Use of lactiplantic Bacillus plantarum strains to improve allergic skin conditions

Abstract

This invention relates to the use of a Lactiplantibacillus plantarum strain to improve skin conditions caused by allergies, and in one embodiment, the strain and its derived disrupted solution, culture solution, extract of the culture solution, or mixtures thereof can be effectively used to prevent, improve, or treat allergic diseases.

Description

[Technical Field]

[0001] The present invention relates to the use of lactiplantic Bacillus plantarum strains to improve allergic skin conditions. [Background technology]

[0002] Allergy is a compound word derived from the Greek words "allos," meaning "to change," and "ergo," meaning "to act," and refers to an abnormal biological reaction. As can be inferred from the etymology, allergies refer to congenital or acquired immune abnormalities that cause hypersensitivity reactions, such as hives, itching, runny nose, and coughing, in certain individuals due to allergens, substances that normally do not cause any reaction in humans. Typical allergens include pollen, drugs, plant fibers, bacteria, fungi, dust mites, food (egg albumin, milk protein, peanuts), hair dye, and chemicals. Recently, these allergic diseases have been on the rise worldwide due to factors such as increased indoor living, an increase in allergens such as new materials and chemicals, environmental pollution, weakened immune function due to stress, and changes in diet. Representative allergic diseases include atopic dermatitis, bronchial asthma, allergic rhinitis, allergic keratitis, and urticaria, and mast cells are known to be important mediators of these allergic diseases.

[0003] Allergic reactions are induced by the activation of intracellular signaling pathways when allergens bind to IgE bound to the high-affinity IgE receptor (FcεRI) on the surface of mast cells. Specifically, when an allergen enters the body, T-helper cells and B cells are sequentially activated, resulting in the production of IgE. The allergen is recognized by antigen-presenting cells, which differentiate Th0 (T0 helper) cells into Th2 cells through allergen presentation. These differentiated Th2 cells secrete cytokines, such as IL-4, IL-5, and IL-13, which promote the development of eosinophils in the bone marrow and induce their introduction into inflamed tissues, as well as act on B cells to induce IgE production. The IgE thus produced binds to the high-affinity IgE receptor (FcRIα) on the mast cell membrane, and when allergens, such as dust mites and pollen, bind to this receptor and form cross-links, the receptor (FcRIα) aggregates. Mast cells are activated by the cross-linking and aggregation of receptors (FcRIα), resulting in an increase in intracytoplasmic calcium ion (Ca++) concentration. The increase in calcium ion concentration acts on various intracellular regulatory factors, such as cholesterol in the cell membrane and the cytoskeleton, to migrate granules to the cell membrane, leading to degranulation through granule vacuolation, granule fusion with each other, and granule-to-cell membrane fusion. Degranulation leads to the secretion of histamine, prostaglandins, leukotrienes, and serotonin, resulting in vasodilation, increased vascular permeability, and bronchial smooth muscle contraction. Mast cells secrete inflammatory cytokines such as IL-1β, IL-4, IL-6, and TNF-α, which induce the infiltration of neutrophils, eosinophils, macrophages, Th2 cells, and basophils into the affected tissue, inducing an allergic inflammatory response.

[0004] Therefore, substances that inhibit mast cell activation, degranulation, or the production of inflammatory cytokines could be effective therapeutic agents for allergies. Currently, anti-allergy drugs used include antihistamines (e.g., alkylamines, desloratadine), mast cell stabilizers (e.g., cromoglycate), leukotriene antagonists (e.g., montelukast), and anti-IgE antibodies (e.g., omalizumab). However, the effects of these drugs are often temporary, and sustained treatment with high concentrations can induce side effects, limiting their use in the treatment of chronic immune disorders such as allergic diseases.

[0005] Therefore, there is still a need for the development of new substances that have the effect of preventing and improving allergic diseases, while having fewer side effects and having a long-lasting effect. Summary of the Invention [Problem to be solved by the invention]

[0006] One aspect is to provide a Lactiplantibacillus plantarum strain having antiallergic activity, wherein the antiallergic activity is to reduce serum IgE levels or inhibit the degranulation activity of mast cells.

[0007] Another aspect provides the Lactiplantibacillus plantarum HD-02 strain belonging to the genus Lactiplantibacillus, deposited under accession number KACC81228BP.

[0008] Another aspect is to provide a lysate, culture medium, or culture medium extract from said strain.

[0009] Another aspect is to provide a food composition comprising a lactiplantibacillus plantarum strain, a lysate derived from said strain, a culture broth or a mixture thereof as an active ingredient.

[0010] Another aspect is to provide a health functional food for preventing or ameliorating allergic diseases, which contains a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0011] Another aspect is to provide a pharmaceutical composition for treating or preventing an allergic disease, comprising a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof, as an active ingredient.

[0012] Another aspect is to provide a cosmetic composition for preventing or ameliorating allergic diseases, which comprises a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0013] Another aspect is to provide a feed composition for preventing or ameliorating allergic diseases, which comprises a lactiplantibacillus plantarum strain, a lysate derived from said strain, a culture solution, or a mixture thereof as an active ingredient.

[0014] Another aspect provides a method for preventing or treating an allergic disease, comprising administering to an individual in need thereof an effective amount of the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a culture of said strain, a lysate of said strain, or a composition comprising a mixture thereof.

[0015] Another aspect provides use of a composition comprising the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof, for the manufacture of a pharmaceutical preparation or a functional health food preparation for preventing or treating an allergic disease. [Means for solving the problem]

[0016] One embodiment provides a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus.

[0017] Lactiplantibacillus is a gram-positive, aerobic or facultatively anaerobic bacillus that is widely distributed in nature. Microorganisms belonging to the genus Lactiplantibacillus include Lactiplantibacillus plantarum and Lactiplantibacillus fermentum. The present inventors conducted research to develop a new strain with excellent anti-allergy effects and selected Lactiplantibacillus Plantanum HD-02 as a candidate anti-allergy strain. This strain was deposited at the Agricultural Genetic Resources Center on August 26, 2022, under accession number KACC81228BP. This strain corresponds to a probiotic strain and is harmless to the human body, allowing it to be used without side effects.

[0018] In one embodiment, the Lactiplantibacillus plantarum strain may be the strain deposited under accession number KACC81228BP.

[0019] In one embodiment, the Lactiplantibacillus plantarum strain may be a strain comprising the 16S rRNA sequence of SEQ ID NO:1.

[0020] In one embodiment, the strain may have a 16S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1 or a 16S rRNA containing a nucleotide sequence having 97% or more identity thereto, specifically, at least 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% identity to the nucleotide sequence of SEQ ID NO: 1 herein.

[0021] In one embodiment, the Lactiplantibacillus plantarum strain may be a strain comprising the recA gene sequence of SEQ ID NO:2.

[0022] In one embodiment, the strain may have a recA gene sequence of SEQ ID NO: 2 or a gene sequence having 97% or more homology thereto, specifically at least 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% homology to the gene sequence of SEQ ID NO: 2 herein.

[0023] In one embodiment, the strain may be a naturally occurring mutant strain.

[0024] In one embodiment, the bacterial strain may be a live cell, a killed cell, or a cytoplasmic fraction obtained by disrupting the strain, preferably a live cell. The bacterial strain may be present as a live or killed cell, and may be present in a dried or lyophilized form.

[0025] In one embodiment, the strain may have anti-allergic (hereinafter, allergic and anti-allergic may be used interchangeably) activity.

