Use of probiotics to modulate allergic immune responses

Abstract

The present invention relates to an effective amount of bacterial strains of the genera Lactobacillus and Lacticaseibacillus for the modulation of allergic immune responses in a subject, as well as methods and uses of said compositions.

Description

[Technical field]

[0001] The present invention relates to a novel use of a composition comprising a probiotic strain selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for the therapeutic modulation of an allergic immune response in a subject. The present invention further relates to a method for modulating an allergic immune response in a subject in need thereof. [Background technology]

[0002] Allergic diseases are estimated to affect 10-30% of people worldwide. For example, allergic rhinitis (AR) is a common disease that affects 500 million people worldwide. Although not life-threatening, AR severely impacts the quality of life and work efficiency of affected individuals and represents a significant economic burden to governments. Other diseases characterized by allergic immune responses include atopic eczema, allergic asthma, food allergies, as well as contact dermatitis (e.g., latex), drug anaphylaxis, and insect bite anaphylaxis.

[0003] An allergic reaction is an immunoglobulin (Ig) E-mediated allergic inflammatory reaction of mucosal or epithelial surfaces in which immune cells react to antigens, resulting in symptoms such as nasal / skin itching, sneezing, watery eye discharge, coughing and redness in the eyes.

[0004] Allergic reactions are characterized by the perturbation of T-helper type 1 (Th1) / T-helper type 2 (Th2) responses to allergens, specifically inducing an overly strong Th2 cytokine response, releasing interleukin (IL)-4, IL-5, IL-9, and IL-13, inducing IgE antibody formation, promoting the development and recruitment of eosinophils, and increasing mucus production in the airways, thereby causing allergic inflammation (Isolauri et al., 2001; Garn & Renz, 2007). Serum T-helper type 17 (Th17) cytokines, such as IL-17 and IL-22, are also increased, for example, in patients with allergic rhinitis (AR) and are associated with clinical symptoms, allergic inflammation, peripheral eosinophil counts, and the need for drug therapy (Ciprandi et al. 2009; Jordakieva and Jensen-Jarolim 2018).

[0005] Circulating Th2 and Th17 cytokine levels are reduced with allergen-specific immunotherapy, the only causative therapy currently available that alleviates symptoms associated with allergy or AR (Jordakieva and Jensen-Jarolim 2018).

[0006] Some probiotic bacteria have been shown to modulate immune responses and alleviate allergy symptoms, however, further evaluation of the effects and efficacy of probiotic bacteria is necessary to ensure the most potential effectiveness in treating allergies. Summary of the Invention [Problem to be solved by the invention]

[0007] The aim of the present invention was to evaluate the effects of the probiotic bacterial strains, Lactobacillus acidophilus La-14 (DGCC 11491) and Lacticaseibacillus paracasei Lpc-37 (DGCC 4981), on Th2 and Th17 cell responses of human peripheral blood mononuclear cells (PBMCs) stimulated with the T cell mitogen phytohemagglutinin (PHA). [Means for solving the problem]

[0008] In one aspect, the present invention relates to a therapeutic composition comprising an effective amount of one or more probiotic strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for use in modulating an allergic immune response in a subject.

[0009] In another aspect, the present invention relates to a non-therapeutic use of a composition for modulating an allergic immune response in a subject, said composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

[0010] In another aspect, the present invention relates to the use of a composition comprising an effective amount of one or more probiotic strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for the modulation of an allergic immune response in a subject.

[0011] In a further aspect, the present invention relates to a method of modulating an allergic immune response in a subject in need thereof, comprising administering to said subject a composition comprising an effective amount of one or more probiotic strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei. [Brief description of the drawings]

[0012] [Figure 1] Effect of La-14 on PHA-induced cytokine expression in PBMCs. Cells were treated with PHA alone or in combination with La-14. [Diagram 2] Effect of Lpc-37 on PHA-induced cytokine expression in PBMCs. Cells were treated with PHA alone or in combination with Lpc-37. [Diagram 3] Effect of La-14+Lpc-37 on PHA-induced cytokine expression in PBMCs. Cells were treated with PHA alone or in combination with La-14+Lpc-37. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Detailed aspects of the present invention are described below. Some detailed aspects are discussed in separate sections. This is for ease of reference and is in no way limiting. Unless the context clearly dictates otherwise, all embodiments described below apply equally to all aspects of the present invention.

[0014] bacteria The bacterial strain of the present invention is selected from the bacterial strains of the genus Lactobacillus and Lacticaseibacillus. Preferably, the bacterial strain of the present invention is of the species Lactobacillus acidophilus and Lacticaseibacillus paracasei. In particular, the bacterial strain is selected from the strain Lactobacillus acidophilus La-14 and the strain Lacticaseibacillus paracasei Lpc-37. Both the strains La-14 and Lpc-37 are commercially available from DuPont Nutrition Biosciences ApS.

