allergen composition include avian allergens such as egg allergens, such as nGal d 1 ovomucoid, nGal d 2 ovalbumin, nGal d 3 conalbumin, egg white complete allergen, etc.; mammalian allergens, such as milk allergens, such as nBos d 4 α-lactalbumin, nBos d 5 β-lactoglobulin, nBos d 8 casein, nBos d lactoferrin, milk complete allergen, etc.; fish allergens, such as rCyp c 1, rGad c 1, cod complete allergen, whitefish allergen, pink salmon allergen, etc. Invertebrate allergens that may be present in the mixed allergen composition include: crustacean allergens, such as shrimp allergens, such as rPen a 1 tropomyosin, shrimp complete allergen, etc.; insect allergens, such as bee sting allergens, wasp sting allergens, mosquito bite allergens, etc.; and the like.

[0028] Allergens of interest include non-nut plant allergens, ie plant allergens that are not nut allergens.Plant allergens are allergens that include one or more compounds present in plants. The plant allergens of concern include: wheat allergens, such as rTri a 19ω-5 gliadin, wheat complete allergen, wheat alcohol-soluble protein, rTri a 14LTP, etc.; kiwi fruit allergens, such as rAct d 8PR-10, kiwi fruit complete allergen, etc.; celery allergens, such as rApi g1.01PR-10, rPhl p 12, celery complete allergen, CCD MUXF3 from bromelain, etc.; soybean allergens, such as rGly m 4PR-10, soybean complete allergen, nGly m 5β-conglycinin, nGly m 6 soybean globulin, etc.; stone fruit allergens, such as f419, f420, f421, f95, f242, o214 rPru p 1PR-10, rPru p 3LTP, stone fruit primary complete allergen, CCD from bromelain, etc. MUXF3, etc.; oat allergens, such as oat component allergens, oat complete allergens, etc.; sesame allergens, such as sesame seed component allergens, sesame seed complete allergens, etc.

[0029] Other types of allergens that may be present in the mixed allergen composition include, but are not limited to: non-food animal allergens, such as cat or dog fur and dander, cockroach calyx, dust mite excrement, etc.; drug allergens, such as penicillin, sulfonamides, salicylates, local anesthetics; mold spore allergens, latex allergens; metal allergens; plant pollen allergens, such as grasses - ryegrass, timothy, weeds - ragweed, plantain, nettle, mugwort, quinoa, Oxalis, trees - birch, alder, hazel, hornbeam, horse chestnut, willow, poplar, sycamore, tilia, olive; and the like.

[0030] The amount of a given allergen in a mixed allergen composition can vary as desired. In some cases, the amount of a given allergen is in the range of 5 to 15,000 mg, such as 10 to 10,000 mg (including 15 to 5,000 mg). The weight percentage of a given allergen in a mixed allergen composition can vary, ranging from 15 to 99.9 weight %, such as 25 to 65 weight %. A mixed allergen composition can be a unit dose composition, and so-called unit dose composition means that it is present in a composition configured to be administered to a subject as a single dose, which may or may not be a part of a dosing regimen consisting of two or more unit doses administered to a subject in a given time period. Although the mass of a given unit dose may vary, in some cases the unit dose has a mass ranging from 300 mg to 20 g, such as 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg (1 g), 1.5 g, 2 g, 3 g, 4 g, 5 g, 10 g, 15 g, 20 g, and any value therebetween. In certain embodiments, any two of the mixed allergens are present in equal parts (e.g., in a 1:1 ratio) such that each allergen is present in the composition in equal weights. In such embodiments where the mixed allergen composition comprises three different allergens, the three allergens are present in a 1:1:1 ratio. For example, a mixed allergen composition may include equal parts (e.g., a ratio of 1:1:1, etc.) of the following allergens: walnuts, hazelnuts, shrimp, salmon, eggs, milk, peanuts, cashews, almonds, and wheat (e.g., about 30 mg of each protein; about 90 mg of each protein; or about 300 mg of each protein).

[0031] In some embodiments, the mixed allergen composition comprises nut allergens and animal allergens; and at least one of the following: non-nut plant allergens; biologics; and vitamins. As such, in some embodiments, the mixed allergen composition comprises nut allergens, animal allergens, and non-nut plant allergens. In such embodiments, the mixed allergen composition may further comprise biologics or vitamins or both biologics and vitamins. In some embodiments, the mixed allergen composition comprises nut allergens, animal allergens, and further comprises biologics. In these embodiments, the mixed allergen composition may further comprise vitamins. In some embodiments, the mixed allergen composition comprises nut allergens, animal allergens, and further comprises vitamins. In these embodiments, the mixed allergen composition may further comprise biologics.

[0032] Biological agents can vary, and include both probiotics and prebiotics. Probiotics are generally live eukaryotic or prokaryotic organisms with beneficial properties when given to a subject. In one aspect, probiotics supplement the existing microbial flora in a subject. Thus, probiotics are live microorganisms that can confer health benefits to a host subject. Probiotics can be cultures of microorganisms or provided in dietary supplements, or can be lyophilized and reconstituted before use. Prebiotics are agents that are beneficial to or confer growth, maintenance and / or beneficial properties of a subject's microbial flora. Prebiotics can include oligosaccharides and soluble or insoluble fiber substances. Examples of probiotics include, but are not limited to, species of Lactobacillus spp., Escherichia spp., Bacillus spp., Bifidobacterium spp., Saccharomyces spp., and Streptococcus spp. Specific probiotics that may be present in the mixed allergen composition include Lactobacillus species, such as Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus casei Shirota, Lactobacillus casei immunitass, Lactobacillus johnsonii, Lactococcus lactis, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus (lactobacillus GG), Lactobacillus salivarius and Lactobacillus helvetirus. The probiotic microorganism can be naturally occurring, attenuated or genetically modified to introduce new traits or change existing traits. In one embodiment, the probiotic has been genetically modified to introduce an allergen gene or its part or fragment, which is expressed to produce a recombinant microorganism that releases the allergen or its antigenic fragment or exposes the subject's immune system to the allergen or its antigenic fragment. Thus, probiotics and allergens can be combined into a single component of the mixed allergen composition. When present, the amount of the biological agent in the mixed allergen composition can vary. In some cases, the biological agent is present in an amount ranging from 1.5 to 99.9 % by weight, such as 10 to 25 % by weight.