[0026] In one embodiment, the strain may have the activity of suppressing Th2-mediated cytokine production, suppressing mast cell degranulation activity, suppressing serum IgE levels, or suppressing serum MCPT-1 levels. Specifically, the suppression of mast cell degranulation activity may be suppressing the secretion of β-hexosaminidase in activated mast cells.

[0027] In one embodiment, the strain may reduce serum IgE levels or reduce the binding of IgE to surface receptors on mast cells, and may also suppress serum MCPT-1 levels.

[0028] Therefore, the strain according to one embodiment can be useful in treating allergic diseases.

[0029] Another embodiment provides a lysate, culture broth, culture broth extract, or mixture thereof from a strain of Lactiplantibacillus plantarum.

[0030] The term "culture medium" as used herein can be used interchangeably with "culture," "culture supernatant," "culture supernatant," "conditioned culture medium," or "conditioned medium," and can refer to the entire medium containing the strain, its metabolites, and additional nutrients obtained by culturing a Lactipranchibacillus strain for a certain period of time in a medium that can provide nutrients so that the strain can grow and survive in vitro. The culture refers to the product obtained by culturing a probiotic strain in a known medium, and the product may or may not contain the strain itself. The medium can be selected from known liquid or solid media, such as, but not limited to, MRS liquid medium, GAM liquid medium, MRS agar medium, GAM agar medium, and BL agar medium.

[0031] The culture medium may include the culture medium itself obtained by culturing the strain, a concentrate thereof, or a freeze-dried product thereof, or a culture supernatant obtained by removing the strain from the culture medium, or a concentrate or freeze-dried product thereof.

[0032] The term "lysate" herein can be used interchangeably with "lysate" and refers to an aqueous medium solution or suspension of disrupted microbial cells, such as Lactiplantibacterium plantarum. Cell lysates contain macromolecules, such as DNA, RNA, proteins, peptides, carbohydrates, lipids, and / or micromolecules, such as amino acids, sugars, fatty acids, or fractions thereof. Additionally, the lysate contains cell debris, which may be smooth or granular in structure.

[0033] As used herein, the term "culture extract" refers to a substance extracted from the culture medium or a concentrate thereof, and may include an extract, a diluted or concentrated extract, a dried product obtained by drying the extract, or a preparation or purified product thereof, or a fraction thereof.

[0034] The culture of the present invention may include a viable culture (i.e., a product obtained by culturing viable bacteria in a medium), a culture in which viable bacteria have been killed in the viable culture, or a culture filtrate (i.e., a product obtained by removing viable or killed bacteria from the culture). Furthermore, components derived from the strains of the present application may include, for example, cell wall structures obtained by disrupting the cells, or cytoplasmic fractions containing extracellular vesicles (EVs). Furthermore, the forms of lactic acid bacteria suitable for inclusion in pharmaceutical compositions, formulation methods, and methods for isolating strain-derived components are well known to those skilled in the art.

[0035] In one embodiment, the culture medium may be obtained by culturing Lactiplantibacillus plantarum in an appropriate medium (e.g., MRS plate medium) at a temperature above 10°C or below 40°C for a certain period of time, for example, 4 to 50 hours.

[0036] In one embodiment, the culture medium and culture conditions for culturing the Lactiplantibacillus plantarum can be appropriately selected or modified by a person skilled in the art.

[0037] Another aspect provides a composition comprising a lysate, a culture broth, an extract of a culture broth, or a mixture thereof from a strain of Lactiplantibacillus plantarum.

[0038] In one embodiment, the composition may further comprise an additive selected from the group consisting of preservatives, dyes, emulsifiers, sweeteners, stabilizers, flavor enhancers, flavoring agents, and acidulants.

[0039] In one embodiment, the dosage form may be at least one selected from the group consisting of a solution, an emulsion, a viscous mixture, a powder, a granule, a tablet, and a capsule.

[0040] In one embodiment, the composition may have anti-allergic activity.

[0041] In one embodiment, the anti-allergic activity of the strain as an active ingredient can be suppressing the production of Th2 (helper T2 cell)-mediated cytokines or alleviating allergic reactions, which can include alleviating immune hypersensitivity reactions, specifically improving or alleviating skin conditions caused by immune hypersensitivity reactions, such as atopic dermatitis.

[0042] In one embodiment, the anti-allergic activity can include inhibiting degranulation in activated mast cells, specifically inhibiting the secretion of β-hexosaminidase, histamine, prostaglandins, and serine proteases in activated mast cells.

[0043] In one embodiment, the anti-allergic activity may include reducing serum IgE levels or reducing the binding of IgE to surface receptors on mast cells, and may further include suppressing serum MCPT-1 levels.

[0044] According to one embodiment, the anti-allergic activity may include the activity of inhibiting mast cell degranulation by 50% to 70% when the Lactipranchibacillus plantarum HD-02 strain is treated in mast cells RBL-2H3. According to one embodiment, the anti-allergic activity may include the activity of inhibiting allergic reactions by 20% to 40% when the Lactipranchibacillus plantarum HD-02 strain is treated in an animal model of anti-DNP IgE-induced passive cutaneous anaphylaxis (PCA).

[0045] According to one embodiment, the antiallergic activity may include the activity of suppressing ear edema by 40% to 60%, the activity of reducing serum IgE concentration by 50% to 70%, or the activity of reducing serum MCPT-1 concentration by 50% to 70% when the Lactiplantibacillus plantarum HD-02 strain is administered to an animal model of MC903-induced atopic dermatitis.

[0046] According to one embodiment, the antiallergic activity may include, when the Lactiplantibacillus plantarum HD-02 strain is administered to an animal model of house dust mite-induced atopic dermatitis (a severe atopic dermatitis model), the activity of reducing serum IgE levels by 50% to 70%, the activity of reducing transepidermal water loss by 30% to 60%, the activity of relieving itching by 40% to 70%, the activity of reducing the number of immune cells (specifically, B cells, T cells, Th1 cells, and Th2 cells) activated in lymph nodes (e.g., axillary lymph nodes) due to immune hypersensitivity reactions by 50% to 90%, the activity of suppressing edema by 40% to 60%, or the activity of reducing the number of activated mast cells in inflamed tissues by 50% to 70%.

[0047] In one embodiment, the Lactiplantibacillus plantarum HD-02 strain may have activity that reduces the symptoms of allergic reactions or atopic dermatitis to a level of 90% or less, 80% or less, 70% or less, 67% or less, 10-90%, 10-80%, 10-70%, 10-67%, 20-90%, 20-80%, 20-70%, 20-67%, 30-90%, 30-80%, 30-70%, 30-67%, 40-90%, 40-80%, 40-70%, 40-67%, 50-90%, 50-80%, 50-70%, 50-67%, or 60-67%, based on 100% of the symptoms of allergic reactions in a negative control group in which allergic reactions or atopic dermatitis are induced without administration of the Lactiplantibacillus plantarum HD-02 strain.

[0048] Another aspect provides for the disease-modifying, prophylactic or therapeutic use of Lactiplantibacillus plantarum, a lysate, a culture broth or an extract of a culture broth of said strain.

[0049] In one embodiment, the use may include the prevention, amelioration, or treatment of a disease or condition mediated by IgE (Immunoglobulin E). Specifically, the disease mediated by IgE (Immunoglobulin E) may be an allergic disease.

[0050] As used herein, the term "allergic disease" refers to hypersensitivity to a specific substance in the human body, i.e., a disease, illness, or abnormal state caused by an excessive immune system response to a substance introduced from the outside, where the substance introduced from the outside may be an allergen, which is an antigen that causes the allergic disease. The allergy may refer to a hypersensitivity reaction in which a disease is induced by the release of inflammatory mediators such as histamine due to an introduced substance from the outside, and the hypersensitivity reaction may be a type 1 hypersensitivity reaction, a type 2 hypersensitivity reaction, a type 3 hypersensitivity reaction, or a type 4 hypersensitivity reaction.

