Lactiplantibacillus plantarum strain, culture medium derived therefrom and its anti-allergic use

Abstract

With regard to novel microorganisms, their disrupted products, culture broth, extracts of the culture broth, and their anti-allergic uses, the novel strains according to one embodiment, their derived disrupted products, culture broth, extracts of the culture broth, or mixtures thereof, are effective in preventing, ameliorating, or treating allergic diseases.

Description

[Technical Field]

[0001] The present invention relates to a novel microorganism, its disrupted product, culture medium, extract of the culture medium, and antiallergic use thereof. [Background technology]

[0002] The microbiome refers to the microorganisms that exist in a specific environment and their entire genetic information, and is a collection of genomes, which represent the entire genetic information of a single organism. Therefore, the human microbiome refers to the microorganisms that live inside and outside the human body and their entire genetic information.

[0003] The human body lives in a symbiotic relationship with many microorganisms, and the intestines in particular are the ideal environment for microorganisms to obtain nutrients and form systematic communities, with the greatest number of microorganisms present. Gut microorganisms supply nutrients that the host cannot produce using its own enzymes alone, and are closely related to the host's metabolism and immune system. They have also been reported to be associated with the development of various diseases, including irritable bowel syndrome, obesity, atopy, depression, rheumatoid arthritis, autism spectrum disorder, and dementia.

[0004] Allergies include not only IgE-mediated allergic hypersensitivity reactions but also non-IgE-mediated hypersensitivity reactions. IgE-mediated allergic diseases are caused by IgE allergy antibodies, while non-IgE-mediated allergic diseases are caused by reactions involving other components of the immune system. The mechanism of non-IgE-mediated allergic diseases is not well understood, but it is known to involve innate immunity and cellular immunity (T lymphocyte processes), with no substantial involvement of IgE antibodies. Mast cell activation induces degranulation, resulting in the extracellular secretion of chemical mediators such as histamine, cytokines, and chemokines, which induce increased vascular permeability, smooth muscle contraction, and increased mucus secretion, resulting in type I allergic reactions.

[0005] Mast cells produce various cytokines via their high-affinity IgE receptors (FcεRI), which promote the expression of inflammatory factors, leading to inflammatory allergic reactions. Among these, IL-4 is produced by T lymphocytes and mast cells, induces the production of IgE, which induces type I allergic reactions, and regulates the mediated immune response. Increased IL-4 production has been reported in actual patients (Ngoc et al., 2005). Furthermore, when an allergic reaction begins in the body, TNF-α is degranulated from mast cells and released, along with histamine and other inflammatory mediators, to induce the production of other cytokines and inflammatory mediators, further exacerbating the allergic and inflammatory response (Chung & Barnes, 1999). Histamine is present prior to an allergic reaction and is released externally upon exposure to an allergic stimulus (Schwartz & Huff, 1998).

[0006] Allergies are a malfunction of the immune system, causing hypersensitivity reactions such as hives, itching, runny nose, and coughing in certain individuals, even to substances that normally have little effect on humans. They can be caused by environmental factors such as environmental pollution, westernized diet and lifestyle, and stress, as well as genetics and various allergens. The prevalence of allergies is increasing worldwide, with 20-25% of the population suffering from allergic diseases such as rhinitis, asthma, atopic dermatitis, and food allergies. Recent evidence suggests that allergies are also associated with late-stage and chronic allergic reactions, indicating their involvement in allergic inflammation in general.

[0007] Various methods have been proposed to treat allergic diseases, including allergen avoidance, anti-allergy drug administration, regulation of IgE synthesis in the body, and development of anti-IgE antibodies. However, these methods have many drawbacks, such as failure to treat the underlying cause of allergies, insufficient efficacy of drugs, and the occurrence of serious side effects.

[0008] Therefore, there is an urgent need to discover substances with anti-allergic activity for the development of allergy treatments. To be considered to have anti-allergic activity, it is not enough to simply have therapeutic activity against IgE-mediated allergies; it must also have therapeutic activity against non-IgE-mediated allergies.

[0009] In recent years, as a result of the progress of metagenomics projects to elucidate the intestinal bacterial flora, correlations between the distribution of intestinal flora and various immune system diseases, including allergies, have been revealed, and it is becoming clear that the distribution and diversity of intestinal bacteria, as well as intestinal immunity and the immune system, are closely related. Utilizing the results of elucidating the intestinal bacterial flora, it is necessary to develop substances that have therapeutic activity not only for IgE-mediated allergies but also for non-IgE-mediated allergies. Summary of the Invention [Problem to be solved by the invention]

[0010] One aspect is to provide a composition for suppressing the production of Th2 (helper T2 cell)-mediated cytokines or alleviating allergic reactions, comprising a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus as an active ingredient.

[0011] One aspect provides a strain of Lactiplantibacillus plantarum GBCC_F0070 belonging to the genus Lactiplantibacillus, deposited under accession number KCTC15051BP.

[0012] Another aspect is to provide a lysate, culture medium, or culture medium extract from said strain.

[0013] 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.

[0014] Another aspect is to provide a pharmaceutical composition for treating or preventing allergic diseases, comprising a lactiplantibacillus plantarum strain, a lysate or culture medium derived from said strain, or a mixture thereof, as an active ingredient.

[0015] Another aspect is to provide a cosmetic composition for preventing or improving skin allergies, which contains a lactiplantibacillus plantarum strain, a lysate or culture solution derived from said strain, or a mixture thereof as an active ingredient.

[0016] Another aspect provides a method for preventing or treating an allergic disease, comprising the step of administering to an individual in need thereof an effective amount of a composition comprising the Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, a culture of said strain, a homogenate of said strain, or a mixture thereof.