[0033] The mixed allergen composition may contain one or more vitamins as desired. The vitamins that may be present in the composition include, but are not limited to: vitamin A, such as in an amount ranging from 1 to 35,000 IU; vitamin C, such as in an amount ranging from 1 to about 1,000 mg; vitamin D, such as in an amount ranging from 1 to 4,000 IU; vitamin E, such as in an amount ranging from 1 to 450 IU; vitamin K, such as in an amount ranging from 1 to 250 mcg; vitamin B-1 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-2 (riboflavin), such as in an amount ranging from 1 to 17 mg; vitamin B-2 (cobalamin), such as in an amount ranging from 1 to 17 mg; vitamin B-3 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-4 (riboflavin), such as in an amount ranging from 1 to 17 mg; vitamin B-5 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-6 (riboflavin), such as in an amount ranging from 1 to 17 mg; vitamin B-7 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-8 (riboflavin), such as in an amount ranging from 1 to 15 mg; vitamin B-9 (riboflavin), such as in an amount ranging from 1 to 15 mg; vitamin B-10 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-11 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-12 (riboflavin), such as in an amount ranging from 1 to 15 mg; vitamin B-13 (thiamine), such as in an amount ranging from 1 to 15 mg; vitamin B-14 (riboflavin), such as in an amount ranging from 1 to Vitamin B-3 (niacin), such as in an amount ranging from 1 to 200 mg; Vitamin B-5 (pantothenic acid), such as in an amount ranging from 1 to 100 mg; Vitamin B-6 (pyridoxine), such as in an amount ranging from 1 to 30 mg; Vitamin B-9 (folic acid), such as in an amount ranging from 1 to 4,000 mcg; Vitamin B-12 (cobalamin), such as in an amount ranging from 1 to 250 mcg; Vitamin H (biotin), such as in an amount ranging from 1 to 1,000 mcg vitamin H (biotin); etc.; and combinations thereof.

[0034] The mixed allergen composition of the present invention can exist in different configurations. In some cases, the mixed allergen composition exists in a solid configuration, for example as a powder. When present as a powder, the size of the particles constituting the powder can vary, in some cases ranging from 0.1 to 1000 microns, such as 1 to 500 microns.

[0035] Also provided is a physiologically acceptable composition comprising the mixed allergen composition and a physiologically acceptable delivery vehicle. The mixed allergen composition can be incorporated into various preparations for administration to a subject. More specifically, the mixed allergen composition can be formulated into a physiologically acceptable composition by combining with a suitable physiologically acceptable carrier or diluent, and can be formulated into a preparation in solid, semisolid, liquid or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols and topical compositions. The preparation can be designed for use via a variety of different approaches (including oral, buccal, sublingual, rectal, parenteral, intraperitoneal, intradermal, transdermal, intratracheal and other approaches).

[0036] The physiological composition may be in a form suitable for oral use, for example as a food, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs, gums, etc. Compositions intended for oral use may be prepared according to any convenient procedures for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from sweeteners, flavoring agents, coloring agents and preservatives in order to provide a palatable preparation.

[0037] Food preparations of interest include mixed allergen compositions in combination with food delivery vehicles. By food delivery vehicle is meant a delivery vehicle for nutrients that are eaten, drunk, or otherwise ingested into the body to sustain life, provide energy, promote growth, etc. Examples of food delivery vehicles of interest include, but are not limited to, infant formula, baby food, potato chips, biscuits, bread, spreads, cream, yogurt, liquid beverages, chocolate-containing products, candy, ice cream, cereals, coffee, etc.

[0038] Also concerned about as oral preparations is food additives. If the oral preparation is provided as a food additive, the food additive may also include one or more of a sweetener, a stabilizer, a flavoring agent or a coloring agent. According to the oral preparation of the present disclosure, it can be provided in the form of a sugar-coated tablet or lozenge, a pill, a gelatin capsule or a syrup. Oral preparations can be provided as bulk samples, such as containers (which can be measured by a subject) containing multiple doses of powder form, or in unit dosage form, such as pills, pouches (pouch), single-use containers, etc. Provide.

[0039] Tablets may contain active ingredients mixed with non-toxic physiologically acceptable excipients suitable for making tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents or disintegrants, for example, corn starch or alginic acid; binders, such as starch, gelatin or gum arabic; and lubricants, for example, magnesium stearate, stearic acid or talc. Tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release over a longer period of time. For example, time-delay materials such as monostearate or distearate may be used. They may also be coated by the techniques described in U.S. Patent Nos. 4,256,108, 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

[0040] Formulations for oral use may also be presented as hard gelatin capsules in which the mixed allergen component is mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules in which the mixed allergen component is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin or olive oil).

[0041] The aqueous suspension contains the mixed allergens mixed with excipients suitable for making aqueous suspensions. Such excipients may include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic; dispersants or wetting agents may be naturally occurring phospholipids such as lecithin, or condensation products of alkylene oxides with fatty acids such as polyoxyethylene stearate, or condensation products of ethylene oxide with long chain fatty alcohols such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides such as polyoxyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate; one or more coloring agents; one or more flavoring agents; and one or more sweetening agents, such as sucrose, saccharin or aspartame.

[0042] Oily suspensions can be prepared by suspending the active ingredient in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (such as liquid paraffin). The oily suspension may contain a thickening agent, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners (such as those described above) and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant (such as ascorbic acid).

[0043] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide the active ingredient in admixture with a dispersant or wetting agent, a suspending agent and one or more preservatives. Examples of suitable dispersants or wetting agents and suspending agents are those already mentioned above. Additional excipients such as sweeteners, flavorings and coloring agents may also be present.

[0044] The physiologically acceptable compositions of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil (e.g., olive oil or peanut oil), or a mineral oil (e.g., liquid paraffin) or a mixture of these substances. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin; and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion can also contain sweeteners and flavorings.