[0051] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0052] Non-limiting examples of the allergic disease include, but are not limited to, one or more diseases selected from the group consisting of edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic tympanitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, dry eye, pruritus, anaphylactic shock, hay fever, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0053] As used herein, the term "atopic dermatitis" refers to a chronically recurring, severely itchy skin disease in which the epidermis thickens and multiple immune response cells infiltrate. The skin of patients with atopic dermatitis is dry and lackluster. Xeroderma skin has a weakened skin barrier, leading to frequent moisture loss and allowing external irritants to easily penetrate the skin.

[0054] In the acute stage of atopic dermatitis, severely itchy red spots, small raised papules, blisters, scars, and pus appear, sometimes accompanied by secondary infection. As the lesions progress, scars and red papules covered with scales may appear in the subacute stage. In the chronic stage, repeated scratching can lead to lichenification, where the skin becomes rough and thick, and wrinkles become more pronounced, and severely itchy red or brown nodules (nodular pruritic rash, nodular prurigo) may appear, but are not limited to these.

[0055] As used herein, "urticaria" is a disease manifesting as an itchy rash and / or angioedema, and "chronic urticaria" is defined as urticaria present for six weeks or more, either continuously or intermittently.

[0056] As used herein, the term "asthma" refers to a disease in which the bronchi in the lungs become highly sensitive, causing occasional shortness of breath, a raspy sound in the breath, and severe coughing, and is an allergic disease caused by an allergic inflammatory reaction in the bronchi, and includes bronchial asthma, allergic asthma, and non-allergic asthma.

[0057] Allergic rhinitis and bronchial asthma are representative respiratory allergic diseases, with allergic rhinitis being an upper airway inflammatory disease and asthma being a lower airway inflammatory disease. Both diseases are IgE-mediated inflammatory diseases that cause airway hyperresponsiveness and can exhibit enhanced reactivity to irritants. At the same time, they share common inflammatory cells, including eosinophils, mast cells, and Th2 cells, and may produce common inflammatory factors such as histamine, leukotrienes, and Th2 cytokines. Therefore, the two diseases are closely related to each other.

[0058] Allergic rhinitis is one of the most common chronic inflammatory diseases in both adults and children. It is a condition characterized by a hypersensitive reaction of the nasal mucosa to various causative substances. Its main symptoms include a runny nose, nasal congestion, itchy nose, and cough, and may be accompanied by eye symptoms such as itchy and bloodshot eyes. Typical causes of allergic rhinitis include pollen, dust mites, and pet dander. Air pollution and sudden temperature changes can exacerbate symptoms. Well-known exacerbating factors include exercise, allergens or irritants, weather changes, and respiratory viral infections, including the common cold. Based on the results of several epidemiological and pathophysiological studies, the close association between the two diseases has been confirmed. Since then, the concept of "one airway disease" or "united disease" has been proposed, which considers the two allergic inflammatory diseases of the upper and lower respiratory tracts as a single airway disease rather than as separate entities. Therefore, it is important to determine whether patients with rhinitis also have asthma. Even in asthma patients, a comprehensive approach is recommended, including simultaneous evaluation of the presence or absence of rhinitis as a comorbidity.

[0059] The term "dust mite" as used herein refers to dust mites, whose corpses or feces can irritate the skin and mucous membranes, secreting histamine, a substance that causes inflammation in the body, and aggravating atopic dermatitis.

[0060] In addition to the effects of regulating, preventing, improving and treating allergic diseases as described above, the composition of the present invention induces the normalization of intestinal microorganisms altered by allergic diseases, thereby exhibiting excellent effects in regulating, preventing, improving and treating allergic diseases and their associated complications.

[0061] As used herein, the term "treat" refers to any act of ameliorating or beneficially altering the symptoms of an allergic disease by administering a pharmaceutical composition of the present invention.

[0062] As used herein, the term "prevent" refers to any action of suppressing the symptoms or slowing the progression of an allergic disease by administering the pharmaceutical composition of the present invention.

[0063] As used herein, the term "containing as an active ingredient" means that the strain, the lysate of the strain, the culture medium, or an extract of the culture medium is added to an extent that the effect can be exhibited, and also includes the addition of various components as accessory components for drug delivery, stabilization, etc., and formulation into various forms.

[0064] In one embodiment, the composition may contain 0.001% to 80% by weight of the Lactiplantibacillus plantarum strain based on the total weight of the composition. Furthermore, the dosage of the Lactiplantibacillus plantarum strain may be 0.01 mg to 10,000 mg, 0.1 mg to 1,000 mg, 1 mg to 100 mg, 0.01 mg to 1,000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. The strain is included in the composition at a therapeutically effective amount or a nutritionally effective concentration, for example, when the strain is 10 3 ~10 16 CFU / g, 10 3 ~10 15 CFU / g, 10 3 ~10 14 CFU / g, 10 3 ~10 13 CFU / g, 10 3 ~10 12 CFU / g, 10 4 ~10 16 CFU / g, 10 4 ~10 15 CFU / g, 10 4 ~10 14 CFU / g, 10 4 ~10 13 CFU / g, 10 4 ~10 12 CFU / g, 10 5 ~10 16 CFU / g, 10 5 ~10 15 CFU / g, 105 ~10 14 CFU / g, 10 5 ~10 13 CFU / g, 10 5 ~10 12 CFU / g, 10 6 ~10 13 CFU / g, 10 6 ~10 12 CFU / g, 10 7 ~10 13 CFU / g, 10 7 ~10 12 CFU / g, 10 8 ~10 13 CFU / g or 10 8 ~10 12 The composition may contain 1×10 CFU / g or a culture of the same number of live or dead bacteria. 3 ~1×10 16CFU / g of live or killed bacteria can be administered in a single dose or in divided doses. However, the dosage can be varied depending on factors such as the formulation method, administration method, the patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and those skilled in the art can appropriately control the dosage taking these factors into account. The number of administrations can be one or more times within the range of clinically acceptable side effects, and the administration can be performed at one or more sites. For non-human animals, the dosage can be the same as that for humans per kg (body weight), or an amount converted to the dosage based on, for example, the volume ratio (e.g., average value) of organs (e.g., heart) between the target animal and humans can be administered. Possible routes of administration include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intra-arterial, intraperitoneal, intradural, or intravenous), rectal, topical (including transdermal), inhalation, and injection, or insertion of an implantable device or substance. In one embodiment, target animals for treatment include humans and other mammals of interest, and specific examples include humans, monkeys, mice, rats, rabbits, sheep, cattle, dogs, horses, pigs, etc. According to one embodiment, the composition contains a killed dried strain of bacteria, and can be administered at a dose of 1 g to 10 g, 0.5 g to 1.5 g, 2.5 g to 3.5 g, or 4.5 g to 5.5 g per dose, and can be administered once to three times per day.

[0065] Another aspect provides a food composition comprising a lactiplantibacillus plantarum strain, a lysate derived from said strain, a culture broth, or a mixture thereof as an active ingredient.

[0066] Another aspect provides a health functional food for preventing or ameliorating allergic diseases, which contains a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0067] In one embodiment, the food composition or health functional food may have anti-allergic activity.

[0068] The "strain", "antiallergic activity", and "allergic disease" are as defined above.