[0017] Another aspect provides use of a composition comprising the Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, a culture of the strain, a disrupted product of the strain, or a mixture thereof, for the manufacture of a pharmaceutical preparation or a functional health food preparation for preventing or treating allergic diseases. [Means for solving the problem]

[0018] One aspect provides a composition for suppressing the production of Th2 (helper T2 cell)-mediated cytokines or alleviating allergic reactions, comprising a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus as an active ingredient.

[0019] One embodiment provides a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus.

[0020] 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 platarum GBCC_F0070 as a candidate anti-allergy strain. This strain was deposited at the Korea Institute of Bioscience and Biotechnology (KIBIT) Biological Resources Center on August 16, 2022, under accession number KCTC15051BP. This strain corresponds to a probiotic strain and is harmless to the human body, allowing it to be used without side effects.

[0021] In one embodiment, the Lactiplantibacillus plantarum strain may be the strain deposited under accession number KCTC15051BP.

[0022] In one embodiment, the Lactiplantibacillus plantarum strain may be a strain comprising the 16S rRNA sequence of SEQ ID NO:1.

[0023] 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.

[0024] In one embodiment, the Lactiplantibacillus plantarum strain may be a strain comprising the recA gene sequence of SEQ ID NO:2.

[0025] 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 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9%, or 100% homology to the gene sequence of SEQ ID NO: 2 herein.

[0026] In one embodiment, the strain may be a mutant strain of a naturally occurring strain.

[0027] In one embodiment, the bacterial strain may be live, killed, attenuated, or a cytoplasmic fraction obtained by disrupting the strain, preferably live.

[0028] In one embodiment, the strain may have anti-allergic activity. Specifically, without being limited to a particular theory, the mechanism of allergy induction can be divided into one stage and two stages.

[0029] In the first stage, external allergens penetrate the body through epithelial cells. The penetrated allergens are then ingested by antigen-presenting cells, which then present fragments to T cells. The antigen-presented T cells differentiate into Th2 (helper T2) cells and produce Th2-mediated cytokines such as interleukin-4 (IL-4). These cytokines activate B cells, inducing the production of immunoglobulin E (IgE) from B cells. The produced IgE antibodies then bind to FcεRI receptors present on the surface of mast cells. This state is called the atopic state, a state of readiness (standby) for allergy induction.

[0030] The second step involves the re-infiltration of allergens, which have been memorized by immune cells after the first step, and cross-linking with IgE bound to mast cells, inducing the degranulation of mast cells. This may result in the release of allergic inflammatory factors, such as histamine, leukotrienes, and tryptase, from the mast cells. These substances, including histamine, may be the direct cause of various allergic inflammatory diseases. As mentioned above, the current mechanisms of action of therapeutic agents and allergy-ameliorating substances may focus on preventing the release of histamine from mast cells, which is the final step in allergy, and the role of histamine.

[0031] 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.

[0032] In one embodiment, the strain may reduce serum IgE levels or reduce the binding of IgE to mast cell surface receptors, and may suppress serum MCPT-1 levels.

[0033] Therefore, the strain according to one embodiment can be useful in treating allergic diseases.

[0034] Another embodiment provides a lysate, culture broth, culture broth extract, or mixture thereof from a strain of Lactiplantibacillus plantarum.

[0035] The term "culture medium" as used herein can be used interchangeably with "culture supernatant," "culture supernatant," "conditioned culture medium," or "conditioned medium," and can refer to the entire medium containing the Lactiplantibacillus strain, its metabolites, extra nutrients, etc., obtained by culturing the strain for a certain period of time in a medium capable of supplying 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.

[0036] 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.

[0037] 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 Lactiplantibacillus 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 cellular debris, which may be smooth or granular in structure.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] In one embodiment, the composition may have anti-allergic activity.

[0045] In one embodiment, the anti-allergic activity may be suppression of Th2-mediated cytokine production, suppression of mast cell degranulation activity, suppression of serum IgE levels, suppression of serum MCPT-1 levels, or food allergy suppression activity. Specifically, the suppression of mast cell degranulation activity may be suppression of β-hexosaminidase secretion in activated mast cells.

[0046] In one embodiment, the Th2-mediated cytokine may be at least one selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

[0047] In one embodiment, the composition may reduce serum IgE levels or reduce the binding of IgE to mast cell surface receptors, and the strain may suppress serum MCPT-1 levels.

[0048] According to one embodiment, the anti-allergic activity may include the activity of inhibiting mast cell degranulation by 50% to 70% when the supernatant of Lactipranchi Bacillus plantarum GBCC_F0070 strain is treated in mast cells RBL-2H3.

[0049] According to one embodiment, the anti-allergic activity may include the activity of suppressing ear edema by 30% to 50% when the supernatant of the lactiprancid Bacillus plantarum GBCC_F0070 strain is administered to an animal model of MC903-induced atopic dermatitis.

[0050] According to one embodiment, the antiallergic activity may include the activity of reducing serum IgE levels by 50% to 70% or the activity of reducing serum MCPT-1 levels by 50% to 70% when the supernatant of Lactiplantibacillus plantarum GBCC_F0070 strain is administered to an MC903-induced atopic dermatitis animal model.

[0051] Another aspect provides a disease-modifying, preventative or therapeutic use of Lactiplantibacillus plantarum, a lysate, a culture broth or an extract of a culture broth of said strain.

[0052] In one embodiment, the use may include the prevention, amelioration, or treatment of a disease or condition mediated or not mediated by IgE (Immunoglobulin E). Specifically, the disease mediated or not mediated by IgE (Immunoglobulin E) may be an allergic disease.