[0015] The bacterial strains have also been deposited according to the Budapest Treaty by DuPont Nutrition Biosciences ApS, Langebrogade 1, DK-1411 Copenhagen K, Denmark, at the Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ), Inhoffenstrasse 7B, 38124 Braunschweig, Germany, and are registered under the following accession numbers: 1.La-14 strain (DGCC11491); deposited on June 1, 2021 under registration number DSM33880. 2. Strain Lpc-37 (DGCC4981; deposited on October 5, 2017 under accession number DSM32661).

[0016] Preferably, the bacterial strains used in the present invention are bacterial strains that are generally recognized as safe, and preferably have GRAS status, which is an American Food and Drug Administration (FDA) designation that a chemical or substance added to food is recognized by experts as safe and is therefore exempt from normal Federal Food, Drug, and Cosmetic Act (FFDCA) food additive tolerance requirements.

[0017] composition Although it is possible to administer the strains of the genus Lactobacillus and Lacticaseibacillus according to the present invention alone (i.e. without any support, diluent or excipient), said strains are typically administered on or in a support as part of a product, in particular as a component or at least one of the components of a composition, food ingredient, dietary supplement or pharma- ceutically acceptable composition or formulation.These products typically contain additional components that are well known to those skilled in the art.

[0018] Thus, in one embodiment, the present invention relates to a therapeutic composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for use in modulating an allergic immune response in a subject.

[0019] In another aspect, the present invention relates to a non-therapeutic use of a composition for modulating an allergic immune response in a subject, said composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

[0020] In a further embodiment, the present invention relates to a method of modulating an allergic immune response in a subject in need thereof, comprising administering to said subject a composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

[0021] It is understood that by modulating the allergic immune response, the bacterial strain reduces the production of Th1 and / or Th17 cytokines from human immune cells, thus improving the Th1 / Th2 balance and, consequently, reducing inflammation and allergic symptoms in allergic individuals.

[0022] The term "subject," as used herein, means a mammal.

[0023] In certain aspects of the invention, the subject can be a human.

[0024] In one embodiment, the subject may be female.

[0025] In one embodiment, the subject may be male.

[0026] In one embodiment, the subject can be of non-binary gender.

[0027] In one embodiment, the subject is not a child. As used herein, the term "child" refers to a human under the age of 7.

[0028] In one embodiment, the subject is a human aged 8 years or older.

[0029] In one embodiment, the subject is a human aged 16 years or older.

[0030] In one embodiment, the subject is a human aged 18 years or older.

[0031] In one embodiment, the subject is a healthy subject.

[0032] In a particular embodiment of the invention, the composition comprises bacterial strains from the species Lactobacillus acidophilus and from the species Lacticaseibacillus paracasei.

[0033] In one particular embodiment, the strain of the species Lactobacillus acidophilus is strain La-14. In another particular embodiment of the invention, the strain of the species Lacticaseibacillus paracasei is strain Lpc-37.

[0034] In a particular embodiment according to the invention, the bacterial strains of the species Lactobacillus acidophilus:Lacticaseibacillus paracasei are present in a ratio of 1:1 (v / v), preferably in a ratio of 1:2, more preferably in a ratio of 1:3, more preferably in a ratio of 1:4, more preferably in a ratio of 2:1, more preferably in a ratio of 3:1, more preferably in a ratio of 4:1.

[0035] In another particular embodiment, the bacteria present in the composition according to the invention are live bacteria.

[0036] In certain embodiments of the invention, modulation of the allergic immune response alleviates the symptoms of an allergic disease.

[0037] In another embodiment, the allergic disease is allergic rhinitis, hi another embodiment, the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis, and insect bite anaphylaxis.

[0038] In a further embodiment, the composition according to the present invention is encapsulated.

[0039] The bacterial strain according to the invention reduces the production of Th2 and immune cell derived cytokines, such as IL-5 and IL-13.

[0040] In a particular embodiment, the bacterial strain of the invention reduces the production of Th17 and immune cell-derived cytokines, such as IL-17 and IL-22.

[0041] The composition according to the invention may be present in different forms, such as a food, a food ingredient, a dietary supplement, or a pharma- ceutically acceptable composition or formulation.

[0042] In certain embodiments, the treatment regimen comprises oral administration of the composition.

[0043] In another specific embodiment, the treatment regimen comprises administering the composition for at least 1 week. In another embodiment, the treatment regimen comprises administering the composition for at least 2 weeks. In another embodiment, the treatment regimen comprises administering the composition at least twice a day.

[0044] Dosage The bacterial strains used according to the invention are at least 10 6 ~10 14 CFU of bacteria, more specifically 10 per gram of support 8 ~1012 CFU of bacteria, preferably 10 per gram of support 9 ~10 12 It may be present in CFU.

[0045] By "substrate" is meant a composition, food, food ingredient, dietary supplement, or pharma- ceutically acceptable composition.

[0046] Preferably, the bacterial strains of the genus Lacticaseibacillus and Lactobacillus used according to the present invention are present in an amount of about 10 per serving. 6 ~about 10 14 CFU of microorganisms, preferably about 10 per serving 8 ~about 10 12 CFU of microorganisms, more preferably about 10 per serving 9 ~about 10 11 Microorganisms may be administered in dosages of CFU, where CFU stands for "colony forming units."

