Use of Lactobacillus plantarum strains for ameliorating skin conditions caused by allergy

Abstract

The invention relates to application of a lactobacillus plantarum strain in improving the skin state caused by allergy, and the strain and derivative lysate, culture solution, extract of the culture solution or mixture of the strain have the effect of being effectively used for preventing, improving or treating allergic diseases according to a scheme.

Description

Technical Field

[0001] The present invention relates to the use of a Lactobacillus plantarum strain in improving skin conditions caused by allergies. Background Art

[0002] Allergy or hypersensitivity is a compound word formed by combining "allos" meaning "change" and "ergo" meaning "action" in Greek, and it means an abnormal reaction of an organism. It can be inferred from the etymology that allergy refers to a hypersensitivity reaction in a specific population caused by an allergen, which is a substance that does not cause any reaction in ordinary people, due to abnormal innate or acquired immune functions, such as urticaria, itching, runny nose, and cough. Typical allergens include pollen, drugs, plant fibers, bacteria, molds, dust mites, foods (ovalbumin, milk protein, peanuts), dyes, and chemical substances, etc. In recent years, due to the increase in indoor living, the increase in allergen induction by new material chemical substances, environmental pollution, immune function decline caused by stress, and changes in eating habits, these allergic diseases have shown an upward trend globally. Typical allergic diseases include atopic dermatitis, bronchial asthma, allergic rhinitis, allergic keratitis, skin urticaria, etc., and it is known that mast cells are important mediator cells in these allergic diseases.

[0003] When an allergen binds to IgE bound to the high-affinity IgE receptor (FcεRI) on the surface of these mast cells, an allergic reaction is induced by activating the intracellular signaling pathway. Specifically, when an allergen invades the body, T-helper cells and B cells are sequentially activated to generate IgE. The allergen invading the body is recognized by an antigen-presenting cell, which differentiates Th0 (T helper cell type 0) into Th2 cells through the presentation of the allergen. These differentiated Th2 cells secrete cytokines such as IL-4, IL-5, and IL-13, which can not only promote the development of eosinophils in the bone marrow and direct eosinophils to the inflamed tissue, but also act on B cells to induce the production of IgE. The IgE thus produced binds to the high-affinity IgE receptor (FcRIα) on the mast cell membrane. When allergens such as mites and pollen bind to it and form crosslinks, the receptor (FcRIα) will aggregate. Mast cells are activated through the crosslinking and aggregation of the receptor (FcRIα), thereby increasing the concentration of calcium ions (Ca++) in the cytoplasm. The increase in calcium ion concentration acts on various intracellular regulatory factors such as cholesterol in the cell membrane or the cytoskeleton system, causing the granules to move towards the cell membrane, and at the same time triggering granule vacuolization, inter-granular fusion, and fusion of granules with the cell membrane, resulting in degranulation. During degranulation, histamine, prostaglandin, leukotriene, serotonin, etc. are secreted, thereby causing vasodilation, increased vascular permeability, bronchial smooth muscle contraction, etc. The degranulation of mast cells is accompanied by the secretion of inflammatory cytokines such as IL-1β, IL-4, IL-5, IL-6, and TNF-α, and neutrophils, eosinophils, macrophages, Th2 cells, basophils, etc. infiltrate into the corresponding tissues, thereby inducing an allergic inflammatory reaction.

[0004] Therefore, substances that inhibit the activation of mast cells, inhibit degranulation, or inhibit the production of inflammatory cytokines may become effective anti-allergy therapeutic drugs. Currently, drugs used for anti-allergy include antihistamines (alkylamines, dexloratadine, etc.), mast cell stabilizers (cromoglycate, etc.), leukotriene receptor antagonists (montelukast, etc.), anti-IgE antibodies (Omalizumab), etc. However, the efficacy of such drugs is mostly temporary, and continuous high-concentration treatment can cause side effects. Therefore, their application is mostly limited in the treatment of allergic diseases with chronic immune diseases.

[0005] Therefore, there is still a need to develop new substances with the effects of preventing and improving allergic diseases, having fewer side effects, and a long-lasting therapeutic effect. Summary of the Invention Technical Problem

[0006] In one aspect, there is provided a Lactiplantibacillus plantarum strain having anti-allergic activity, wherein the anti-allergic activity is to reduce the concentration of serum IgE or inhibit the degranulation activity of mast cells.

[0007] In another aspect, there is provided a Lactiplantibacillus plantarum HD-02 strain belonging to the genus Lactiplantibacillus, which is deposited under the accession number KACC81228BP.

[0008] In still another aspect, there is provided a lysate, a culture solution, or an extract of the culture solution derived from the strain.

[0009] In still another aspect, there is provided a food composition, which comprises a Lactiplantibacillus strain, a lysate, a culture solution derived from the strain, or a mixture thereof as an active ingredient.

[0010] In still another aspect, there is provided a health functional food for preventing or improving allergic diseases, which comprises a Lactiplantibacillus strain, a lysate, a culture solution derived from the strain, or a mixture thereof as an active ingredient.