[0069] In one embodiment, the improvement of the allergic disease can include a reduction in immune hypersensitivity reactions or an improvement in skin conditions caused by immune hypersensitivity reactions.

[0070] In one embodiment, the amelioration of the allergic disease can include suppressing Th2-mediated cytokine production, suppressing mast cell degranulation activity, or suppressing serum IgE or MCPT-1 levels.

[0071] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0072] In one embodiment, the allergic disease may be selected from the group consisting of edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic tympanitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, dry eye, pruritus, anaphylactic shock, hay fever, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0073] In one embodiment, the food composition may be for preventing or ameliorating an allergic disease.

[0074] In the food composition, the food includes meat, sausage, bread, chocolate, candy, snacks, confectionery, cereal, bakery products, dairy products, pizza, ramen, other noodles, gum, dairy products including ice cream, various types of milk powder, infant formula, milk, milk powder, yogurt, cheese, fermented milk, soup, beverages, tea, energy drinks, alcoholic beverages, multivitamin supplements, food additives, functional health foods and health foods, and includes all foods in the usual sense.

[0075] The term "functional food" is the same as "food for special health use" (FOSHU), and refers to a food with high medical and therapeutic effects that is processed to efficiently exert bioregulatory functions in addition to providing nutrients. Here, "functional" refers to obtaining beneficial health effects, such as regulating nutrients for the structure and function of the human body or achieving physiological effects. The food of the present application can be manufactured using methods commonly used in the art, and can be produced by adding raw materials and ingredients commonly used in the art. The food can also be manufactured in any dosage form that is recognized as a food. The food composition of the present application can be manufactured in various dosage forms, and unlike general pharmaceuticals, it has the advantage of being made from food ingredients, eliminating side effects that can occur with long-term drug administration. It is also highly portable, and HD-02 of the present application can be taken as an adjuvant to enhance the effects of preventing or improving allergies.

[0076] The term "health food" refers to food that has more active health maintenance and promotion effects than general foods, and "health supplement food" refers to food for the purpose of supplementing health. In some cases, the terms "health functional food," "health food," and "health supplement" can be used interchangeably.

[0077] Specifically, the functional health food is a food product produced by adding the composition according to one example to food ingredients such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, suspending, etc., and when ingested, it brings about specific health benefits. However, unlike general medicines, it has the advantage of being made from food ingredients and not causing side effects that can occur when taking medicines for a long period of time.

[0078] The food composition according to the example can be taken on a daily basis, and is therefore expected to be highly effective in preventing or improving allergic diseases, making it very useful.

[0079] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0080] The food composition may also contain additional ingredients commonly used in food compositions to improve aroma, taste, and visual appearance. Examples include vitamins A, C, D, E, B1, B2, B6, and B12, niacin, biotin, folate, and panthotenic acid. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr) may also be included. Amino acids such as lysine, tryptophan, cysteine, and valine may also be included.

[0081] The food compositions may also contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high-strength bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxycyanisol (BHA), butylhydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium D-bitartrate, etc.), fortifiers, emulsifiers, thickeners (thickeners), coating agents, gum bases, foam inhibitors, solvents, and improvers. The additives may be selected and used in appropriate amounts depending on the type of food.

[0082] The composition according to one embodiment may be added directly or in combination with other foods or food ingredients, and may be used as appropriate according to conventional methods. The amount of active ingredient to be added may be determined appropriately depending on the intended use (prevention, health, or therapeutic treatment). Generally, during the production of a food or beverage, the food composition of the present application may be added in an amount of 50 parts by weight or less, specifically 20 parts by weight or less, to the food or beverage. However, when ingested over a long period of time for health and hygiene purposes, a content below the above range may be included, and since there is no safety issue, the active ingredient may also be used in an amount greater than the above range.

[0083] An example of the food composition may be a health drink composition, which may contain various flavorings or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, or sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that may be used include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates per 100 mL of the health drink composition of the present application may generally be about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g.

[0084] In addition to the above, the health drink composition may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonation agents. Fruit pulp may also be included for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination. The proportion of such additives is not critical, but is typically selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health drink composition of the present application.

[0085] The food composition according to an example may contain HD-02 in various weight percentages as long as it can exhibit the effect of preventing or improving allergic diseases. For example, the food composition according to an example may contain HD-02 in the following amounts relative to the total weight of the food composition: 0.0001 to 100 wt%, 0.0001 to 80 wt%, 0.0001 to 50 wt%, 0.0001 to 40 wt%, 0.0001 to 20 wt%, 0.0001 to 10 wt%, 0.0001 to 5 wt%, 0.0001 to 2 wt%, 0.0001 The fermented milk may contain HD-02 at a concentration of 0.0001% to 0.01% by weight, 0.0001% to 0.5% by weight, 0.0001% to 0.2% by weight, 0.0001% to 0.1% by weight, 0.0001% to 0.05% by weight, 0.0001% to 0.02% by weight, 0.0001% to 0.01% by weight, 0.0001% to 0.005% by weight, 0.0001% to 0.002% by weight, or 0.0001% to 0.001% by weight, based on the total weight of the food composition. Specifically, foods and beverages can contain HD-02 in an amount of 0.0001% to 0.2% by weight, 0.0001% to 0.1% by weight, 0.0001% to 0.05% by weight, 0.0001% to 0.02% by weight, or 0.0001% to 0.01% by weight, based on the total weight of the food composition. Specifically, powdered or solid foods can contain HD-02 in an amount of 0.001% to 50% by weight, 0.001% to 30% by weight, 0.001% to 25% by weight, 0.001% to 20% by weight, or 0.001% to 10% by weight, based on the total weight of the food composition. The amount of the active ingredient to be mixed can be appropriately determined depending on the intended use (prevention, health, or symptom relief). For example, Lactipranchibacillus plantarum HD-02 can be added in an amount of 10% to 50% by weight, 0.001% to 30% by weight, 0.001% to 25% by weight, 0.001% to 20% by weight, or 0.001% to 10% by weight, based on the total weight of the food composition. 3 CFU / g ~10 16 It may be included at a CFU / g concentration, but is not limited thereto.

[0086] The functional health food composition can be used alone or in combination with other foods or food ingredients, and can be used according to conventional methods. The amount of active ingredient can be determined appropriately depending on the intended use (prevention, health, or therapeutic treatment). Generally, the composition of the present specification can be added in an amount of 15 parts by weight or less to the raw materials during food or beverage production. The type of functional health food is not particularly limited. Among the types of functional health foods, beverage compositions can contain various flavorings or natural carbohydrates as additional ingredients, just like regular beverages. Examples of natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that can be used include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. The health food composition may also contain nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, or combinations thereof. The health functional food composition may also contain natural fruit juice, fruit juice drink, fruit pulp for the production of vegetable drink, or combinations thereof.

[0087] Another aspect provides a pharmaceutical composition for treating or preventing an allergic disease, comprising a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof, as an active ingredient.

[0088] The "strain", "antiallergic activity", and "allergic disease" are as defined above.

[0089] In one embodiment, the pharmaceutical composition may have anti-allergic activity.

[0090] In one embodiment, the anti-allergic activity can include inhibiting degranulation in activated mast cells, specifically inhibiting the secretion of β-hexosaminidase, histamine, prostaglandins, and serine proteases in activated mast cells.

[0091] In one embodiment, the anti-allergic activity may include reducing serum IgE levels or reducing the binding of IgE to surface receptors on mast cells, and may further include suppressing serum MCPT-1 levels.