[0047] The term "per serving" means that this amount of microorganism is given to a subject per day or per intake, preferably per day. For example, if the microorganism is administered in a food product (e.g., in yogurt), the yogurt preferably contains about 10 8 ~about 10 12 However, alternatively, this amount of microorganisms may be greater than or equal to about 10 CFU of microorganisms in total received by a subject during any particular time period, e.g., each 24 hour period. 6 ~about 10 12 CFU of microorganisms, preferably about 10 8 ~about 10 12 CFU of microorganisms, more preferably about 10 9 ~about 10 11 As long as the microorganisms are CFUs, they can be divided into five multiple doses, each with a smaller microorganism load.

[0048] In accordance with the present invention, the bacterial strain is at least 1×10 per dose. 6Colony forming units (CFU), preferably 1 x 10 6 ~1×10 14 , more preferably 1×10 8 ~1×10 12 The amount of strain per serving may refer to only one strain or may refer to two or more strains together.

[0049] food In one embodiment, the bacterial strain is used according to the invention in food products, such as milk-based food supplements, beverages or powders. As used herein, the term "food" is used in a broad sense to include human food and animal food (i.e., feed). In a preferred embodiment, the food product is for human consumption.

[0050] The food product may be in the form of a solution or in the form of a solid, depending on the purpose and / or the method of use and / or the method of administration.

[0051] When used as or in the preparation of a food, such as a functional food, the bacteria of the present invention may be used in combination with one or more of the following: a nutritionally acceptable carrier, a nutritionally acceptable diluent, a nutritionally acceptable excipient, a nutritionally acceptable adjuvant, a nutritionally active ingredient.

[0052] By way of example, the bacteria of the present invention can be used as an ingredient in soft drinks, fruit juices or beverages containing whey protein, health teas, cocoa drinks, milk drinks and lactobacillus drinks, yogurt and drinking yogurt, cheese, ice cream, frozen confections and desserts, confectionery, biscuit cakes and cake mixes, snack foods, nutritional foods and beverages, fruit fillings, protective glazes, chocolate bakery fillings, cheesecake flavoured fillings, fruit flavoured cake fillings, frostings for cakes and donuts, instant bakery filling creams, fillings for cookies, ready-to-eat bakery fillings, reduced calorie fillings, nutritional beverages for adults, plant-based milks, acidified soy / juice beverages, aseptic / retort chocolate beverages, bar mixes, beverage powders, calcium fortified soy / plain and chocolate milk, calcium fortified coffee beverages.

[0053] Advantageously, if the product is a food product, the bacterial strain should remain effective until the normal "sell by" or "best before" date when the food product is offered for sale by retailers. Preferably, the expiration date should extend beyond such date to the end of the normal freshness period when spoilage of the food product becomes evident. The desired length of time and normal shelf life will vary from food ingredient to food ingredient, and one skilled in the art will recognize that shelf life periods will vary with food ingredient type, food ingredient size, storage temperature, processing conditions, packaging materials, and packaging equipment age.

[0054] food ingredients The compositions of the present invention may take the form of a food and / or feed ingredient.

[0055] The term "food ingredient" or "feed ingredient" as used herein includes compositions that are, or can be added to, functional foods or foodstuffs as nutritional and / or dietary supplements for humans and animals.

[0056] The food ingredient may be in the form of a liquid, a suspension or a solid depending on the use and / or the method of application and / or administration.

[0057] Nutritional Supplements The compositions of the present invention may take the form of a dietary supplement or may be used in combination with dietary supplements, which themselves are also referred to herein as food supplements.

[0058] The term "dietary supplement" as used herein refers to a product intended for ingestion that contains "dietary ingredients" that are intended to add nutritional value or health benefits to the diet. "Dietary ingredients" may include, but are not limited to, one or any combination of the following substances: bacteria, probiotics (e.g., probiotic bacteria), vitamins, minerals, herbs or other botanicals, amino acids, dietary substances used by people to supplement their diet by increasing their total dietary intake, concentrates, metabolites, components, or extracts.

[0059] Dietary supplements can be found in many forms, such as tablets, capsules, soft gels, gel caps, liquids, or powders. Some dietary supplements help ensure adequate intake of essential nutrients from the diet, while others may help prevent or treat disease.

[0060] Medical Food The compositions of the present invention may take the form of a medical food.

[0061] "Medical food" means a food formulated to be taken or administered with or without medical supervision and intended for a specific dietary control or condition, with unique nutritional requirements established by medical evaluation based on recognized scientific principles.

[0062] Pharmaceutical Compositions The bacteria of the present invention can be used as or in the preparation of a pharmaceutical composition or formulation. As used herein, the term "pharmaceutical" is used in a broad sense to include human medicines and animal (i.e., veterinary) medicines.