[0045] Syrups and elixirs can be prepared using sweeteners such as glycerol, propylene glycol, sorbitol or sucrose. These preparations may also contain demulcents, preservatives and flavoring agents and coloring agents. The pharmaceutical composition may be in the form of a sterile water suspension for injection or an oil suspension. The suspension may be prepared using suitable dispersants or wetting agents and suspending agents as described above according to known techniques. Sterile injection preparations may also be sterile injection solutions or suspensions in non-toxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils are routinely used as solvents or suspending media. For this purpose, any mild fixed oil may be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid may be used to prepare injections.

[0046] The mixed allergen components can be formulated into preparations for injection by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent, such as vegetable oil or other similar oils, synthetic fatty acid glycerides, esters of higher fatty acids or propylene glycol; and if necessary, together with conventional additives such as solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers and preservatives.

[0047] The mixed allergen components may be used in aerosol formulations to be administered via inhalation. The compounds of the present invention may be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen and the like.

[0048] In addition, the mixed allergen composition can be made into suppositories by mixing with various bases such as emulsifying bases or water-soluble bases. The compounds of the present invention can also be administered via suppositories rectum. Suppositories can include vehicles such as cocoa butter, carbowax and polyethylene glycol, which melt at body temperature and solidify at room temperature.

[0049] Also of interest are topical compositions, such as those in which the mixed allergen composition is combined with a topical delivery vehicle component. The topical delivery vehicle component of the delivery composition of the present invention may vary as desired, wherein the specific composition of a given delivery vehicle component will depend at least in part on the properties of the specific composition. Delivery compositions of interest include liquid preparations such as lotions (liquids containing insoluble substances in the form of suspensions or emulsions, intended for external application, including spray lotions) and aqueous solutions; semisolid preparations such as gels (in which the dispersed phase has been combined with a dispersion medium to produce a semisolid substance such as a jelly colloid), creams (soft solids or thick liquids) and ointments (soft, oily preparations); and solid preparations such as topical patches. As such, delivery vehicle compositions of interest include, but are not limited to, oil-in-water (O / W) and water-in-oil (W / O) emulsions, milks, lotions, creams, ointments, gels, serums, powders, masks, packs, sprays, aerosols, or sticks.

[0050] The amount of the mixed allergen composition that can be combined with a carrier material to prepare a single dosage form will vary depending on the host being treated and the specific mode of administration. For example, a formulation intended for oral administration to a human may contain 0.5 mg to 5 g of a mixed allergen composition, which is compounded with an appropriate and convenient amount of carrier material, which may be about 5% to about 95% of the total composition. However, it should be understood that the specific dosage level for any particular patient will depend on a variety of factors, including age, body weight, overall health, sex, diet, time of administration, route of administration, excretion rate, concurrent use of drugs, and the severity of the specific disease undergoing therapy. As such, unit dosage forms such as syrups, elixirs, and suspensions for oral or rectal administration may be provided, wherein each dosage unit, for example, a teaspoonful, a tablespoonful, a tablet, or a suppository contains a predetermined amount of a composition containing one or more inhibitors. Similarly, a unit dosage form for injection or intravenous administration may include the inhibitor in the composition as a solution in sterile water, saline, or another pharmaceutically acceptable carrier. As described above, "unit dosage forms" include physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a mixed allergen composition calculated sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle.

[0051] method

[0052] Aspects of the present invention also include methods of administering a mixed allergen composition (e.g., a mixed allergen composition as described above) to a subject. The route of administration used in a given method may vary, for example, depending on the properties of the mixed allergen composition. As discussed above, a physiologically acceptable composition comprising a mixed allergen composition may be formulated for delivery to a subject using a variety of different routes of administration, such as, but not limited to, oral administration, buccal administration, sublingual administration, rectal administration, parenteral administration, intraperitoneal administration, intradermal administration, transdermal administration, intratracheal administration, etc. As such, aspects of the present method may include administering a mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen (e.g., as described above) to a subject orally, buccal, sublingual, rectal, parenteral, intraperitoneal, intradermal, transdermal, intratracheal, etc.

[0053] The methods described herein can be used for a variety of different types of subjects, i.e., animals, wherein the animals are generally "mammals" or "mammals," wherein these terms are used in a broad sense to describe organisms within the class Mammalia, including the order Carnivora (e.g., dogs and cats), the order Rodentia (e.g., mice, guinea pigs, and rats), the order Lagomorpha (e.g., rabbits), and the order Primates (e.g., humans, chimpanzees, and monkeys). In some embodiments of the invention, the subject is a human. The subject can be an infant, a teenager, or an adult, wherein in some cases the subject can be a pregnant adult, e.g., as described in more detail below. As such, in certain embodiments, the subject is an infant less than 1 year old, while in other embodiments the subject is older, e.g., 1 year old or older, 5 years old or older, etc., and includes adults.

[0054] The dosing regimen may vary as desired and may depend on many different factors, such as the purpose of administration, the age of the subject, the condition of the subject, the nature of the physiologically acceptable composition, etc. In certain embodiments, the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition is administered to the subject once per hour, once per few hours (e.g., every 2, 3, 4, 6 hours), once per day, once per week, once per two weeks, once per month, once per two months, once per quarter, once per half year, once per year, etc., for a treatment time period, which may also vary, wherein in some cases the treatment time period is 1 day or longer, 1 week or longer, 2 weeks or longer, 1 month or longer, 3 months or longer, 6 months or longer, 1 year or longer, 2 years or longer, 3 years or longer, 5 years or longer, 10 years or longer, etc., up to the life span of the subject.

[0055] In some embodiments, the method is a method for increasing the immune health of the subject. Because the methods of these embodiments are methods for increasing the immune health of the subject, the embodiments of such methods result in the subject being better able to have a healthy immune response to a given attack. Immune health or immune balance can be characterized as a state in which there is no inflammatory imbalance in the body. For example, when IgE decreases and / or IgG4 increases and / or Th2 decreases in plasma (e.g., as described in more detail below in the "Experiments" section and the accompanying drawings). The amplitude of immune health enhancement can vary, wherein in some cases the amplitude is 2 times or more, such as 5 times or more, including 10 times or more, for example compared to a suitable control.