[0053] As used herein, the term "allergic disease" refers to a disease caused by an excessive immune response in the body to an external antigen, specifically, edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, pruritus, anaphylactic shock, insect allergy, food allergy, drug allergy, and respiratory allergy, but is not limited to this.

[0054] The development of allergic diseases can be broadly divided into an early reaction and a late reaction. In the early reaction, after an antigen stimulates the body and produces an antibody (IgE), the antibody binds to a high-affinity receptor (FcεRI) on the surface of mast cells. When the antigen re-enters the body, it binds to the antibody (IgE) bound to FcεRI, rapidly transmitting a signal to the mast cells, causing the secretion of inflammatory chemical mediators (cytokines, histamine, leukotrienes) that cause allergies.

[0055] In the late-stage response, Th2-type 2 cytokines (IL-4, IL-5, IL-6, IL-10, IL-13) induce the production of chemokines (monocyte chemoattractant protein-1, eotaxin-1, RANTES) in tissue fibroblasts or epithelial cells, resulting in the migration of inflammatory cells, particularly eosinophils, to the site of the allergic reaction. As a result, many allergic symptoms manifest as clinical symptoms such as itching, edema, congestion, coughing, and phlegm, caused by inflammatory response mediators such as histamine secreted by mast cells and eosinophils, which dilate capillaries around epithelial cells in the airways and skin, influx of immune cells, muscle contraction, and increased mucus secretions.

[0056] In particular, among the Th2-type 2 cytokines, IL-4 is produced by T cells and mast cells and plays a role in regulating B cell class switching and promoting IgE production, while IL-5 affects eosinophil production, activation, and survival, and eosinophils are activated by IgE. Therefore, IL-4, IL-5, and eosinophil activity are associated with increased IgE.

[0057] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0058] In one embodiment, the allergic disease can be an IgE (Immunoglobulin E) mediated or non-IgE mediated disease or condition, or a condition associated with an allergic disease.

[0059] The term "treat" as used herein may mean that inflammation or a bacterial infection is cured in a short time compared to natural healing. The treatment may include improvement and / or alleviation of inflammation or a bacterial infection. Furthermore, the treatment may mean the cure and / or recovery of symptoms resulting from inflammation or a bacterial infection.

[0060] 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.

[0061] 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, 10 5 ~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 consideration. The frequency of administration can be once a month, once every two weeks, once a week, once a day, twice a day, or three times a day. Specifically, it can be administered once or twice or more times within the range of clinically acceptable side effects, and it can be administered 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, intradermal, intravenous, intraarterial, intramuscular, intraperitoneal, intradural, intra-articular, intrasynovial, intrasternal, or intralesional), rectal, topical (including transdermal), inhalation, and injection, or insertion of an implantable device or material. Target animals for treatment in one embodiment include humans and other mammalian species, specifically 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, once to three times daily.

[0062] Another aspect is to provide a health functional food composition for preventing or ameliorating allergic diseases, which contains a lactiplantibacillus plantarum strain, a lysate derived from said strain, a culture solution, or a mixture thereof as an active ingredient.

[0063] The "strain", "antiallergic activity", and "allergic disease" are as defined above.

[0064] In one embodiment, the health functional food composition may have anti-allergic activity.

[0065] In one embodiment, the anti-allergic activity may be suppression of Th2-mediated cytokine production, suppression of mast cell degranulation activity, suppression of serum IgE levels, or suppression of serum MCPT-1 levels. Specifically, the suppression of mast cell degranulation activity may be suppression of β-hexosaminidase secretion in activated mast cells.

[0066] In one embodiment, the Th2-mediated cytokine may be at least one selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

[0067] In one embodiment, the health functional food composition may reduce serum IgE levels or reduce the binding of IgE to surface receptors on mast cells, and the strain may suppress serum MCPT-1 levels.

[0068] The functional health food composition is preferably prepared in any dosage form selected from powder, granules, rings, tablets, capsules, candies, syrups, and beverages, but is not limited thereto. The functional health food composition of the present invention can be prepared by adding the active ingredient directly or by mixing it with other foods or food ingredients, and can be prepared appropriately according to conventional methods. Examples of foods to which the keigai extract can be added include caramel, meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, dairy products including infant formula, milk, yogurt, cheese, fermented milk, milk powder, various soups, drinking water, tea, supplementary drinks, alcoholic beverages, and vitamin complexes, including all health functional foods in the conventional sense. In other words, the type of food is not particularly limited. The health functional food composition may contain various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectinic 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 beverages, etc. Furthermore, fruit pulp may be included for the production of natural fruit juices and vegetable beverages. The above ingredients may be used independently or in combination.

[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 the degranulation activity of mast cells, or suppressing serum IgE or MCPT-1 levels.

[0071] In one embodiment, the allergic disease may be selected from the group consisting of edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, pruritus, anaphylactic shock, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0072] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0073] 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 fruit pulp for the production of natural fruit juice, fruit juice drinks, vegetable drinks, or combinations thereof.

[0074] Another aspect is to provide a pharmaceutical composition for treating or preventing allergic diseases, comprising a lactiplantibacillus plantarum strain, a lysate or culture medium derived from said strain, or a mixture thereof, as an active ingredient.

[0075] The "strain", "antiallergic activity" and "allergic disease" are as defined above.

[0076] In one embodiment, the pharmaceutical composition may have anti-allergic activity.