[0092] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0093] In one embodiment, the allergic disease may be selected from the group consisting of edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic tympanitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, dry eye, pruritus, anaphylactic shock, hay fever, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0094] The pharmaceutical composition may further comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and the lubricant may be magnesium stearate, talc, or a combination thereof. The carrier may be an excipient, disintegrant, binder, lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, anhydrous calcium phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

[0095] The pharmaceutical composition can be formulated into an oral or parenteral dosage form. The oral dosage form can be granules, powders, liquids, tablets, capsules, dry syrups, or a combination thereof. The parenteral dosage form can be an injection.

[0096] Another aspect provides a cosmetic composition for preventing or ameliorating allergic diseases, comprising a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0097] The "strain", "antiallergic activity", and "allergic disease" are as defined above.

[0098] In one embodiment, the allergic condition may be a skin allergic condition.

[0099] In one embodiment, the cosmetic composition may have anti-allergic activity.

[0100] The cosmetic composition may have the form of, for example, a softening lotion, a nourishing lotion, a massage cream, a nourishing cream, an essence, a mask sheet, a gel, an ampoule, or a skin adhesive type cosmetic formulation.

[0101] The ingredients contained in the cosmetic composition may include ingredients commonly used in cosmetic compositions in addition to the composition as an active ingredient, and may include conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0102] Another aspect provides a feed composition for preventing or ameliorating allergic diseases, which comprises a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0103] In one embodiment, the feed composition may have anti-allergic activity.

[0104] The "strain", "antiallergic activity", and "allergic disease" are as defined above.

[0105] The term "feed" may refer to any natural or artificial diet, meal, or component of a meal that is suitable for or intended to be eaten, ingested, and digested by an animal. The type of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include plant-based feeds such as grains, nuts, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, gourds, or grain by-products; protein-based feeds, lipid-free feeds, oils and fats, minerals, oils and fats, single-cell proteins, zooplankton, or animal feeds such as food and drink.

[0106] The feed composition can be prepared by adding the mixed strain composition in an appropriate effective concentration range according to various feed manufacturing methods known in the art, and can be used as a feed additive composition for the purpose of preventing or ameliorating allergic diseases.

[0107] When the composition is manufactured as a feed additive, it can be manufactured in the form of a highly concentrated liquid (20 to 90%), or in the form of a powder or granules. The feed additive can further contain one or more of the following: organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, etc.; phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphates (polymerized phosphates); and natural antioxidants such as polyphenols, catechin, α-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid. When manufactured as a feed, the composition can be formulated in a conventional feed form and can contain conventional feed ingredients.

[0108] The feed additives can be administered to animals alone or in combination with other feed additives in an edible carrier. Furthermore, the feed additives can be easily administered to animals as a top dressing, by directly mixing them into animal feed, or in a separate oral dosage form. When the feed additives are administered separately from animal feed, they can be combined with a pharmaceutically acceptable edible carrier to produce immediate-release or sustained-release dosage forms, as is well known in the art. Such edible carriers can be solid or liquid, such as corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When a solid carrier is used, the feed additive can be in the form of a tablet, capsule, powder, troche, or lozenge, or a finely dispersed top dressing. When a liquid carrier is used, the feed additive can be in the form of a soft gelatin capsule, or a syrup, suspension, emulsion, or solution. The feed and feed additives may also contain auxiliary agents such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters, etc. The feed additives can be used by adding them to animal feed by immersion, spraying, or mixing.

[0109] The feed or feed additive can be applied to diets for a variety of animals, including mammals, poultry, and fish, including, but not limited to, pigs, cows, horses, sheep, rabbits, goats, rodents, and laboratory rodents such as mice, hamsters, and guinea pigs, as well as pets (e.g., dogs and cats), poultry including chickens, turkeys, ducks, geese, pheasants, and quail, and fish including carp, crucian carp, and trout.

[0110] Another aspect provides a method of preventing, ameliorating, or treating a condition in an individual, comprising treating or administering to an individual in need thereof an effective amount of the composition.

[0111] In one embodiment, the individual's condition may be a disease or condition mediated by IgE (Immunoglobulin E), or a condition associated with an allergic disease.

[0112] Administration can be by methods well known in the art. Administration can be by any means, such as by intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous administration, directly to the individual. Administration can be systemic or local.

[0113] The individual may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat, and may be in need of amelioration of a condition associated with inflammation or a condition associated with a bacterial infection.

[0114] The administration may be such that the composition according to one embodiment is administered at a dose of 0.00001 mg to 1,000 mg per individual per day, for example, 0.00001 mg to 500 mg, 0.00001 mg to 100 mg, 0.00001 mg to 50 mg, 0.00001 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg. However, dosages can vary depending on factors such as formulation method, administration method, patient age, weight, sex, pathological condition, diet, administration time, administration route, excretion rate, and reaction sensitivity. Those skilled in the art can appropriately adjust dosages taking these factors into account. Administration can be once daily or twice or more times within the range of clinically acceptable side effects. Administration can be performed at one or more sites, daily or every 2 to 5 days, with a total administration period of 1 to 30 days. If necessary, the same treatment can be repeated at an appropriate time. For non-human animals, the same dosage per kg as for humans can be administered, or an amount converted to the dosage based on, for example, the volume ratio (e.g., average) of organs (e.g., heart) between the target animal and humans can be administered. [Effects of the Invention]

[0115] According to one embodiment, the strain and its derived disruption solution, culture solution, extract of the culture solution, or mixture thereof can be effectively used for the prevention, amelioration, or treatment of allergic diseases. [Brief explanation of the drawings]

[0116] [Figure 1] FIG. 1 is a graph showing the degranulation inhibitory effect of L. plantarum HD-02 culture supernatant on RBL-2H3 mast cells. [Figure 2]FIG. 2 is a graph showing the allergic reaction-suppressing effect of the L. plantarum HD-02 strain in a mouse model of anti-DNP IgE-induced passive cutaneous anaphylaxis. [Figure 3] FIG. 3 is a graph showing the ear edema-inhibitory effect of L. plantarum HD-02 strain in a mouse model of MC903-induced atopic dermatitis. [Figure 4] FIG. 4 is a graph showing the serum IgE-reducing effect of L. plantarum HD-02 strain in a mouse model of MC903-induced atopic dermatitis. [Figure 5] FIG. 5 is a graph showing the serum MCPT-1 reducing effect of L. plantarum HD-02 strain in a mouse model of MC903-induced atopic dermatitis. [Figure 6] FIG. 6 is a graph showing the effect of L. plantarum HD-02 strain in alleviating symptoms of atopic dermatitis in a mouse model of house dust mite-induced atopic dermatitis. [Figure 7] FIG. 7 is a graph showing the serum IgE-reducing effect of the L. plantarum HD-02 strain in a house dust mite-induced atopic dermatitis mouse model. [Figure 8] FIG. 8 is a graph showing the transepidermal water loss-reducing effect of L. plantarum HD-02 strain in a house dust mite-induced atopic dermatitis mouse model. [Figure 9] FIG. 9 is a graph showing the itching-relieving effect of the L. plantarum HD-02 strain in a house dust mite-induced atopic dermatitis mouse model. [Figure 10] FIG. 10 is a graph showing the effect of L. plantarum HD-02 strain in reducing the number of B cells, T cells, and total cells in the axillary lymph nodes in a house dust mite-induced atopic dermatitis mouse model. [Figure 11] FIG. 11 is a graph showing the effect of L. plantarum HD-02 strain in reducing Th1 and Th2 cells in the axillary lymph nodes in a house dust mite-induced atopic dermatitis mouse model. [Figure 12]FIG. 12 is a graph showing the effect of L. plantarum HD-02 strain in reducing skin thickness in a house dust mite-induced atopic dermatitis mouse model. [Figure 13] FIG. 13 is a graph showing the inhibitory effect of L. plantarum HD-02 strain on mast cell infiltration and degranulation in a house dust mite-induced atopic dermatitis mouse model. DETAILED DESCRIPTION OF THE INVENTION

[0117] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided to facilitate understanding of the present invention, and the contents of the present invention are not limited to the following embodiments. Various modifications can be made to the embodiments, and the embodiments are not limited to the embodiments disclosed below, and can be implemented in various forms.