[0063] In a preferred embodiment, the pharma- ceutically acceptable composition is a medicament.

[0064] The pharmaceutical compositions may be of therapeutic nature and may be curative, symptomatic or prophylactic in nature.

[0065] In a preferred embodiment of the invention, the medicament is for oral administration.

[0066] The pharma- ceutically acceptable compositions or supports may be, for example, in the form of creams, foams, gels, lotions, and ointments, compressed tablets, tablets, capsules, ointments, suppositories, sprays, such as nasal sprays, or drinkable solutions.

[0067] When used as a pharmaceutical product or in the preparation of a pharmaceutical product, the compositions of the present invention may be used in combination with one or more of the following: a pharma- ceutically acceptable carrier, a pharma- ceutically acceptable diluent, a pharma- ceutically acceptable excipient, a pharma- ceutically acceptable adjuvant, a pharma- ceutically active ingredient.

[0068] The pharmaceutical agent may be in the form of a solution or in the form of a solid, depending on the application and / or the method of use and / or the method of administration.

[0069] The bacterial strain of the present invention can be used as a pharmaceutical ingredient. As used herein, the composition can be the sole active ingredient or it can be at least one of a plurality (i.e., two or more) of active ingredients.

[0070] The bacterial strains according to the invention may be used in any suitable form, whether present alone or in combination with other components or ingredients. Similarly, combinations comprising the bacteria of the invention and other components and / or ingredients (i.e., ingredients such as food ingredients, functional food ingredients or pharmaceutical ingredients) may be used in any suitable dosage form.

[0071] The bacterial strain according to the present invention may be used in the form of solid or liquid formulations or alternative forms. Examples of solid formulations include, but are not limited to, tablets, capsules, powders, granules and powders, which may be water-dispersible, spray-dried or freeze-dried. Examples of liquid formulations include, but are not limited to, aqueous solutions, organic solutions or aqueous-organic solutions, suspensions and emulsions.

[0072] Suitable examples of dosage forms include one or more of the following: tablets, pills, capsules, ovoids, solutions or suspensions, optionally containing flavors or colorants, for immediate, delayed, modified, sustained, pulsed or controlled release applications.

[0073] By way of example, when the bacteria of the invention are used in tablet form, e.g. as a functional ingredient, the tablet may also contain one or more of the following: excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate and glycine; disintegrants such as starch (preferably corn starch, potato starch or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates; granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and gum arabic; lubricants such as magnesium stearate, stearic acid, glyceryl behenate and talc.

[0074] Examples of nutritionally acceptable carriers for use in preparing dosage forms include, for example, water, saline, alcohol, silicones, waxes, petrolatum, vegetable oils, polyethylene glycol, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, flavor oils, fatty acid 30 mono- and diglycerides, petroethrai fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and the like.

[0075] Preferred excipients for dosage forms include lactose, starch, a cellulose, milk sugar or high molecular weight polyethylene glycols.

[0076] For aqueous suspensions and / or elixirs, the bacteria of the present invention may be combined with various sweetening or flavoring agents, pigments or dyes, emulsifying and / or suspending agents, and diluents such as water, propylene glycol, and glycerin, as well as combinations thereof.

[0077] These dosage forms can also include gelatin capsules, dietary fiber capsules, dietary fiber tablets, and the like, or even dietary fiber drinks.

[0078] In one aspect, the bacteria according to the invention may be administered in the form of an aerosol, for example by nasal spray, for administration to the respiratory tract.

[0079] Prebiotics In one embodiment, the bacterial strains and compositions of the present invention may further be combined with or include one or more fibers and / or prebiotics.

[0080] Prebiotics are defined as substrates that are selectively utilized by the host microorganisms to confer health benefits. They are generally ingredients that beneficially affect the host's health by selectively stimulating the growth and / or activity of one or a limited number of types of bacteria, thus promoting the health of the host. Prebiotics can be applied for oral administration. Typically, prebiotics are carbohydrates (such as oligosaccharides), but the definition does not exclude non-carbohydrates such as polyphenols, or polyunsaturated fatty acids, or other ingredients that can be selectively utilized by a limited number of types of bacteria to confer health benefits. The most widespread forms of prebiotics are nutritionally classified as soluble fibers. Many forms of dietary fiber exhibit some level of prebiotic effect to some degree.

[0081] In one embodiment, prebiotics are selectively fermented ingredients that effect specific changes in both the composition and / or activity of the gastrointestinal or skin microflora that are beneficial to the well-being and health of the host.

[0082] Suitably, prebiotics may be used in accordance with the present invention in an amount of 0.01-100 g / day, preferably 0.1-50 g / day, more preferably 0.5-20 g / day. In one embodiment, prebiotics may be used in accordance with the present invention in an amount of 1-10 g / day, preferably 2-9 g / day, more preferably 3-8 g / day. In another embodiment, prebiotics may be used in accordance with the present invention in an amount of 5-50 g / day, preferably 5-25 g / day.