[0077] In one embodiment, the anti-allergic activity may be suppression of Th2-mediated cytokine production, suppression of mast cell degranulation activity, suppression of serum IgE levels, or suppression of serum MCPT-1 levels. Specifically, the suppression of mast cell degranulation activity may be suppression of β-hexosaminidase secretion in activated mast cells.

[0078] In one embodiment, the Th2-mediated cytokine may be at least one selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

[0079] In one embodiment, the pharmaceutical composition may reduce serum IgE levels or reduce the binding of IgE to mast cell surface receptors, and the bacterial strain may suppress serum MCPT-1 levels.

[0080] In one embodiment, the allergic disease may be selected from the group consisting of edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, pruritus, anaphylactic shock, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0081] In one embodiment, the allergic condition may include an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

[0082] 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.

[0083] The pharmaceutical composition can be formulated into an oral or parenteral dosage form. Specifically, it can be formulated for oral, rectal, intravenous, intravascular, intratumoral, subcutaneous, intradermal, or intraperitoneal delivery. More specifically, the oral dosage form can be granules, powders, liquids, tablets, capsules, dry syrups, or combinations thereof, and the parenteral dosage form can be an injection.

[0084] Another aspect is to provide a cosmetic composition 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.

[0085] The "strain", "antiallergic activity" and "allergic disease" are as defined above.

[0086] In one embodiment, the cosmetic composition may have anti-allergic activity.

[0087] 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.

[0088] The ingredients contained in the cosmetic composition may include ingredients commonly used in cosmetic compositions in addition to the composition as active ingredients, and may include conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0089] 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.

[0090] In one embodiment, the individual's condition can be a disease or condition mediated by IgE (Immunoglobulin E) or a condition that is not mediated by IgE, or a condition associated with an allergic disorder.

[0091] As used herein, the term "administration" refers to the physical introduction of a composition into a subject using any of a variety of methods and delivery systems known to those skilled in the relevant art. The administration can be by any method known in the art. The administration can be directly administered to an individual by any means, for example, via intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous administration. The administration can be systemic or local.

[0092] 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.

[0093] 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]

[0094] According to one embodiment, the novel strain and its derived lysate, culture medium, extract of the culture medium, or mixtures thereof can be effectively used for the prevention, amelioration, or treatment of allergic diseases. [Brief explanation of the drawings]

[0095] [Figure 1] FIG. 1 is a genetic map of the GBCC_F0070 strain. [Figure 2] FIG. 2 is a graph showing the degranulation inhibitory effect of GBCC_F0070 culture supernatant on RBL-2H3 mast cells. [Figure 3] FIG. 3 is a graph showing the ear edema-inhibitory effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis. [Figure 4] FIG. 4 is a graph showing the serum IgE-reducing effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis. [Figure 5] FIG. 5 is a graph showing the serum MCPT-1 reducing effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis. [Figure 6] FIG. 6 is a graph showing the inhibitory effect of the L. plantarum GBCC_F0070 strain on IgE-mediated allergic reactions in a manual cutaneous anaphylaxis animal model. [Figure 7] FIG. 7 is a graph showing the allergic symptom-improving effect in the group administered with the L. plantarum GBCC_F0070 strain in an animal food allergy model. [Figure 8] FIG. 8 is a graph showing the effect of reducing serum total IgE in the group administered with the L. plantarum GBCC_F0070 strain in an animal food allergy model. [Figure 9] FIG. 9 is a graph showing the effect of reducing inflammatory cells in bronchoalveolar lavage fluid (BALF) in the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model. [Figure 10] FIG. 10 is a graph showing the serum total IgE reducing effect in the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model. [Figure 11] FIG. 11 is a graph showing the Th2 cytokine suppression effect in serum of the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model. [Figure 12] FIG. 12 is a graph showing the suppressive effect of Th2 cytokines (IL-4, IL-5) in the nasal mucosa of the group administered with the L. plantarum GBCC_F0070 strain in an animal model of ovalbumin-induced rhinitis. [Figure 13]FIG. 13 is a graph showing the effect of reducing serum total IgE and ovalbumin-specific IgE in the group administered with the L. plantarum GBCC_F0070 strain in an animal model of ovalbumin-induced rhinitis. DETAILED DESCRIPTION OF THE INVENTION

[0096] 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.

[0097] Example 1 Isolation and identification of Lactiplantibacillus plantarum strain

[0098] 1.1. Strain isolation The Lactiplantibacillus plantarum strain of the present invention was isolated from home-made young radish kimchi. Young radish kimchi 23 days after pickling was serially diluted 1:10 in PBS buffer and plated onto MRS (deMan, Rogosa, Sharpe) plates, a lactic acid bacteria culture medium. After incubation in an anaerobic chamber at 37°C for 2 days, bacterial colonies were observed, and single colonies were subcultured onto new MRS plates for pure isolation. The identity of the pure isolates was confirmed by PCR of the 16S rRNA gene and Sanger blotting. For long-term storage of the strain, the strain was cultured in liquid medium, and upon reaching exponential growth phase, 30% glycerol was added at a 1:1 ratio and stored in a deep freezer at -70°C. Cultures after pure isolation and identification were also performed on MRS plates and liquid medium. After 48 hours of culture on MRS solid medium, large, smooth, round white colonies were formed.

[0099] Using the above method, the Lactiplantibacillus plantarum GBCC_F0070 (hereinafter also referred to as "GBCC_F0070") strain was finally selected.

[0100] 1.2. Molecular identification of selected strains The identification of the Bacillus plantarum GBCC_F0070 strain was based on sequence similarity between the 16S rRNA gene and the recA gene. DNA was extracted from the bacterial culture using the FastDNA® SPIN Kit for soil (MP Bio, 6560200), and PCR was performed using the primer sequences shown in Table 1.