[0118] Example 1. Isolation and identification of Lactiplantibacillus plantarum strains

[0119] 1.1. Strain isolation Various types of kimchi grown at home were used to isolate Lactiplantibacillus plantarum strains. 25 g of kimchi sample was diluted in 225 ml of 0.85% NaCl solution, placed in a wire filter sample bag, and homogenized using a stomacher for 5 minutes. The filtrate was diluted 10-fold and smeared on MRS plates (pH 5.5, Difco, USA) and anaerobically incubated at 37°C for 48 hours. After incubation, single colonies were identified by morphology and color, and pure isolates were obtained through three subcultures. Strains that tested positive and negative for Gram staining and catalase testing were selected and arbitrarily named HD-02. Lactiplantibacillus plantarum HD-02 (hereinafter referred to as "HD-02") was finally selected.

[0120] 1.2. Molecular identification of selected strains To analyze the 16S rRNA base sequence of the HD-02 strain, the 16S rRNA gene was amplified using bacterial universal primers and then analyzed for base sequence (Macrogen Inc., Korea). PCR was performed using the primer sequences listed in Table 1, and the analytical result was the same as SEQ ID NO: 1.

[0121] [Table 1] The 16S rRNA sequences of each strain were analyzed using the sequence comparison program (BLAST) of the National Center for Biotechnology Information (NCBI). As a result, it was confirmed that the 16S rRNA gene of the HD-02 strain was more than 99% identical to that of several strains belonging to the Lactiplantibacillus plantarum species, including the reference strain of Lactiplantibacillus plantarum (JCM 1149) (Table 2).

[0122] [Table 2] Since the 16S rRNA sequence of Lactiplantibacillus plantarum is more than 99% identical to that of Lactiplantibacillus pentosus and other mycelia, we analyzed the recA gene, a taxonomic biomarker other than the 16S rRNA gene, for more accurate identification. Gene amplification was performed using the recA-specific primers listed in Table 3, and the base sequence was analyzed (Bionics Co., Ltd., Korea). The analytical result is the same as SEQ ID NO: 2.

[0123] [Table 3] The recA gene sequences of each strain were analyzed using the sequence comparison program (BLAST) of the National Center for Biotechnology Information (NCBI). The results showed that the HD-02 strain had 99% identity with the recA gene of Lactiplantibacillus plantarum (Table 4).

[0124] [Table 4]

[0125] 1.3. Morphological and physiological characterization of selected strains The morphological and physiological characteristics of the HD-02 strain were determined using Bergy's manual of systematic bacteriology and the API50CHL kit (Biomerieux SA, France). The morphological and physiological characteristics of the HD-02 strain are summarized in Table 5, and the sugar fermentation characteristics of the strain are summarized in Table 6.

[0126] [Table 5]

[0127] [Table 6-1] [Table 6-2] Fermentation characteristics evaluation on a positive (+) / negative (-) basis showed that the HD-02 strain showed 66.8% similarity to Lactiplantibacillus plantarum.

[0128] The strain isolated based on the results of the 16S rRNA and recA gene sequence analyses and morphological and physiological characteristics was named Lactiplantibacillus plantarum HD-02, and the HD-02 strain was deposited at the Korea Advanced Industrial Genetic Resource Center (KACC) on August 26, 2022 (accession number: KACC81228BP).

[0129] Experimental Example 1. Degranulation inhibitory effect of L. plantarum HD-02 culture supernatant on RBL-2H3 mast cells Culture supernatants of various L. plantarum strains, including L. plantarum HD-02, were treated with activated mast cells that cause allergic reactions, and their inhibitory effects on degranulation were screened by measuring β-hexosaminidase secretion.

[0130] RBL-2H3 cells were suspended in 15% FBS and MEM medium in a 24-well plate, and 1.5 x 10 cells were dispensed per well. They were then sensitized with 20 ng / mL DNP-IgE for approximately 12 hours. After washing once with PIPES buffer (25 mM PIPES, 119 mM NaCl, 5 mM KCl, 0.4 mM MgCl2, 1 mM CaCl2, 5.6 mM glucose, 0.1% BSA), culture supernatants of various L. plantarum strains were diluted 1:15 in PIPES buffer and pretreated in each well for 2 hours. Culture supernatants were prepared by culturing L. plantarum strains in MRS medium, centrifuging the cells, and then filtering the supernatant through a 0.22 μm filter. Cells were then stimulated with 25 ng / mL DNP-HSA (antigen) for 15 minutes, followed by cooling on ice for 5 minutes to terminate the reaction. The supernatant was transferred to an e-tube and washed once with PIPES buffer. The remaining cells were then disrupted with 0.1% Triton X-100. The supernatant and lysed cell samples were mixed in 30 μL portions with substrate buffer (1 mM 4-nitrophenyl N-acetyl-β-D-glucosaminide, 0.1 M sodium citrate) in a 96-well plate and incubated at 37°C for 1 hour. The reaction was terminated by adding 0.1 M carboxylic acid buffer, and the absorbance was measured at 405 nm using a microplate reader. The results are shown in Figure 1.

[0131] As shown in Figure 1, after screening the culture supernatants of 14 different L. plantarum strains, we confirmed that the culture supernatant of the L. plantarum HD-02 strain had the greatest inhibitory effect on mast cell degranulation.

[0132] Experimental Example 2. Inhibitory effect of L. plantarum HD-02 on IgE-mediated allergic reactions in an animal model of passive cutaneous anaphylaxis The inhibitory effect of L. plantarum HD-02 strain on the allergic reaction was confirmed in an animal model of passive cutaneous anaphylaxis (PCA) induced by anti-DNP IgE.

[0133] Four-week-old BALB / c mice were obtained, and after a one-week purification period, freeze-dried L. plantarum HD-02 strain was suspended in D-PBS and administered at a dose of 1 × 10 per mouse from 2 days before IgE sensitization until just before the end of the test. 10 The mice were orally administered daily at a concentration of 1000 CFU. Dexamethasone (Dexa) was used as a positive control at a concentration of 10 mg / kg. To induce passive skin sensitization, 20 ng of anti-DNP IgE was injected topically into one ear of each mouse. Approximately 24 hours later, DNP-HSA was diluted to 0.5 mg / mL in 5 mg / mL Evans Blue dye solution and 200 μL was administered via the tail vein. After 30 minutes, the mice were euthanized, and the ears were removed and placed in 700 μL of formamide. The dye exuded into the ear tissue was extracted after 12 hours at 63°C. The absorbance of the supernatant was measured at 620 nm. Quantitation of exuded Evans Blue was measured using a standard curve, as shown in Figure 2. As shown in Figure 2, the anti-allergic effect was confirmed by suppressing the allergic reaction by approximately 20.4% in the experimental group (Ag+HD-02) administered with the L. plantarum HD-02 strain compared to the negative control group (Ag) in a passive skin sensitization allergic reaction model using anti-DNP IgE.