[0083] Examples of dietary sources of prebiotics include soybeans, inulin sources (such as Jerusalem artichoke, jicama and chicory root), raw oats, unrefined wheat and unrefined barley.

[0084] Examples of suitable prebiotics include alginates, xanthan, pectin, locust bean gum (LBG), inulin, guar gum, galactooligosaccharides (GOS), fructooligosaccharides (FOS), polydextrose 10 (i.e., Litesse®), lactitol, L-arabinose, D-xylose, L-rhamnose, D-mannose, L-fucose, inositol, sorbitol, mannitol, xylitol, fructose, carrageenan, alginates, microcrystalline cellulose (MCC), Betaine, lactosucrose, soybean oligosaccharides, isomaltulose (Palatinose™), isomaltooligosaccharides, glucooligosaccharides, xylooligosaccharides, mannooligosaccharides, beta-glucan, cellobiose, raffinose, gentiobiose, melibiose, xylobiose, cyciodextrin, isomaltose, trehalose, stachyose, panose, pullulan, verbascose, galactomannan, (human) milk oligosaccharides, and all forms of resistant starch.

[0085] The combination of one or more of the bacterial strains according to the invention with one or more fibres and / or prebiotics according to the invention exhibits synergistic effects in certain applications (i.e. effects that are greater than the additive effects of the bacteria when used separately).

[0086] In one embodiment, the bacterial strain or mixtures thereof according to the invention are used in combination with one or more fibres and / or prebiotics.

[0087] Preferably, the prebiotics used are polydextrose, lactitol, inositol, L-arabinose, D-xylose, L-rhamnose, D-mannose, L-fucose, sorbitol, mannitol, xylitol, fructose, carrageenan, alginate, 5 microcrystalline cellulose (MCC), milk oligosaccharides or betaine.

[0088] In a further aspect, the present invention relates to a composition, a food, a food ingredient, a dietary supplement or a pharma- ceutically acceptable composition comprising a bacterial strain according to the invention or a mixture thereof and one or more fibres and / or prebiotics.

[0089] EMBODIMENTS OF THE PRESENTINVENTION For the avoidance of doubt, some embodiments to which the present invention pertains are described below.

[0090] Embodiment 1. A therapeutic composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for use in modulating an allergic immune response in a subject.

[0091] Embodiment 2. Non-therapeutic use of a composition for modulating an allergic immune response in a subject, said composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

[0092] Embodiment 3. The composition of embodiment 1 or 2, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and from the species Lacticaseibacillus paracasei.

[0093] Embodiment 4. A composition according to any one of embodiments 1 to 3, wherein the bacterial strain of the species Lactobacillus acidophilus is strain La-14.

[0094] Embodiment 5. The composition according to any one of embodiments 1 to 3, wherein the bacterial strain of the species Lacticaseibacillus paracasei is the Lpc-37 strain.

[0095] Embodiment 6. The bacterial strain is at least 1 x 10 per dose. 6 Colony forming units (CFU), preferably 1 x 10 6 ~1×10 14 , more preferably 1×10 8 ~1×10 12 6. The composition of any one of embodiments 1 to 5, wherein

[0096] Embodiment 7. The composition of any one of embodiments 1 to 6, wherein said modulation of an allergic immune response alleviates a symptom of an allergic disease.

[0097] Embodiment 8 The composition of embodiment 7, wherein the allergic disease is allergic rhinitis.

[0098] Embodiment 9. The composition according to embodiment 7, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

[0099] Embodiment 10. The composition of any one of embodiments 1 to 6, wherein the bacterial strain reduces production of Th2 and immune cell-derived cytokines.

[0100] Embodiment 11. The composition of any one of embodiments 1 to 6, wherein the bacterial strain reduces production of Th17 and immune cell-derived cytokines.

[0101] Embodiment 12. The composition of embodiment 10, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

[0102] Embodiment 13 The composition of embodiment 11, wherein the Th17 and immune cell-derived cytokines are IL-17 and IL-22.

[0103] Embodiment 14. The composition of any one of embodiments 1 to 13, wherein the composition is a food product, a food ingredient, a dietary supplement, or a pharma- ceutically acceptable composition.

[0104] Embodiment 15. The composition of embodiment 14, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray.

[0105] Embodiment 16. Use of a composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for modulating an allergic immune response in a subject.

[0106] Embodiment 17. The use according to embodiment 16, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and from the species Lacticaseibacillus paracasei.

[0107] Embodiment 18. Use according to embodiment 16 or 17, wherein the bacterial strain of the species Lactobacillus acidophilus is strain La-14.

[0108] Embodiment 19. The use according to embodiment 16 or 17, wherein the bacterial strain of the species Lacticaseibacillus paracasei is the Lpc-37 strain.

[0109] Embodiment 20. The bacterial strain comprises at least 1 x 10 6 Colony forming units (CFU), preferably 1 x 10 6 ~1×1014 , more preferably 1×10 8 ~1×10 12 20. The use according to any one of embodiments 16 to 19, wherein

[0110] Embodiment 21. The use according to any one of embodiments 16 to 20, wherein said modulation of an allergic immune response alleviates a symptom of an allergic disease.