[0101] [Table 1] The sequence produced using the 27F and 1492R primers to PCR amplify the bacterial 16S rRNA gene is shown in SEQ ID NO: 1. GBCC_F0070 showed 98.5% or higher sequence similarity to 13 Lactiplantibacillus sp. species, including the reference strain of Lactiplantibacillus plantarum, in terms of 16S rRNA sequence, confirming that GBCC_F0070 corresponds to one of these 13 strains (Table 2).

[0102] [Table 2] Before the strain name was changed to Lactiplantibacillus plantarum, it was named Lactobacillus plantarum. In addition to this, a more accurate identification was performed based on sequence similarity of the recA gene, which is widely used as a bacterial taxonomic biomarker, in addition to the 16S rRNA gene. As a result, GBCC_F0070 was confirmed to be the strain most closely related to the species Lactiplantibacillus plantarum (Table 3).

[0103] [Table 3] The recA gene sequence of GBCC_F0070 is shown in SEQ ID NO:2.

[0104] Example 2. Genomic and comparative genetic analysis of Lactiplantibacillus plantarum strains

[0105] To identify and characterize the genome-based species of the GBCC_F0070 strain, we utilized next-generation sequencing (NGS) and bioinformatics techniques to fully sequence the genome of the strain and infer the functions of the genes contained in the genome.

[0106] Genome sequencing confirmed that GBCC_F0070 possesses one prototype chromosome of 3,259,176 bp and six plasmids, and that GBCC_F0070 contains 3,140 CDSs, 16 rRNA genes, and 66 tRNA genes (Fig. 1, Table 4).

[0107] [Table 4] The average nucleotide identity (ANI) of the genome of the GBCC_F0070 strain and the genome of the same species strain published in GenBank was calculated using the Jspecies program. The sequence similarity with the closest strain was 99.16%, indicating that GBCC_F0070 is a novel strain that has not been reported before (Table 5).

[0108] [Table 5] Therefore, the inventors named the GBCC_F0070 strain "Lactiplantibacillus plantarum GBCC_F0070" (deposit number: KCTC 15051BP) and deposited it with the Korean Collection for Type Cultures (KCTC), Korea Institute of Bioscience and Biotechnology, on August 16, 2022.

[0109] Experimental Example 1. Degranulation inhibitory effect of L. plantarum GBCC_F0070 culture supernatant on RBL-2H3 mast cells

[0110] To demonstrate therapeutic activity against IgE-mediated allergies, we examined the inhibitory effect on mast cell degranulation. Culture supernatants of various L. plantarum strains, including the GBCC_F0070 strain, were treated with L. plantarum strains, which were activated by allergic reactions, and the inhibitory effect on degranulation was screened by measuring the secretion of β-hexosaminidase. β-hexosaminidase is stored within the secretory granules of mast cells and secretes histamine extracellularly during allergic reactions, so it can be used as an indicator of mast cell degranulation.

[0111] RBL-2H3 cells were suspended in MEM medium containing 15% FBS in a 24-well plate, and then 1.5 × 10 cells were added per well. 5Cells were 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, precipitating the strains by centrifugation, 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 2.

[0112] FIG. 2 is a graph showing the degranulation inhibitory effect of GBCC_F0070 culture supernatant on RBL-2H3 mast cells.

[0113] As shown in 2, after screening the culture supernatants of 26 different L. plantarum strains, we confirmed that the culture supernatant of the L. plantarum GBCC_F0070 strain inhibited mast cell degranulation by approximately 61%, making it the most effective in suppressing IgE-mediated allergic reactions.

[0114] Experimental Example 2: Inhibitory effect of ear edema in the L. plantarum GBCC_F0070 strain-administered group in an MC903-induced atopic dermatitis animal model

[0115] The inhibitory effect of L. plantarum GBCC_F0070 on ear edema was confirmed in an animal model of MC903-induced atopic dermatitis.

[0116] Six-week-old BALB / c mice were obtained. After a one-week purification period, freeze-dried L. plantarum GBCC_F0070 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. 9 After 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.

[0117] FIG. 3 is a graph showing the ear edema-inhibitory effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis.

[0118] As shown in Figure 3, in the MC903-induced atopic dermatitis model, the experimental group (MC903+GBCC_F0070) administered with the L. plantarum GBCC_F0070 strain showed an approximately 34% inhibitory effect on ear edema compared to the control group (MC903).

[0119] Experimental Example 3: Reduction of serum total IgE in the L. plantarum GBCC_F0070 strain-administered group in an MC903-induced atopic dermatitis animal model

[0120] 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 in Experimental Example 3.

[0121] 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 1:1000 were added and incubated for 2 hours at room temperature. After washing four times, the detection antibody was added and incubated 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. Stop solution was added to each well to stop the color reaction, and the absorbance was measured at 450 nm using an ELISA reader. The results are shown in Figure 4.

[0122] FIG. 4 is a graph showing the serum IgE-reducing effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis.

[0123] 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+GBCC_F0070) administered with the L. plantarum GBCC_F0070 strain showed a reduction in serum total IgE concentration of approximately 62%.

[0124] Experimental Example 4: Serum MCPT-1 reduction effect in the L. plantarum GBCC_F0070 strain-administered group in an MC903-induced atopic dermatitis animal model

[0125] To confirm the serum mast cell protease-1 (MCPT-1) concentration in the MC903-induced atopic dermatitis animal model in Experimental Example 2, a mouse MCPT-1 (mMCP-1) ELISA kit (Invitrogen #88-7503) was used to measure it.