[0056] In some embodiments, the method is a method for increasing the intestinal health of the subject. Because the method of these embodiments is a method for increasing the intestinal health of the subject, the embodiments of these methods cause the subject to have an enhanced healthy microbial flora to maintain or improve the resilience of the microbial flora, for example, by reducing the number or colonization of pathogenic bacteria or viruses and by maintaining and improving intestinal integrity and barrier function. Intestinal health or intestinal balance can be characterized as a state in which there is no inflammatory imbalance in the human body. For example, when T cell proliferation is reduced (for example, as described in more detail below in the "experimental" section and the accompanying drawings). The amplitude of intestinal health enhancement can vary, wherein in some cases the amplitude is 2 times or higher, such as 5 times or higher, including 10 times or higher, for example, compared to a suitable control.

[0057] In some embodiments, the method is a method for enhancing the health of the subject or maintaining the immune balance of the subject. Because the method of these embodiments is a method for enhancing the physical and mental health of the subject, the embodiments of these methods result in the quality or state of the subject's body and mind being healthy. Health status can be characterized as a physical state in which there is no immune imbalance problem (e.g., less IgE, more IgG4, less cellular inflammation (e.g., as described in more detail below in the "Experiments" section and the accompanying drawings). The amplitude of physical and mental health enhancement can vary, wherein in some cases the amplitude is 2 times or more, such as 5 times or more, including 10 times or more, for example compared to a suitable control.

[0058] In some embodiments, the method is a method for at least reducing the potential of a subject to develop an immune-mediated disorder (such as an immune-mediated inflammatory disease disorder). The so-called at least reducing the potential of a subject to develop an immune-mediated disorder means that the probability of a subject developing an immune-mediated disorder is reduced, such as a subject developing an immune-mediated disorder. For example, in order to determine the risk reduction, if 100 different individuals apply the composition, 20% or more subjects will show that their immune markers (such as IgE) are reduced when compared with a control group. The amplitude of the potential reduction can vary, wherein in some cases the amplitude is 2 times or higher, such as 5 times or higher, including 10 times or higher, for example, compared with a suitable control. In some cases, the method is a method for preventing a subject from developing an immune-mediated disorder. As such, the method of the present invention includes administering the composition of the present invention to a subject who is not aware of or does not have an immune-mediated disorder. Although the subject may not suffer from or may not know that he or she suffers from the immune-mediated disorder, the subject may be a subject suspected of having a risk of developing an immune-mediated disorder or a subject known to be at risk of developing an immune-mediated disorder.

[0059] Aspects of the invention also include methods of treating an immune-mediated disorder in a subject. By treatment is meant the improvement of one or more symptoms associated with a disease condition (e.g., an immune-mediated disorder) afflicting the subject, wherein the improvement is used in a broad sense to refer to at least a decrease in the magnitude of a parameter (e.g., a symptom) associated with the pathological condition being treated, etc. As such, treatment also includes wherein the pathological condition or at least a symptom associated therewith is completely inhibited, e.g., prevented from occurring, or is stopped, e.g., terminated, such that the subject no longer suffers from the pathological condition, or at least no longer has symptoms characteristic of the pathological condition.

[0060] Treatment can also be embodied in the form of a surrogate marker for regulating the disease condition. For example, if the target disease is an allergy, such as described below, Th2 cell proliferation may be reduced, such as measured using the assay described in the experimental section below. The amplitude of Th2 cell proliferation reduction can vary, and can be 1.2 times to 10 times in some cases, such as 2 times to 4 times in the scope. If the target disease is an allergy, such as described below, the specific IgG4 level can increase, such as measured using the assay described in the "experimental" section below. The amplitude of the IgG4 level increase can vary, and can be 1.2 times to 100 times in some cases, such as 2 times to 6 times in the scope. If the target disease is an allergy, such as described below, the specific IgE level can be reduced, such as measured using the assay described in the "experimental" section below. The amplitude of the IgE level reduction can vary, and can be 1.1 times to 7 times in some cases, such as 2 times to 6 times in the scope.

[0061] As outlined above, immune-mediated disorders that are targets of the methods of the present invention include immune-mediated inflammatory disorders, wherein such disorders include, but are not limited to, disorders characterized by common inflammatory pathways that lead to inflammation, and which can be caused or triggered by a dysregulation of the normal immune response. Examples of immune-mediated inflammatory disorders include, but are not limited to, allergies, autoimmune diseases, ankylosing spondylitis, psoriasis, psoriatic arthritis, Behcet's disease, arthritis, inflammatory bowel disease (IBD), cardiovascular disease, neuromuscular disease, and infectious diseases, among others.

[0062] In some cases, the target immune-mediated condition is an allergy. The target allergy can vary widely, wherein in some cases the target allergy is a food allergy, a drug allergy, an environmental allergy, an animal allergy, and an insect and / or bee allergy. As such, aspects of the present invention include methods for reducing the risk of a subject developing an allergy. In certain embodiments, the method causes the subject to have a reduced risk of developing a food allergy to a food allergen that is not present in the mixed allergen composition administered to the subject. For example, the composition administered can provide the subject with allergen protection for nut proteins present in the formulation and nut proteins not present in the formulation. In certain embodiments, the use of a mixed formulation is compared to the use of a formulation with only a single allergen, and the risk of the reduction of the first food allergen achieved is reduced to a greater extent. For example, feeding a mixed allergen formulation as described herein can result in a lower risk of developing an allergy to the allergen than when the formulation only contains the single allergen (e.g., with respect to Th2 / IgE / IgG4 analysis, as shown below). In some cases, this phenomenon is referred to as "synergism" (e.g., cashews synergize with pistachios, walnuts synergize with pecans, shrimp synergizes with lobsters (and other crustaceans), and vice versa). If the method is a method of treating an allergy in a subject, as reviewed above, the method may at least result in an improvement in one or more symptoms associated with the allergy (e.g., symptoms as described above in the introduction). Allergy symptoms that may be alleviated include, but are not limited to, eczema, asthma, atopic dermatitis, bronchospasm, cough, rhinorrhoea, angioedema, hypermotility, urticaria, pruritus, fatigue, bradycardia, and / or hypotension. The magnitude of the reduction in symptoms may vary, with the magnitude being 2-fold or more, such as 5-fold or more, including 10-fold or more, for example, compared to a suitable control in some cases. In some cases, treating an allergy results in the subject being cured of the allergy, such that the subject no longer suffers from the allergy. In some embodiments of the allergy treatment method, the method comprises administering to a subject a mixed allergen composition comprising nut allergens; animal allergens; and at least one of: non-nut plant allergens, biologics; and vitamins, such as those described above.