[0134] Experimental Example 3. Inhibitory effect of L. plantarum HD-02 on allergic reactions in an animal model of MC903-induced atopic dermatitis

[0135] 3.1. Inhibitory effect of L. plantarum HD-02 on ear edema The effect of L. plantarum HD-02 strain in suppressing ear edema was confirmed in an animal model of MC903-induced atopic dermatitis. Six-week-old BALB / c mice were obtained. After a one-week purification period, freeze-dried L. plantarum HD-02 strain was suspended in D-PBS and inoculated at 2 × 10 per mouse from 7 days before the induction of atopic dermatitis until just before the end of the study. 9After the mice were restrained to induce atopic dermatitis, the disease-inducing group received 2 nmol MC903 (calcipotriol, C) dissolved in ethanol in both ears. 27 H 40 20 μL of each of the ethanol-containing ethanol (O3) was applied daily for 12 days, while the control group received only ethanol. To assess the degree of ear edema due to atopic dermatitis induction, the ear thickness of the mice was measured at the same time each day, as shown in Figure 3. The change in ear edema was calculated by subtracting the ear thickness measured before induction from the ear thickness measured daily after induction. As shown in Figure 3, in the MC903-induced atopic dermatitis model, the experimental group (MC903+HD-02) administered with L. plantarum HD-02 strain showed an approximately 48.4% ear edema suppression effect compared to the control group (MC903).

[0136] 3.2. Reduction of serum total IgE in the L. plantarum HD-02 strain-administered group Overexpression of IgE is the most representative marker of atopic dermatitis, and the total serum IgE concentration was measured using a mouse IgE ELISA kit (Biolended #432404) to confirm the serum IgE concentration in the MC903-induced atopic dermatitis animal model described in Experimental Example 3.1 above.

[0137] The microwell plate was incubated overnight at 4°C with a capture antibody diluted in coating buffer, washed four times with wash buffer, and blocked with assay diluent for 1 hour at room temperature. After washing four times, a standard sample and a serum sample diluted 1:200 were added and incubated for 2 hours at room temperature. After washing four times, the plate was incubated with a detection antibody for 1 hour at room temperature. After washing four times, avidin-HRP was added and incubated for 30 minutes at room temperature. After washing five times, TMB substrate solution was added and incubated for 20 minutes at room temperature in the dark. The color reaction was stopped by adding stop solution to each well, and the absorbance was measured at 450 nm using an ELISA reader. The results are shown in Figure 4.

[0138] As shown in Figure 4, the control group (MC903) showed a rapid increase in serum total IgE concentration compared to the normal group, but the experimental group (MC903+HD-02) administered with the L. plantarum HD-02 strain showed a reduction in serum total IgE concentration of approximately 57.2%.

[0139] 3.3. Serum MCPT-1 reduction effect in the L. plantarum HD-02 strain administration group To confirm the serum mast cell protease-1 (MCPT-1) concentration in the MC903-induced atopic dermatitis animal model in Experimental Example 3.1, a mouse MCPT-1 (mMCP-1) ELISA kit (Invitrogen #88-7503) was used to measure it.

[0140] The microwell plate was incubated overnight at 4°C with a capture antibody diluted in coating buffer, washed three times with wash buffer, and blocked with ELISA / ELISPOT diluent for 1 hour at room temperature. After washing three times, a standard sample and a serum sample diluted 1:100 were added and incubated for 2 hours at room temperature. After washing three times, the detection antibody was added and incubated for 1 hour at room temperature. After washing three times, avidin-HRP was added and incubated for 30 minutes at room temperature. After washing five times, TMB substrate solution was added and incubated for 15 minutes at room temperature in the dark. The color reaction was stopped by adding stop solution to each well, and the absorbance was measured at 450 nm using an ELISA reader. The results are shown in Figure 5.

[0141] As shown in Figure 5, the control group (MC903) showed a rapid increase in serum MCPT-1 concentration compared to the normal group, but the experimental group (MC903+HD-02) administered with the L. plantarum HD-02 strain showed a reduction in serum MCPT-1 concentration of approximately 56.8%.

[0142] Experimental Example 4. Inhibitory effect of L. plantarum HD-02 on allergic reactions in an animal model of house dust mite-induced atopic dermatitis

[0143] 4.1. Effect of L. plantarum HD-02 on atopic dermatitis symptoms The suppressive effect of L. plantarum HD-02 strain on atopic dermatitis symptoms was confirmed in an animal model of house dust mite-induced atopic dermatitis.

[0144] Six-week-old NC / Nga mice were obtained and, after a one-week depilation period, the mice's backs and the area above the ears were thoroughly depilated. To remove cutaneous fat and disrupt the cuticle barrier, 150 μL of 4% SDS aqueous solution was sprayed onto the depilated area, allowed to dry completely, and then 100 mg of Biosta AD (a natural mite-derived ingredient, Dermatophagoides farinae) ointment was applied evenly. The inducer was applied twice a week for three weeks to induce atopic dermatitis, and then once a week for eight weeks to maintain the condition. Administration of the L. plantarum HD-02 strain was initiated upon completion of the three-week atopic dermatitis induction period and continued until just before the end of the study, at a dose of 2 × 10 per mouse. 9 The mice were orally administered 100 μL of dexamethasone at a concentration of 0.1% twice a week to each individual, which served as a positive control.

[0145] The severity of atopic dermatitis symptoms was evaluated weekly as a Dermatitis Score, which was a composite score of the following four items: erythema / hemorrhage, scarring / dryness, edema, and excoria / erosion. Each item was scored as 0 (no symptoms), 1 (weak), 2 (normal), or 3 (severe), and the scores were then summed. The results are shown in Figure 6.

[0146] As shown in Figure 6, the severity of atopic dermatitis in the control group (dust mite induction) was measured at an average of 8.17 points, while the score for the positive control group (Dexa) was measured at 2.50, demonstrating an approximately 69.4% alleviation of atopic dermatitis symptoms compared to the control group. Furthermore, the severity of atopic dermatitis in the experimental group (dust mite induction + HD-02) was measured at an average of 3.00 points, demonstrating an approximately 63.3% alleviation of atopic dermatitis symptoms, which was confirmed to be at a similar level to the positive control group (Dexa).

[0147] 4.2. Effect of L. plantarum HD-02 strain administration on serum total IgE levels In order to confirm the serum total IgE concentration in the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1, measurements were performed using a mouse IgE ELISA kit in the same manner as in Experimental Example 3.2, and the results are shown in Figure 7.

[0148] As shown in Figure 7, the control group (dust mite induction) showed a rapid increase in serum total IgE concentration compared to the normal group (dust mite induction X). Compared to the control group (dust mite induction), the positive control group (Dexa) and the experimental group (dust mite induction + HD-02) showed a suppression effect on serum total IgE of approximately 63.7% and approximately 50.6%, respectively.

[0149] 4.3. Transepidermal water loss reduction effect of L. plantarum HD-02 strain administration group In order to confirm transepidermal water loss due to damage to the skin barrier function in the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) in Experimental Example 4.1, the water evaporation rate on the skin surface was measured using a Tewameter™ Nano device, and the results are shown in Figure 8.

[0150] As shown in Figure 8, the control group (dust mite-induced) showed a significant increase in transepidermal water loss compared to the normal group (dust mite-induced X). Compared to the control group, the positive control group (Dexa) showed a decrease of approximately 51.3%, and the experimental group (dust mite-induced + HD-02) showed a decrease of approximately 36.9% in transepidermal water loss. This confirmed the skin barrier-improving effect of the L. plantarum HD-02 strain.