[0126] 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.

[0127] FIG. 5 is a graph showing the serum MCPT-1 reducing effect of GBCC_F0070 in a mouse model of MC903-induced atopic dermatitis.

[0128] 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+GBCC_F0070) administered with the L. plantarum GBCC_F0070 strain showed a reduction in serum MCPT-1 concentration of approximately 57%.

[0129] Experimental Example 5. Inhibitory effect of L. plantarum GBCC_F0070 on IgE-mediated allergic reactions in an animal model of passive cutaneous anaphylaxis

[0130] The inhibitory effect of L. plantarum GBCC_F0070 strain on the allergic reaction was confirmed in an animal model of passive cutaneous anaphylaxis (PCA) induced by anti-DNP IgE.

[0131] Four-week-old BALB / c mice were obtained, and after a one-week purification period, freeze-dried L. plantarum GBCC_F0070 strain was suspended in D-PBS. The mice were inoculated with 1 × 10 L. plantarum GBCC_F0070 at a concentration 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 100 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, and the results are shown in Figure 6.

[0132] FIG. 6 is a graph showing the inhibitory effect of the L. plantarum GBCC_F0070 strain on IgE-mediated allergic reactions in a manual cutaneous anaphylaxis animal model.

[0133] As shown in Figure 6, the anti-allergic effect was confirmed by suppressing the allergic reaction by approximately 24% in the experimental group (Ag+GBCC_F0070) administered with the L. plantarum GBCC_F0070 strain compared to the negative control group (Ag) in a passive skin sensitization allergic reaction model using anti-DNP IgE.

[0134] Experimental Example 6: Improvement of allergic symptoms in a food allergy animal model by administering L. plantarum GBCC_F0070 strain

[0135] The effect of ovalbumin (OVA) on improving allergic symptoms caused by L. plantarum GBCC_F0070 strain was investigated in an animal model of food allergy.

[0136] Five-week-old BALB / c mice were obtained and, after a one-week purification period, 50 μg of OVA and 1 mg of aluminum hydroxide were mixed and intraperitoneally injected twice at two-week intervals to induce a food allergic reaction. Two weeks after the final sensitization, 50 mg of egg albumin was orally administered five times at two-day intervals to induce an allergic reaction. Furthermore, to confirm the therapeutic effect of allergies, lyophilized L. plantarum GBCC_F0070 strain was suspended in D-PBS and administered at 2 × 10 per mouse from the time of egg albumin sensitization until just before the end of the test. 9 The fecal morphology due to food allergy was visually observed, and diarrhea symptoms were scored. The results are shown in Figure 7 and Table 6.

[0137] [Table 6] Figure 7 is a graph showing the allergic symptom improvement effect in the group administered with the L. plantarum GBCC_F0070 strain in a food allergy animal model. As shown in Figure 7, the diarrhea symptom score was significantly increased in the control group (OVA) in which food allergy was induced, compared to the normal group. In the egg albumin-induced food allergy reaction model, the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed an approximately 26% improvement in diarrhea symptoms compared to the control group.

[0138] Experimental Example 7: Effect of L. plantarum GBCC_F0070 strain administration on serum total IgE levels in a food allergy animal model

[0139] To confirm the serum total IgE concentration in the egg albumin-induced food allergy animal model, it was measured using a mouse IgE ELISA kit (Biolegend #432404).

[0140] The capture antibody diluted in coating buffer was applied to a microwell plate overnight at 4°C, then 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 1:1000 were added and incubated at room temperature for 2 hours. After washing four times, the detection antibody was added and incubated at room temperature for 1 hour. After washing four times, avidin-HRP was added and incubated at room temperature for 30 minutes. After washing five times, TMB substrate solution was added and incubated at room temperature for 20 minutes in the dark. The color reaction was stopped by adding reaction stop solution to each well, and the absorbance was measured at 450 nm using an ELISA reader. The results are shown in Figure 8.

[0141] FIG. 8 is a graph showing the effect of reducing serum total IgE in the group administered with the L. plantarum GBCC_F0070 strain in an animal food allergy model.

[0142] As shown in Figure 8, the control group (OVA) showed a rapid increase in serum total IgE concentration compared to the normal group, but the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed a reduction in serum total IgE concentration of approximately 21%.

[0143] Experimental Example 8. Effect of L. plantarum GBCC_F0070 strain administration on the reduction of inflammatory cells in bronchoalveolar lavage fluid (BALF) in an animal model of egg albumin-induced asthma

[0144] The asthma-relieving effect of L. plantarum GBCC_F0070 strain was confirmed in an animal model in which bronchial asthma was induced with ovalbumin (OVA).

[0145] Five-week-old BALB / c mice were obtained and, after a one-week purification period, were sensitized twice, intraperitoneally, with a mixture of 50 μg of OVA and 1 mg of ammonium hydroxide, at two-week intervals to create an asthma mouse model. One week after the final sensitization, from the 21st to the 25th, mice were anesthetized and 150 μg of ovalbumin was instilled intranasally and inhaled into the lungs to induce asthma. 24 hours after the final challenge, the mice were anesthetized, the bronchi were opened, and bronchoalveolar lavage fluid was collected and the total cell count was calculated. The results are shown in Figure 9.

[0146] FIG. 9 is a graph showing the effect of reducing inflammatory cells in bronchoalveolar lavage fluid (BALF) in the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model.

[0147] As shown in Figure 9, the number of inflammatory cells in the bronchoalveolar lavage fluid of the asthma-induced group was significantly increased compared to the normal group, and the number of inflammatory cells in the bronchoalveolar lavage fluid of the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) was reduced by approximately 64% compared to the control group (OVA).