[0063] In some cases, the method is a method for treating an eosinophil disorder in a subject. Eosinophil disorder is characterized by the presence of eosinophils higher than the normal number in various parts of the body. Eosinophil disorders of interest include but are not limited to: eosinophilic esophagitis (esophagus); eosinophilic gastritis (stomach); eosinophilic enteritis (small intestine); eosinophilic colitis (large intestine); high eosinophilic syndrome (blood and any organ) and the like. In some cases the method is a method for reducing the development of eosinophilic disorders in a subject. The amplitude of risk reduction can vary, wherein in some cases the amplitude is 2 times or higher, such as 5 times or higher, including 10 times or higher, for example compared to a suitable control.

[0064] In some cases the method is a method of treating inflammation in a subject, wherein inflammation can be a symptom of a variety of different disease conditions. Disease conditions wherein inflammation can be treated according to embodiments of the present invention include, but are not limited to, celiac disease, multiple sclerosis, inflammatory bowel disease, eosinophilic disease, allergies, food allergies, and the like.

[0065] Aspects of the present invention also include methods for reducing the potential of a fetus or nursing child to develop an immune-mediated disorder (such as an immune-mediated inflammatory disease disorder, such as an allergy, such as the allergy described above). The so-called at least reducing the potential of a fetus or nursing child to develop an immune-mediated disorder means that the probability of a fetus or nursing child developing the immune-mediated disorder is reduced, so that the risk of a fetus or nursing child developing an immune-mediated disorder is reduced. The amplitude of the potential reduction can vary, wherein in some cases the amplitude is 2 times or higher, such as 5 times or higher, including 10 times or higher, for example, compared with a suitable control. In some cases, the method is a method for preventing a fetus or nursing child from developing an immune-mediated disorder. As such, the method of the present invention includes administering the composition of the present invention to a fetus or nursing child who is not aware of an immune-mediated disorder or does not suffer from an immune-mediated disorder. Although the fetus or nursing infant may not have the immune-mediated disorder or may not be known to have the immune-mediated disorder, the fetus or nursing infant may be a subject suspected of being at risk of developing the immune-mediated disorder or known to be at risk of developing the immune-mediated disorder.

[0066] In such an embodiment, the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition is administered to the mother of a fetus or a nursing child. For example, the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition can be administered to a mother pregnant with a fetus. In these cases, any convenient dosing regimen (e.g., as described above) can be used, starting from any convenient time during pregnancy (e.g., at the beginning of mid-term pregnancy, at the beginning of late pregnancy, etc.) the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition is administered to the mother. If the method is a method for reducing the risk of nursing children, the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition can be administered to the mother of the nursing child who breastfeeds. In these cases, any convenient dosing regimen (e.g., as described above) can be used, starting from any convenient time during breastfeeding (e.g., at the beginning of lactation, 1 week after the beginning of lactation, etc.) the mixed allergen composition or a physiologically acceptable composition comprising the mixed allergen composition is administered to the nursing mother.

[0067] The following examples are offered by way of illustration and not by way of limitation.

[0068] experiment

[0069] I. Mixed allergen assay

[0070] A. Materials and Methods

[0071] 1. Allergen treatment

[0072] For each allergen or allergen mixture listed below (allergen group), 5 subjects from each of five different age groups were fed a total of 300 mg of the allergen or allergen mixture daily for one year. The only exceptions were for 3-fold formula (i.e., dose) 1 and 10-fold formula (i.e., dose) 1, where 900 mg and 3,000 mg of the formula were fed daily to the subjects, respectively. The allergen mixture was formulated at a ratio of 1:1. Thus, for an allergen mixture with 2 allergens, the mixture would contain 150 mg of each allergen. Five subjects were included in each age group who were not fed allergens (the "untreated" or "NT group" listed at the end). The five age groups were: 4 to 6 months, 7 to 12 months, 1 to 3 years, 3 to 5 years, 5 years or older. The selection of subjects was not based on observation or suspicion of having a food allergy (or other allergic) condition, so the group of subjects tested included those who might have or had a tendency to develop a food allergy as well as those who did not have such a tendency. The total number of subjects was 450 (18 different treatments (including NT), 5 age groups per treatment, 5 subjects per age group).

[0073] Allergen or mixture of allergens

[0074] 1. Cashews: Finely ground cashews were purchased from "Wellbee's", Spring Valley, NY 10977.

[0075] 2. Cashews and pistachios: Finely ground cashews were purchased from Wellbee's, Spring Valley, NY 10977. Pistachio powder was purchased from nuts.com, Cranford, New Jersey 07016.

[0076] 3. Walnuts: Roasted walnuts were purchased from "Holmquist Hazelnut Orchards" in Lynden WA 98264.

[0077] 4. Walnuts and pecans: Roasted walnuts were purchased from "Holmquist Hazelnut Orchards" in Lynden, WA 98264. Ground pecans were purchased from "King Arthur Flour" in Norwich, Vermont, 05055.

[0078] 5. Formula 1: Equal parts of walnuts, hazelnuts, shrimp, salmon, eggs, milk, peanuts, cashews, almonds and wheat (30 mg protein each)

[0079] 6.3x Formula 1: Equal parts walnuts, hazelnuts, shrimp, salmon, eggs, milk, peanuts, cashews, almonds and wheat (90 mg protein each)

[0080] 7. 10x Formula 1: Equal parts of walnuts, hazelnuts, shrimp, salmon, eggs, milk, peanuts, cashews, almonds and wheat (300 mg protein each)

[0081] 8. Untreated control (NT)

[0082] 9. Cod: Pacific cod, purchased from "Seattle Seafoods" in Bellevue, WA 98008.