[0151] 4.4. Itching relief effect in the L. plantarum HD-02 strain-administered group In the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) in Experimental Example 4.1, itching was confirmed, which can demonstrate the degree of itching at the atopic skin lesion site, an important evaluation index of the disease model. Mice in all groups were photographed for 10 minutes, and the frequency of scratching behavior of the photographed mice was measured. The results are shown in Figure 9.

[0152] As shown in Figure 9, the control group (dust mite induction) showed an approximately 18.7-fold increase in the number of scratching episodes compared to the normal group (dust mite induction X), and the positive control group (Dexa) and the experimental group (dust mite induction + HD-02) showed an itching relief effect of approximately 51.5% and 55.2%, respectively, compared to the control group.

[0153] 4.5. Confirmation of changes in cell composition in the axillary lymph nodes of the L. plantarum HD-02 strain-administered group From the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1, changes in immune cell composition in the axillary lymph node, which is an important immune tissue in the disease model, were confirmed.

[0154] Axillary lymph nodes were removed from each mouse to obtain a single cell suspension, and the cell number was measured to obtain 1.0 × 10 cells per well. 6The cells were aliquoted into a 96-well U-low microwell plate at a cell count of 1000. After 4 hours of stimulation with eBioscience® Cell Stimulation Cocktail (plus protein transport inhibitors), the cells were stained with anti-CD3-Pacific blue, anti-CD4-PE-Cy7, anti-IL-10-FITC, anti-IL-4-PE, and anti-IFN-γ-APC antibodies and subjected to FACS analysis. The results are shown in Figures 10 and 11.

[0155] As shown in Figure 10, the total cell count in the axillary lymph nodes of the atopic dermatitis-induced control group (dust mite induction) was approximately 7.5-fold higher than that of the normal group (dust mite induction X), and the axillary lymph node cell count was reduced by approximately 81.7% in the positive control group (Dexa) and approximately 53.7% in the experimental group (dust mite induction + HD-02) compared to the control group. Furthermore, as shown in Figure 11, administration of the L. plantarum HD-02 strain also significantly reduced B cells, T cells, Th1 and Th2 cells in the axillary lymph nodes compared to the control group.

[0156] 4.6. Confirmation of skin thickness and histological changes in the L. plantarum HD-02 strain-administered group The effectiveness of L. plantarum HD-02 strain on the increase in skin thickness and mast cell infiltration associated with the inflammatory response was confirmed using the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1.

[0157] The skin from the backs of mice was removed and fixed in 10% formalin. Paraffin blocks were then prepared and sliced ​​to a thickness of 4 μm to create slides. Hematocillin-eosin (H&E) staining was used to observe lymphocyte accumulation in the skin tissue and changes in epidermal thickness, and toluidine blue staining was used to measure the number of mast cells and confirm the degree of tissue infiltration. The results are shown in Figures 12 and 13.

[0158] As shown in Figure 12, the atopic dermatitis-induced control group (dust mite induction) showed an approximately 8.8-fold increase in epidermal layer thickness compared to the normal group (dust mite induction X), and the positive control group (Dexa) and the experimental group (dust mite induction + HD-02) showed a reduction in epidermal layer thickness of approximately 56.7% and 57.3%, respectively, compared to the control group. Furthermore, the experimental group showed a significant reduction in lymphocyte accumulation in inflamed tissue at the same level as the positive control group (Dexa).

[0159] Furthermore, as shown in Figure 13, the infiltration of mast cells into inflamed tissue was confirmed, and the number of mast cells in the control group in which atopic dermatitis was induced increased by approximately 6.2 times compared to the normal group, and the number of mast cells was significantly reduced by approximately 61.4% in the positive control group (Dexa) and approximately 47.0% in the experimental group compared to the control group.

[0160] These results indicate that the L. plantarum HD-02 strain can be useful for preventing, ameliorating, or treating atopic dermatitis or allergic diseases.

[0161] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and are not limiting. [Accession number]

[0162] Depository institution: Agricultural Genetic Resource Center (overseas) Accession number: KACC81228BP Date of acceptance: 20220826 TIFF2026502882000009.tif204170

Claims

1. An antiallergic composition comprising, as an active ingredient, Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC 81228BP, a disruption solution derived from said strain, a culture solution derived from said strain, or a mixture thereof.

2. 2. The antiallergic composition according to claim 1, which suppresses the production of Th2 (helper T2 cell)-mediated cytokines.

3. 2. The antiallergic composition according to claim 1, which reduces serum IgE levels or reduces the binding of IgE to surface receptors on mast cells.

4. 10. The antiallergic composition of claim 1, further comprising one or more additives selected from the group consisting of preservatives, dyes, emulsifiers, sweeteners, stabilizers, flavor enhancers, flavoring agents, and acidulants.

5. 2. The antiallergic composition according to claim 1, which is in a dosage form selected from the group consisting of a solution, an emulsion, a viscous mixture, a powder, a granule, a tablet, and a capsule.

6. A food composition comprising, as an active ingredient, Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a disruption liquid derived from said strain, a culture liquid derived from said strain, or a mixture thereof.

7. 7. The food composition of claim 6, selected from the group consisting of ice cream, infant formula, milk, yogurt, cheese, fermented milk, powdered milk, cereal, bakery products, dairy products, beverages, and food additives.

8. A pharmaceutical composition for treating or preventing an allergic disease, comprising the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a lysate derived from said strain, a culture broth derived from said strain, or a mixture thereof, as an active ingredient.

9. The pharmaceutical composition according to claim 8, which has an inhibitory activity against mast cell degranulation.

10. The pharmaceutical composition according to claim 8, which i) suppresses the expression of IgE, or ii) suppresses the level of MCPT-1 in serum.

11. The pharmaceutical composition according to claim 8, wherein the allergic disease comprises an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

12. 9. The pharmaceutical composition according to claim 8, wherein the allergic disease is selected from the group consisting of edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic otitis media, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, dry eye, pruritus, anaphylactic shock, hay fever, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

13. A health functional food for improving or preventing allergic diseases, comprising, as an active ingredient, Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a lysate derived from said strain, a culture broth derived from said strain, or a mixture thereof.

14. The improvement of the allergic disease is achieved by the following: - suppression of Th2-mediated cytokine production, - Inhibition of mast cell degranulation activity, - suppression of serum IgE expression, or - suppression of serum MCPT-1 levels, The health functional food according to claim 13, comprising:

15. The health functional food according to claim 13, wherein the improvement of the allergic disease includes a reduction in immune hypersensitivity reaction or an improvement in skin conditions caused by immune hypersensitivity reaction.

16. The health functional food according to claim 13, wherein the allergic disease includes an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

17. The allergic diseases include edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic otitis media, atopic dermatitis, contact dermatitis, urticaria, and chronic spontaneous urticaria. urticaria), dry eye, pruritus, anaphylactic shock, hay fever, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy and respiratory allergy.

18. A cosmetic composition for improving or preventing skin allergies, comprising the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a homogenate derived from said strain, a culture broth derived from said strain, or a mixture thereof.

19. A feed composition for improving or preventing allergic diseases, comprising the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a lysate derived from said strain, a culture broth derived from said strain, or a mixture thereof.

20. A method for preventing or treating an allergic disease, comprising the step of administering to an individual in need thereof an effective amount of the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a culture of said strain, a lysate of said strain, or a composition comprising a mixture thereof.

21. Use of a composition comprising the Lactiplantibacillus plantarum HD-02 strain deposited under accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof, for the manufacture of a pharmaceutical preparation for preventing or treating an allergic disease.

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