[0148] Experimental Example 9. Reduction of serum total IgE and egg albumin-specific IgE in the L. plantarum GBCC_F0070 strain-administered group in an egg albumin-induced asthma animal model

[0149] To confirm serum total IgE and ovalbumin (OVA)-specific IgE concentrations in an ovalbumin-induced asthma animal model, measurements were performed using a mouse IgE ELISA kit (Biolegend #432404) and a mouse OVA-specific IgE ELISA kit (Biolegend #439807).

[0150] For total serum IgE, a microwell plate was coated with a capture antibody diluted in coating buffer overnight at 4°C, washed four times with wash buffer, and then blocked with assay diluent for 1 hour at room temperature. The plate was washed four times, and then a standard sample and serum samples diluted 1:500 (total IgE) and 1:2 (ovalbumin-specific IgE) were added and incubated for 2 hours 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, protected from light. 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 10.

[0151] FIG. 10 is a graph showing the serum total IgE reducing effect in the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model.

[0152] As shown in Figure 10, the control group (OVA) showed a rapid increase in serum total IgE concentration compared to the normal group, but the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed a reduction in serum total IgE concentration of approximately 37%.

[0153] Experimental Example 10. Suppression of serum Th2 cytokines in the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model

[0154] To confirm serum IL-5 concentrations in the ovalbumin-induced asthma animal model, they were measured using a mouse IL-5 ELISA kit (Biolegend #431204).

[0155] The capture antibody diluted in coating buffer was applied to a microwell plate and incubated overnight at 4°C. The plate was then 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 1:5 were added and incubated for 2 hours at room temperature. After washing four times, the detection antibody was added and incubated 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, protected from light. 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 11.

[0156] FIG. 11 is a graph showing the Th2 cytokine suppression effect in serum of the L. plantarum GBCC_F0070 strain-administered group in an ovalbumin-induced asthma animal model.

[0157] As shown in Figure 11, the control group (OVA) showed a rapid increase in serum IL-5 concentration compared to the normal group, but the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed a statistically significant decrease of approximately 88% in serum IL-5 production compared to the control group.

[0158] This confirms that the L. plantarum GBCC_F0070 strain has an inhibitory effect on serum Th2 cytokines, meaning that it has therapeutic activity not only against IgE-mediated allergies but also against non-IgE-mediated allergies.

[0159] Experimental Example 11. Inhibitory effect of L. plantarum GBCC_F0070 on Th2 cytokines (IL-4, IL-5, IL-13) in the nasal mucosa in an ovalbumin-induced rhinitis animal model

[0160] The effect of L. plantarum GBCC_F0070 strain on alleviating rhinitis was investigated in an animal model in which rhinitis was induced with egg albumin (OVA).

[0161] Five-week-old BALB / c mice were obtained and, after a one-week purification period, were sensitized three times, intraperitoneally, with a mixture of 50 μg of OVA and 2 mg of ammonium hydroxide at one-week intervals to create a mouse model of rhinitis. One week after the final sensitization, rhinitis was induced by intranasal instillation of 100 μg of ovalbumin from the 21st to the 26th of each day without anesthesia to prevent antigen aspiration into the lungs. 24 hours after the final challenge, the nasal mucosa was removed and disrupted to isolate RNA, which was then synthesized into cDNA. Expression levels of mRNA for Th2 cytokines (IL-4, IL-5, and IL-13) were measured using a QuantStudio 3 Real-Time PCR Instrument. The results are shown in Figure 12.

[0162] FIG. 12 is a graph showing the suppressive effect of Th2 cytokines (IL-4, IL-5) in the nasal mucosa of the group administered with the L. plantarum GBCC_F0070 strain in an animal model of ovalbumin-induced rhinitis.

[0163] As shown in Figure 12, Th2 cytokines in the nasal mucosa of the rhinitis-induced group were significantly increased compared to the normal group. Compared to the control group (OVA), the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed a statistically significant suppression effect of approximately 25% on IL-4 mRNA expression, and a 21% decrease in IL-5 mRNA expression was confirmed.

[0164] This confirms that the L. plantarum GBCC_F0070 strain has an inhibitory effect on serum Th2 cytokines, meaning that it has therapeutic activity not only against IgE-mediated allergies but also against non-IgE-mediated allergies.

[0165] Experimental Example 12. Reduction of serum total IgE and egg albumin-specific IgE in the L. plantarum GBCC_F0070 strain-administered group in an egg albumin-induced rhinitis animal model

[0166] To confirm the serum total IgE and ovalbumin (OVA)-specific IgE concentrations in the ovalbumin-induced rhinitis animal model, measurements were performed using a mouse IgE ELISA kit (Biolegend #432404) and a mouse OVA-specific IgE ELISA kit (Biolegend #439807).

[0167] Total serum IgE was measured by incubating the capture antibody diluted in coating buffer on a microwell plate overnight at 4°C, washing four times with wash buffer, and blocking with assay diluent for 1 hour at room temperature. The plate was washed four times, and then a standard sample and serum samples diluted 1:500 (total IgE) and 1:2 (ovalbumin-specific IgE) were added and incubated for 2 hours 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, protected from light. 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 13.

[0168] FIG. 13 is a graph showing the effect of reducing serum total IgE and ovalbumin-specific IgE in the group administered with the L. plantarum GBCC_F0070 strain in an animal model of ovalbumin-induced rhinitis.