[0111] Embodiment 22. The use according to embodiment 21, wherein the allergic disease is allergic rhinitis.

[0112] Embodiment 23. The use according to embodiment 21, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

[0113] Embodiment 24. The use of any one of embodiments 16 to 20, wherein the bacterial strain reduces the production of Th2 and immune cell-derived cytokines.

[0114] Embodiment 25. The use according to any one of embodiments 16 to 20, wherein the bacterial strain reduces the production of Th17 and immune cell-derived cytokines.

[0115] Embodiment 26 The use according to embodiment 24, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

[0116] Embodiment 27 The use according to embodiment 25, wherein the Th17 and immune cell derived cytokines are IL-17 and IL-22.

[0117] Embodiment 28. The use of any one of embodiments 16 to 27, wherein the composition is a food product, a food ingredient, a dietary supplement or a pharma- ceutically acceptable composition.

[0118] Embodiment 29. The use according to embodiment 28, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray.

[0119] Embodiment 30. A method for modulating an allergic immune response in a subject in need thereof, comprising administering to the subject a composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

[0120] Embodiment 31. The method of embodiment 30, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and Lacticaseibacillus paracasei.

[0121] Embodiment 32. The method according to embodiment 30 or 31, wherein the bacterial strain of the species Lactobacillus acidophilus is strain La-14.

[0122] Embodiment 33. The method according to claim 30 or 31, wherein the bacterial strain of the species Lacticaseibacillus paracasei is the Lpc-37 strain.

[0123] Embodiment 34. The bacterial strain has a concentration of at least 1 × 10 per dose. 6 Colony forming units (CFU), preferably 1 x 10 6 ~1×10 14 , more preferably 1×10 8 ~1×10 12 34. The method according to any one of embodiments 30 to 33, wherein

[0124] Embodiment 35. The method of any one of embodiments 30-34, wherein said modulation of an allergic immune response alleviates a symptom of an allergic disease.

[0125] Embodiment 36 The method of embodiment 35, wherein the allergic disease is allergic rhinitis.

[0126] Embodiment 37. The method of embodiment 36, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

[0127] Embodiment 38. The composition of any one of embodiments 30 to 34, wherein the bacterial strain reduces production of Th2 and immune cell-derived cytokines.

[0128] Embodiment 39. The method of any one of embodiments 30 to 34, wherein the bacterial strain reduces production of Th17 and immune cell-derived cytokines.

[0129] Embodiment 40 The composition of embodiment 38, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

[0130] Embodiment 41 The composition of embodiment 39, wherein the Th17 and immune cell-derived cytokines are IL-17 and IL-22.

[0131] Embodiment 42. The method of any one of embodiments 30 to 41, wherein the composition is a food product, a food ingredient, a dietary supplement, or a pharma- ceutically acceptable composition.

[0132] Embodiment 43 The method of embodiment 42, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray. EXAMPLES

[0133] The following examples are provided to demonstrate and further illustrate specific embodiments and aspects of the present invention and should not be construed as limiting the scope of the invention.

[0134] Materials and Methods Strains and growth conditions The effects of Lactobacillus acidophilus La-14 (DGCC 11491) and Lacticaseibacillus paracasei Lpc-37 (DGCC 4981) on Th2 and Th17 cell responses in human peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin (PHA), a common ex vivo model for evaluating allergic responses, were evaluated.

[0135] PBMCs were isolated from buffy coats purchased from the Finnish Red Cross Blood Service, following the permission of the Helsinki and Uusimaa Hospital District ethical committee (approval: HUS / 828 / 2020). Buffy coats were diluted 1:1 with 2 mM EDTA (Gibco) in phosphate-buffered saline (PBS, Thermo Fisher Scientific), and PBMCs were isolated by Ficoll-Paque (GE Healthcare) density gradient centrifugation in SepMate tubes (Stemcell technologies, Grenoble, France). After washing the cells with PBS (Thermo Fisher Scientific), the cells were counted and diluted in cell culture medium RPMI1640 (Merck) supplemented with 10% fetal bovine serum and 1% antibiotic-antimycotic (both Thermo Fisher Scientific). Cells were plated into non-tissue-treated 24-well plates (Falcon) at 10^6 cells / well.

[0136] Prebiotic bacterial strains were used in the experiments after overnight subculture. Probiotics were harvested by centrifugation, washed once with PBS (Thermo Fisher Scientific), and suspended in cell culture medium RPMI1640. OD600 was adjusted to correspond to a given bacteria:host cell ratio of 10:1. Bacterial counts were calculated by flow cytometry prior to the experiments, and a trend line was established for the growth curve OD values ​​of the strains. This trend line allowed the calculation of bacterial counts based on OD values ​​without the need to calculate the amount of bacteria individually for each experiment.