[0083] 10. Cod and pink salmon: Pacific cod and sockeye salmon were purchased from "Seattle Seafoods" in Bellevue, WA 98008.

[0084] 11. Shrimp: White shrimp, purchased from "Seattle Seafoods" in Bellevue, WA 98008.

[0085] 12. Shrimp and crab: White shrimp, purchased from "Seattle Seafoods" in Bellevue, WA 98008.

[0086] 13. Peanuts: Defatted peanuts were purchased from "Byrd Mill Company" in Ashland, VA 23005.

[0087] 14. Peanuts and soybeans: Defatted peanuts were purchased from Byrd Mill Company, Ashland, VA 23005. Stone-ground soybeans were purchased from Bob's Red Mill, Milwaukie, Oregon 97222.

[0088] 15. Hazelnuts: Natural hazelnuts purchased from "Holmquist Hazelnut Orchards" in Lynden WA 98264.

[0089] 16. Almonds: blanched almond flour, purchased from "Honeyville" in Rancho Cucamonga, CA 91730.

[0090] 17. Milk: Organic nonfat dry milk powder, purchased from "Now Foods", Bloomingdale, IL 60108.

[0091] 18. Egg: Powdered egg whites with sodium lauryl sulfate as an anti-caking agent were purchased from Honeyville Food Products, Honeyville, Utah 84314.

[0092] 2. Stimulation and Counting of Th2 Cell Subsets

[0093] PBMC from experimenter is labeled with CFSE, and feeds their same food allergen or food allergen mixture (Byrd Mill) with 100 μ g / mL or cultivates 7 days with anti-CD3 / CD28 (in order to test non-specific proliferation ability) to identify T cell subsets.On the 7th day, wash cells and together with Live / Dead staining (Invitrogen), surface CD4, CD25, CD127, D45RO, CD45RA, CD40L and CD69 and intracellular Foxp3 and IL-10 staining (referring to the T cell flow cytometry method below).Th2 cell is defined as the cell (CFSE10) that propagates in response to food allergen and is CD41IL-4, IL-13 cell.Also by separating CD40L and CD69 double positive cells after antigen stimulation, identify the T cell of antigen induction.

[0094] 3. T cell flow cytometry

[0095] Cells were fixed with Lyse / Fix PhosFlow buffer (BD Biosciences). For intracellular staining, fixed cells were permeabilized with Perm buffer III (BD Biosciences) at 48°C for 30 min and then stained for 20 min at 48°C. Flow cytometry was performed using an LSRII flow cytometer (BD Biosciences). Live cells were identified using Live / Dead probes (Invitrogen). Antibodies against surface CD3 (UCHT1), CD4 (SK3), CD25 (4E3), CD127 (SB199), CD45RO (UCHL1), CD45RA (HI100), CD62L (DREG-56), CCR4 (1G1), and CCR8 were purchased from BD Biosciences according to the manufacturer's recommended protocol; antibodies against CCR7, CD69, and CD40L, as well as antibodies against intracellular IL-10 (JES3-19F1), intracellular IL-4 (MP4-25D2), and intracellular IL-13 (JES10-52A2) were purchased from BD Biosciences; antibodies against Helios (22F6) were purchased from BioLegend; anti-CD49b was purchased from BioLegend; and anti-CD49b was purchased from R&D The phenotype of T cells was detected and stained with anti-LAG3 from Invitrogen Systems (Minneapolis, Minn); and an antibody against Foxp3 (150D) purchased from BioLegend.

[0096] 4. Antibody Titer Measurement

[0097] Total and allergen-specific blood IgE and IgG4 levels were measured in all subjects using the standard ImmunoCAP assay (Phadia, Uppsala, Sweden) at the clinical laboratory of Stanford Hospital and Clinics.

[0098] B. Results

[0099] 1. Th2 cell proliferation

[0100] PBMCs from each subject in allergen groups 1 to 14 were stimulated ex vivo for 7 days with the allergen or allergen mixture fed to the subject and analyzed for Th2 cell proliferation (as described in the "Methods" section above). It should be noted that for the 3-fold Formulation 1 group and the 10-fold Formulation 1 group, PBMCs were stimulated ex vivo with 3-fold and 10-fold allergen amounts, respectively (i.e., 300 μg / mL and 1000 μg / mL, respectively).

[0101] like Figure 1 As shown in Figure 1, subjects in allergen groups 1 to 4 and 9 to 14 showed a certain degree of reduction in Th2 proliferation in response to allergen stimulation compared to the untreated control (group 8; baseline-NT). However, the Th2 cell proliferation in PBMCs from subjects in allergen groups 5 to 7 (fed increasing amounts of formula 1) (these subjects were fed the formula containing 10 individual allergens as described above) had a significantly reduced Th2 cell proliferation compared to the control group (NT) as well as all other antigen groups (1 to 4 and 9-14). This phenomenon was seen in all subjects in all allergy groups and at all ages tested.

[0102] 2. Immunoglobulin Analysis

[0103] In addition to the above Th2 proliferation assay, plasma from subjects in allergen groups 1 to 18 was also analyzed (using the standard methods described above) for the presence or absence of IgG4 and IgE antibodies, the former being an indicator of a non-allergic or non-inflammatory (or anti-allergic or non-inflammatory) state, and the latter being an indicator of a pro-allergic or pro-inflammatory state.

[0104] like Figure 2 As shown in the Table 1, subjects in allergen groups 1 to 4 and 9 to 18 showed a moderate increase in IgG4 levels compared to the baseline group (group 8; NT). However, allergen groups 5 to 7 (fed with increasing amounts of formula 1) showed significantly increased levels of IgG4 compared to both the baseline group and all other antigen groups (1 to 4 and 8 to 18). This result is consistent with the results of Th2 cell proliferation as described above. Specifically, increased IgG4 levels and reduced Th2 cell proliferation in plasma are indicators of a reduced allergic or inflammatory state or a non-allergic or non-inflammatory state in a subject.