[0169] As shown in Figure 13, the control group (OVA) showed a rapid increase in serum total IgE levels compared to the normal group, but the experimental group administered with the L. plantarum GBCC_F0070 strain (OVA+GBCC_F0070) showed a reduction in serum total IgE levels of approximately 14%.In addition, the experimental group administered with the L. plantarum GBCC_F0070 strain showed a reduction in serum ovalbumin-specific IgE levels of approximately 22% compared to the control group.

[0170] These results indicate that the L. plantarum GBCC_F0070 strain can be useful for preventing, ameliorating, or treating atopic dermatitis or allergic diseases.

[0171] 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 and experimental examples are illustrative in all respects and are not limiting. [Accession number]

[0172] Contracted organization name: Korea Institute of Bioscience and Biotechnology Accession number: KCTC15051BP Entrustment date: August 16, 2022

Claims

1. A composition for suppressing the production of Th2 (helper T2 cell)-mediated cytokines or alleviating allergic reactions, comprising a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus as an active ingredient.

2. Lactiplantibacillus plantarum GBCC_F0070 strain belonging to the genus Lactiplantibacillus sp., deposited under accession number KCTC15051BP.

3. The strain of claim 2 , wherein the strain comprises the 16S rRNA sequence of SEQ ID NO:

1.

4. The strain of claim 2 , wherein the strain comprises the recA gene sequence of SEQ ID NO:

2.

5. The strain of claim 2 , wherein the strain comprises a naturally occurring mutation of a Lactiprantibacillus plantarum strain.

6. 3. The bacterial strain according to claim 2, wherein the bacterial strain is a live bacterium, a killed bacterium, an attenuated bacterium, or a cytoplasmic fraction obtained by disrupting the bacterial strain.

7. The strain of claim 2, wherein the strain comprises the following characteristics: - suppression of Th2-mediated cytokine production, - Inhibition of mast cell degranulation activity, - suppression of serum IgE levels, or - Suppression of serum MCPT-1 levels.

8. The strain of claim 7, wherein the Th2-mediated cytokine is at least one selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

9. A composition comprising a lysate, a culture medium, or an extract of a culture medium derived from the strain of claim 2.

10. 10. The composition of claim 9, wherein the composition is for preventing or treating an IgE-mediated disease or condition.

11. 10. The composition of claim 9, wherein the composition reduces serum IgE levels or reduces the binding of IgE to surface receptors on mast cells.

12. 10. The composition of claim 9, wherein the composition further comprises an additive selected from the group consisting of preservatives, dyes, emulsifiers, sweeteners, stabilizers, flavor enhancers, flavoring agents, and acidulants.

13. 10. The composition of claim 9, which is at least one formulation selected from the group consisting of a solution, an emulsion, a viscous mixture, a powder, a granule, a tablet, and a capsule.

14. A functional health food composition for improving or preventing allergic diseases, comprising, as an active ingredient, Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, a disruption liquid derived from said strain, a culture liquid, or a mixture thereof.

15. The health functional food composition according to claim 14, wherein the improvement of the allergic disease comprises: - suppression of Th2-mediated cytokine production, - Inhibition of mast cell degranulation activity, - suppression of serum IgE levels, or - Suppression of serum MCPT-1 levels.

16. The strain of claim 15, wherein the Th2-mediated cytokine is at least one selected from the group consisting of IL-4, IL-5, IL-6, IL-10, and IL-13.

17. The health functional food composition according to claim 14, 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.

18. The health functional food composition according to claim 14, wherein the allergic disease is selected from the group consisting of edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, pruritus, anaphylactic shock, insect allergy, food allergy, drug allergy, and respiratory allergy.

19. The health functional food composition according to claim 14, wherein the allergic disease includes allergic reactions caused by house dust, fungi, mites, or animal hair, skin, or feces.

20. A pharmaceutical composition for treating or preventing an allergic disease, comprising the Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, a lysate or culture solution derived from said strain, or a mixture thereof, as an active ingredient.

21. The pharmaceutical composition of claim 20, wherein the composition has mast cell degranulation inhibitory activity.

22. 21. The pharmaceutical composition of claim 20, wherein the composition i) suppresses serum IgE expression, or ii) suppresses serum MCPT-1 levels.

23. 21. The pharmaceutical composition according to claim 20, wherein the allergic disease is selected from the group consisting of edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, pruritus, insect allergy, food allergy, drug allergy, and respiratory allergy.

24. 21. The pharmaceutical composition of claim 20, wherein the allergic disease comprises an allergic reaction caused by house dust, fungi, mites, or animal hair, skin, or feces.

25. 21. The pharmaceutical composition of claim 20, wherein the composition can be administered orally or parenterally.

26. 26. The pharmaceutical composition of claim 25, wherein the oral or parenteral administration is administered once a month, once every two weeks, once a week, once a day, twice a day, or three times a day.

27. 27. The pharmaceutical composition of claim 26, wherein the parenteral administration is subcutaneous, intradermal, intravenous, intraarterial, intramuscular, intraperitoneal, intradural, intra-articular, intrasynovial, intrasternal, intralesional or intrarectal administration.

28. 21. The pharmaceutical composition of claim 20, wherein the composition is formulated for oral, rectal, intravenous, intravascular, intratumoral, subcutaneous, intradermal, or intraperitoneal delivery.

29. A cosmetic composition for improving or preventing skin allergies, comprising the Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, a lysate or culture solution derived from said strain, or a mixture thereof.

30. 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 GBCC_F0070 strain deposited under accession number KCTC15051BP, a culture of said strain, a disrupted product of said strain, or a composition containing a mixture thereof.

31. Use of a composition comprising the Lactiplantibacillus plantarum GBCC_F0070 strain deposited under accession number KCTC15051BP, 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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