[0137] Phytohemagglutinin L (PHA-L, Merck) ("PHA") was dissolved in RPMI1640 (serum- and antibiotic-free) and stored in aliquots at -20°C. PHA-L was used at a concentration of 1 μg / ml. PBMCs were treated with individual probiotic strains or a combination of the two strains with PHA challenge in triplicate wells. As a baseline control, PBMCs were treated with PHA challenge without probiotics. Supernatant samples for ELISA testing were taken after 24 h incubation and stored at -80°C until analysis.

[0138] Cell culture supernatants of PBMC cell cultures were analyzed for IL-5, IL-13, IL-17A, and IL-22 cytokines using human ELISA kits (Quanterix, Billerica, MA, USA). Results were analyzed with Quanterix SP-X analysis software and graphed in box plots (Figures 1, 2, and 3). Box plots were generated for the average values ​​of triplicate wells of four PBMC donors normalized to PHA control samples.

[0139] statistical analysis The mean values ​​of three replicate wells were used for analysis. The primary efficacy variables of the study were the percent change relative to the PHA control for each cytokine, calculated for each probiotic treatment as the measured cytokine value divided by the mean of the three PHA control values ​​from the same donor. Differences relative to the PHA control were assessed by using a repeated measures analysis of variance (RM-ANOVA) with probiotic as the main effect and donor as a random effect in the model. Probiotic estimates were compared to 1 (not significantly different) and fold change comparisons with the PHA control were performed in the same analysis model using contrasts. P values ​​and 95% confidence intervals were estimated for all comparisons.

[0140] result The effect of the La-14 strain on PHA-induced cytokine expression in PBMCs was examined. Cells were treated with PHA alone or in combination with La-14. Cytokine concentrations were measured from the supernatants, and the results show that the La-14 strain induced IL-5, IL-13, and IL-17 responses were statistically significantly reduced compared to PHA stimulation alone (Figure 1).

[0141] The effect of the Lpc-37 strain on PHA-induced cytokine expression in PBMC was examined. Cells were treated with PHA alone or in combination with Lpc-37. Cytokine concentrations were measured from the supernatants, and the results show that the IL-5 and IL-13 responses induced by the Lpc-37 strain were statistically significantly reduced compared to PHA stimulation alone (Figure 2).

[0142] The effect of the combination of La-14 and Lpc-37 strains on PHA-induced cytokine expression in PBMCs was examined. Cells were treated with PHA alone or in combination with the combination of La-14 and Lpc-37 strains. Cytokine concentrations were measured from the supernatants, and the results show that the IL-5, IL-13, IL-17, and IL-22 responses with the combination of La-14 and Lpc-37 strains are statistically significantly reduced compared to PHA stimulation alone (Figure 3).

[0143] The present invention shows that Lactobacillus acidophilus La-14, Lacticaseibacillus paracasei Lpc-37, and their combinations reduce the expression of Th2 and Th17 cytokines (IL-5, IL-13, IL-17, and IL-22) in PHA-stimulated PBMCs (Figure 3). Although the expression of IL-17 is not reduced by Lpc-37 alone, and the expression of IL-22 cytokines is not reduced by La-14 or Lpc-37 alone (Figures 1 and 2), the combination of La-14 and Lpc-37 can significantly reduce IL-17 and IL-22, indicating the synergistic effect of these strains (Figure 3).

[0144] The results of the present invention show that probiotics La-14 and Lpc-37 can downregulate allergen-specific immune responses and prevent or alleviate the symptoms of allergic diseases such as allergic rhinitis, food allergies, atopic dermatitis, and allergic asthma.

[0145] All publications mentioned herein above are incorporated herein by reference. Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the art of biochemistry and biotechnology or related fields are intended to be within the scope of the following claims.

[0146] References Ciprandi G,De Amici M,Murdaca G,Fenoglio D,Ricciardolo F,Marseglia G,Tosca M.(2009)Serum interleukin-17 levels are related to clinical severity in allergic rhinitis.Allergy.64:1375-1378.https: / / doi.org / 10.1111 / j.1398-9995.2009.02010.x Garn H,Renz H.(2007)Epidemiological and immunological evidence for the hygiene hypothesis.Immunobiology.212:441-52.doi:10.1016 / j.imbio.2007.03.006.Epub 2007 Apr 30.PMID:17544829。 Isolauri E,Suetas Y,Kankaanpaeae P,Arvilommi H,Salminen S.(2001)Probiotics:effects on immunity.Am J Clin Nutr.73:444S-450S.doi:10.1093 / ajcn / 73.2.444s.PMID:11157355。 Jordakieva G,Jensen-Jarolim E.(2018)The impact of allergen exposure and specific immunotherapy on circulating blood cells in allergic rhinitis.World Allergy Organ J.11:19.doi:10.1186 / s40413-018-0197-0.PMID:30128065;PMCID:PMC6092783。

Claims

1. A therapeutic composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for use in modulating an allergic immune response in a subject.

2. 1. A non-therapeutic use of a composition for modulating an allergic immune response in a subject, said composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

3. 3. The composition according to claim 1 or 2, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and from the species Lacticaseibacillus paracasei.