[0105] like Figure 3As shown in, subjects in allergen groups 1 to 4 and 9 to 17 showed significantly higher IgE levels in plasma compared to allergen groups 5 to 7 (fed with increasing amounts of formula 1). In fact, the IgE levels in allergen groups 5 to 7 were below the detection level of the assay. This result is consistent with the results of Th2 cell proliferation and IgG4 levels as described above. Specifically, low IgE levels and increased IgG4 levels and reduced Th2 cell proliferation in plasma are indicators of a reduced allergic or inflammatory state or a non-allergic or non-inflammatory state in a subject.

[0106] It is obvious from the above results that continuous feeding of a complex mixture of food allergens to very young subjects (e.g., in the form of Formula 1 above) can preventively protect subjects from developing allergic or inflammatory immune features to various antigens. In other words, the process can induce a non-allergic or non-inflammatory (or anti-allergic or anti-inflammatory) state in the subject. Of particular interest in the results shown herein is the evidence that feeding antigens in a complex mixture can provide extensive protection from developing allergic reactions / inflammation, which is superior to single allergen formulations or even dual allergen formulations that target a single allergen (e.g., as in Allergen Groups 1, 3, 11, 13, and 15 to 18 above) or multiple related allergens (e.g., as in Allergen Groups 2, 4, 10, 12, and 14 above).

[0107] It is shown above that feeding a subject with a complex antigen mixture will not only prevent the development of allergic reactions and / or inflammation to allergens / antigens in the complex mixture, but also prevent the development of allergic reactions and / or inflammation to allergens or antigens not present in the complex mixture. In essence, feeding a complex food antigen mixture can produce a general anti-allergic state or anti-inflammatory state in a subject, which can broadly prevent the development of allergic reactions or inflammatory states, even to allergens / antigens that have not been fed to the subject.

[0108] II. Mixed allergen + probiotics and / or vitamins assay

[0109] A. Mixed Allergen Compositions

[0110] exist Figure 4-7 Use a 1:1 blend of:

[0111] 1 dietary supplement blend: 1 probiotic (specifically 300 mg of each). The probiotic in this case is Lactobacillus from Culturelle brand; or

[0112] 1 dietary supplement blend: 1 vitamin (specifically 300 mg of each)

[0113] The vitamins in this case are Poly Vi Sol brand (for children); or

[0114] 1 dietary supplement blend: 1 vitamin: 1 probiotic (specifically 300 mg of each)

[0115] The pH 2 mixture was incubated in vinegar at room temperature for 40 minutes before use.

[0116] The baked mixture was heated at 350°F for 40 minutes.

[0117] The control group received no dietary supplement.

[0118] B. Th2 cell proliferation

[0119] like Figure 4 As shown in , PBMCs from each subject in allergen groups 1 to 14 were stimulated ex vivo for 7 days with allergens or allergen mixtures, or with probiotics, or with vitamins, or under certain conditions at different temperatures or pH, and Th2 cell proliferation was analyzed (as described in the "Methods" section above).

[0120] like Figure 4 As shown in the results, samples from subjects using the mixture or using the mixture under different conditions (i.e. baked goods, skin creams, fruit juices) showed a similar reduction in Th2 proliferation. This phenomenon was seen in all subjects in all allergic groups and at all tested ages.

[0121] C. Immunoglobulin Analysis

[0122] In addition to the above Th2 proliferation assay, plasma from subjects in the allergen group was also analyzed (using the standard methods described above) for the presence or absence of IgG4 and IgE antibodies, the former being an indicator of a non-allergic or non-inflammatory (or anti-allergic or anti-inflammatory) state, and the latter being an indicator of a pro-allergic or pro-inflammatory state.

[0123] like Figure 5 and Figure 6 As shown in , subjects showed modest increases in IgG4 levels and significantly higher IgE levels in plasma compared to controls. Figure 6 As shown in , no changes in IgE levels were observed in response to the different tested mixed allergen compositions. Similar effects were seen when the mixtures were used or when the mixtures were used under different conditions.

[0124] DT cell proliferation

[0125] T cell proliferation was performed (see Methods above) and Figure 7The data are presented in . The data show anti-inflammatory properties against gluten (i.e., a celiac agent) as well as insulin (a diabetes agent) and bacterial flagellin (an inflammatory bowel disease agent). This demonstrates that the mixture may also reduce inflammatory states involved in diseases like celiac disease, diabetes, and inflammatory bowel disease.

[0126] Notwithstanding the accompanying terms, this disclosure also includes the following terms:

[0127] 1. A mixed allergen composition, comprising:

[0128] Nut allergens;

[0129] Animal allergens; and

[0130] At least one of the following:

[0131] Non-nut plant allergens

[0132] Biological agents; and

[0133] Vitamins.

[0134] 2. A mixed allergen composition according to clause 1, wherein the composition comprises non-nut allergens.

[0135] 3. A mixed allergen composition according to clause 1, wherein the composition comprises a biological agent.

[0136] 4. A mixed allergen composition according to clause 3, wherein the biological agent comprises a probiotic.

[0137] 5. A mixed allergen composition according to clause 3, wherein the biologic agent comprises a prebiotic.

[0138] 6. A mixed allergen composition according to clause 1, wherein the composition comprises vitamins.

[0139] 7. A mixed allergen composition according to clause 1, wherein the composition comprises non-nut plant allergens, a biologic agent and vitamins.

[0140] 8. A mixed allergen composition according to any of the preceding clauses, wherein the nut allergen is selected from peanut allergen, Brazil nut allergen, hazelnut allergen, walnut allergen, pistachio allergen, pecan allergen and cashew allergen and combinations thereof.

[0141] 9. The mixed allergen composition according to any one of the preceding clauses, wherein the animal allergen comprises a vertebrate allergen, an invertebrate allergen or a combination thereof.

[0142] 10. The mixed allergen composition according to clause 9, wherein the animal allergen is selected from the group consisting of egg allergens, milk allergens, fish allergens and shrimp allergens and combinations thereof.

[0143] 11. A mixed allergen composition according to any one of the preceding clauses, wherein the non-nut plant allergen is selected from wheat allergen, kiwi fruit allergen, celery allergen, soy allergen, oat allergen, sesame allergen and stone fruit allergen and combinations thereof.

[0144] 12. A mixed allergen composition according to any one of clauses 1 to 11 wherein the amount of each allergen present in the composition ranges from 15 wt% to 99.9 wt%.