4. The composition according to any one of claims 1 to 3, wherein said bacterial strain of the species Lactobacillus acidophilus is strain La-14.

5. The composition according to any one of claims 1 to 3, wherein the bacterial strain of the species Lacticaseibacillus paracasei is strain Lpc-37.

6. The bacterial strain is at least 1 x 10 per dose ６ Colony forming units (CFU), preferably 1 x 10 ６ ~1×10 １４ , more preferably 1×10 ８ ~1×10 １２ The composition of any one of claims 1 to 5, wherein

7. The composition of any one of claims 1 to 6, wherein said modulation of the allergic immune response alleviates symptoms of an allergic disease.

8. The composition according to claim 7, wherein the allergic disease is allergic rhinitis.

9. The composition according to claim 7, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

10. The composition of any one of claims 1 to 6, wherein the bacterial strain reduces production of Th2 and immune cell-derived cytokines.

11. The composition of any one of claims 1 to 6, wherein the bacterial strain reduces the production of Th17 and immune cell-derived cytokines.

12. The composition of claim 10, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

13. The composition of claim 11, wherein the Th17 and immune cell-derived cytokines are IL-17 and IL-22.

14. The composition according to any one of claims 1 to 13, wherein the composition is a food product, a food ingredient, a dietary supplement or a pharma- ceutically acceptable composition.

15. The composition of claim 14, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray.

16. 13. Use of a composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei for modulating an allergic immune response in a subject.

17. 17. The use according to claim 16, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and from the species Lacticaseibacillus paracasei.

18. The use according to claim 16 or 17, wherein the bacterial strain of the species Lactobacillus acidophilus is strain La-14.

19. 18. Use according to claim 16 or 17, wherein the bacterial strain of the species Lacticaseibacillus paracasei is strain Lpc-37.

20. The bacterial strain is at least 1 x 10 per dose ６ Colony forming units (CFU), preferably 1 x 10 ６ ~1×10 １４ , more preferably 1×10 ８ ~1×10 １２ The use according to any one of claims 16 to 19, wherein

21. The use according to any one of claims 16 to 20, wherein said modulation of the allergic immune response alleviates the symptoms of an allergic disease.

22. The use according to claim 21 , wherein the allergic disease is allergic rhinitis.

23. 22. The use according to claim 21, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

24. The use according to any one of claims 16 to 20, wherein the bacterial strain reduces the production of Th2 and immune cell derived cytokines.

25. The use according to any one of claims 16 to 20, wherein the bacterial strain reduces the production of Th17 and immune cell-derived cytokines.

26. The use according to claim 24, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

27. The use according to claim 25, wherein the Th17 and immune cell derived cytokines are IL-17 and IL-22.

28. The use according to any one of claims 16 to 27, wherein the composition is a food product, a food ingredient, a dietary supplement or a pharma- ceutically acceptable composition.

29. 29. The use according to claim 28, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray.

30. A method of modulating an allergic immune response in a subject in need thereof, comprising administering to the subject a composition comprising an effective amount of one or more bacterial strains selected from the species Lactobacillus acidophilus and / or Lacticaseibacillus paracasei.

31. 31. The method of claim 30, wherein the composition comprises bacterial strains from the species Lactobacillus acidophilus and Lacticaseibacillus paracasei.

32. 32. The method according to claim 30 or 31, wherein the bacterial strain of the species Lactobacillus acidophilus is strain La-14.

33. 32. The method according to claim 30 or 31, wherein the bacterial strain of the species Lacticaseibacillus paracasei is strain Lpc-37.

34. The bacterial strain is at least 1 x 10 per dose ６ Colony forming units (CFU), preferably 1 x 10 ６ ~1×10 １４ , more preferably 1×10 ８ ~1×10 １２ The method according to any one of claims 30 to 33, wherein

35. The method of any one of claims 30 to 34, wherein said modulation of an allergic immune response alleviates a symptom of an allergic disease.

36. 36. The method of claim 35, wherein the allergic disease is allergic rhinitis.

37. 37. The method of claim 36, wherein the allergic disease is atopic eczema, allergic asthma, food allergy, contact dermatitis, drug anaphylaxis and insect bite anaphylaxis.

38. The composition of any one of claims 30 to 34, wherein the bacterial strain reduces production of Th2 and immune cell-derived cytokines.

39. The method of any one of claims 30 to 34, wherein the bacterial strain reduces production of Th17 and immune cell-derived cytokines.

40. The composition of claim 38, wherein the Th2 and immune cell derived cytokines are IL-5 and IL-13.

41. The composition of claim 39, wherein the Th17 and immune cell-derived cytokines are IL-17 and IL-22.

42. The method of any one of claims 30 to 41, wherein the composition is a food product, a food ingredient, a dietary supplement or a pharma- ceutically acceptable composition.

43. 43. The method of claim 42, wherein the pharmaceutical composition is in the form of a cream or a spray, such as a nasal spray.

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