[0145] 13. A mixed allergen composition according to clause 12, wherein each allergen is present in the composition in the same weight.

[0146] 14. A mixed allergen composition according to any one of the preceding clauses, wherein the composition is a solid.

[0147] 15. The mixed allergen composition according to clause 14, wherein the composition is a powder.

[0148] 16. A physiologically acceptable composition comprising:

[0149] (a) a mixed allergen composition according to any one of clauses 1 to 15, and

[0150] (b) a physiologically acceptable delivery vehicle.

[0151] 17. A physiologically acceptable composition according to Clause 16, wherein the physiologically acceptable delivery vehicle is an ingestible delivery vehicle.

[0152] 18. A physiologically acceptable composition according to Clause 16, wherein the physiologically acceptable delivery vehicle is a topical delivery vehicle.

[0153] 19. The physiologically acceptable composition according to Clause 16, wherein the physiologically acceptable delivery vehicle is an injectable delivery vehicle.

[0154] 20. A physiologically acceptable composition according to any one of the preceding clauses, wherein the mixed allergen composition is present in the physiologically acceptable composition in an amount ranging from 15 to 99.9% by weight.

[0155] 21. A method comprising:

[0156] administering a mixed allergen composition to a subject, wherein when the method is a method of treating allergy in a subject, the mixed allergen composition is a composition according to any one of clauses 1 to 20.

[0157] 22. The method according to clause 21, wherein the method is a method of increasing the immune health of the subject.

[0158] 23. The method according to clause 21, wherein the method is a method of increasing the intestinal health of the subject.

[0159] 24. The method according to clause 21, wherein the method is a method of enhancing the physical and mental health of the subject.

[0160] 25. The method according to clause 21, wherein the method is a method of at least reducing the potential of the subject to develop an immune-mediated disorder.

[0161] 26. The method according to clause 25, wherein the method is a method of preventing the subject from developing an immune-mediated disorder.

[0162] 27. The method according to clause 25 or 26, wherein the immune-mediated disorder is an allergic disorder.

[0163] 28. The method according to clause 21, wherein the method is a method of treating an immune-mediated disorder in the subject.

[0164] 29. The method according to clause 28, wherein the immune-mediated disorder is an allergic disorder.

[0165] 30. The method according to clause 28 or 29, wherein the method results in improvement of at least one symptom of the immune-mediated disorder.

[0166] 31. A method according to any of the preceding clauses, wherein the subject is a mammal.

[0167] 32. The method of clause 31, wherein the mammal is a human.

[0168] 33. The method according to clause 31 or 32, wherein the subject is an infant.

[0169] 34. A method according to clause 31 or 32, wherein the subject is a teenager.

[0170] 35. The method according to clause 31 or 32, wherein the subject is an adult.

[0171] 36. The method of clause 35, wherein the subject is pregnant or nursing a fetus.

[0172] 37. The method according to clause 36, wherein the method is a method of at least reducing the potential of the fetus or nursing infant to develop an immune-mediated disorder.

[0173] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to those skilled in the art in light of the teachings of this invention that certain changes and modifications may be made therein without departing from the spirit or scope of the appended claims.

[0174] Therefore, the foregoing only illustrates the principle of the present invention. It should be understood that those skilled in the art will be able to design various combinations that embody the principle of the present invention and are included in the spirit and scope of the present invention, although they are not explicitly described or shown here. In addition, all embodiments and conditional language described here are mainly intended to help readers understand the principle of the present invention and the concept that the inventor contributes to promote this area, and should be understood as not being limited to the embodiments and conditions specifically described in this way. Moreover, all statements describing the principles, aspects and embodiments of the present invention and its specific examples here are intended to cover their structural and functional equivalents. In addition, such equivalents are also intended to include currently known equivalents and equivalents developed in the future, that is, no matter how the structure is, but all perform the elements of any development of the same function. The scope of the present invention is therefore not intended to be limited to the exemplary embodiments shown and described here. On the contrary, the spirit and scope of the present invention are embodied by the appended claims.

Claims

1. A mixed allergen composition comprising: Nut allergens, including peanut allergen, almond allergen, hazelnut allergen, walnut allergen, and cashew allergen; Animal allergens, including egg allergens, milk allergens, salmon allergens, and shrimp allergens; as well as non-nut plant allergens, which include wheat allergen; The composition comprises 10 or more different allergens, and each of the 10 or more different allergens has the same weight portion, wherein the composition is an oral preparation.

2. The mixed allergen composition according to claim 1, wherein the composition is a powder.

3. The mixed allergen composition according to claim 1, wherein the composition is a food supplement.

4. The mixed allergen composition according to claim 1, wherein the composition further comprises a biological agent, wherein the biological agent is a probiotic or a prebiotic.

5. The mixed allergen composition according to claim 1, wherein the non-nut plant allergen further comprises sesame allergen or oat allergen.

6. The mixed allergen composition according to claim 1, wherein the animal allergen further comprises crab allergen.

7. The mixed allergen composition according to claim 1, wherein the mixed allergen composition further comprises a soy allergen.

8. The mixed allergen composition of claim 1, wherein the peanut allergen is a peanut full allergen, the walnut allergen is a walnut full allergen, the hazelnut allergen is a hazelnut full allergen, the shrimp allergen is a shrimp full allergen, the egg allergen is an egg white full allergen, the milk allergen is a milk full allergen, the salmon allergen is a salmon full allergen, and the wheat allergen is a wheat full allergen.

9. The mixed allergen composition according to claim 5, wherein the sesame allergen is a sesame seed whole allergen, and the oat allergen is an oat whole allergen.

10. The mixed allergen composition according to claim 7, wherein the soy allergen is a soy whole allergen.

11. The mixed allergen composition according to claim 1, wherein the composition comprises 30 mg of each allergen present by weight of protein.

12. Use of the mixed allergen composition according to any one of claims 1 to 11 in the preparation of a medicament for treating allergy in a subject.

13. The use according to claim 12, wherein the subject is a teenager.

14. The use according to claim 12, wherein the subject is an adult.

15. The use according to claim 12, wherein the allergy is a food allergy.

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