[0011] In still another aspect, there is provided a pharmaceutical composition for treating or preventing allergic diseases, which comprises a Lactiplantibacillus strain, a lysate, a culture solution derived from the strain, or a mixture thereof as an active ingredient.

[0012] In still another aspect, there is provided a cosmetic composition for preventing or improving allergic diseases, which comprises a Lactiplantibacillus strain, a lysate, a culture solution derived from the strain, or a mixture thereof as an active ingredient.

[0013] In still another aspect, there is provided a feed composition for preventing or improving allergic diseases, which comprises a Lactiplantibacillus strain, a lysate, a culture solution derived from the strain, or a mixture thereof as an active ingredient.

[0014] In yet another aspect, there is provided a method for preventing or treating allergic diseases, the method comprising the step of administering to a subject in need thereof a composition comprising an effective amount of Lactiplantibacillus plantarum strain HD-02 deposited under accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof.

[0015] In yet another aspect, there is provided the use of a composition in the preparation of a pharmaceutical preparation or a health functional food preparation for preventing or treating allergic diseases, wherein the composition comprises Lactiplantibacillus plantarum strain HD-02 deposited under accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof. Technical solution

[0016] In one aspect, there is provided a Lactiplantibacillus plantarum strain belonging to the genus Lactiplantibacillus.

[0017] Lactiplantibacillus is a genus of aerobic or facultatively anaerobic Gram-positive bacilli that are widely distributed in nature. Microorganisms belonging to the genus Lactiplantibacillus include Lactiplantibacillus plantarum, Lactiplantibacillus fermentum, etc. The present inventors conducted research to develop new strains with excellent anti-allergic effects and screened Lactiplantibacillus plantarum HD-02 as a candidate anti-allergic strain. The strain was deposited at the Korean Agricultural Genebank on August 26, 2022, under the accession number KACC81228BP. The strain is equivalent to a probiotic strain, is harmless to the human body, and has no side effects when used.

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

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

[0020] In a specific example, the strain may be a strain having 16S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1, or a strain having 16S rRNA with a nucleotide sequence having a homology of more than 97% with its nucleotide sequence. Specifically, it has a homology of at least 93%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% with the nucleotide sequence consisting of SEQ ID NO: 1 in this specification.

[0021] In a specific example, the Lactobacillus plantarum strain may be a strain containing the recA gene sequence of SEQ ID NO: 2.

[0022] In a specific example, the strain may be a strain having the recA gene sequence containing SEQ ID NO: 2, or a recA with a gene sequence having a homology of more than 97% with its gene sequence. Specifically, it has a homology of at least 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.8%, 99.9% or 100% with the gene sequence consisting of SEQ ID NO: 2 in this specification.

[0023] In a specific example, the strain may be a naturally occurring mutant strain.

[0024] In a specific example, the strain may be a live bacterium, a dead bacterium or a cytoplasmic fraction obtained by disrupting the strain, and preferably may be a live bacterium. The strain may exist as a live or dead cell, or in a dried or lyophilized form.

[0025] In a specific example, the strain may have anti-allergic (hereinafter, allergy and allergic disease may be used interchangeably) activity.

[0026] In a specific example, the strain may have the activity of inhibiting the production of Th2-mediated cytokines, inhibiting the degranulation activity of mast cells, inhibiting the serum IgE level, or inhibiting the serum MCPT-1 level. Specifically, the inhibition of the mast cell degranulation activity may be the inhibition of the secretion of β-hexosaminidase in activated mast cells.

[0027] In a specific example, the strain may reduce the concentration of serum IgE or reduce the binding of IgE to the surface receptor of mast cells. In addition, the strain may inhibit the level of serum MCPT-1.

[0028] Therefore, the strain according to a specific example can be effectively used for the treatment of allergic diseases.

[0029] In another aspect, there is provided a lysate, a culture broth, an extract of the culture broth, or a mixture thereof, derived from a Lactiplantibacillus plantarum strain.

[0030] In the present specification, the term "culture broth" can be used interchangeably with "culture", "culture supernatant", "culture supernatant liquor", "conditioned culture medium" or "modified medium", and can refer to all culture media obtained after culturing the Lactobacillus strain in a culture medium capable of providing nutrients for the growth and survival of the Lactobacillus strain in vitro for a certain period of time, which contains the strain, its metabolites, additional nutrients, etc. The culture refers to the product obtained by culturing a probiotic strain in a known culture medium, and the product may or may not contain the strain itself. The culture medium can be selected from known liquid media or solid media, such as MRS liquid medium, GAM liquid medium, MRS agar medium, GAM agar medium, BL agar medium, but is not limited thereto.

[0031] The culture broth can include the culture broth itself obtained by culturing the strain, its concentrate, or freeze-dried product, or the culture supernatant liquor obtained by removing the strain from the culture broth, its concentrate or freeze-dried product.

[0032] In the present specification, the term "lysate" can be used interchangeably with "lysate", and refers to a solution or suspension of cells of a microorganism such as disrupted Lactiplantibacillus plantarum in an aqueous medium. The cell lysate includes, for example, macromolecules such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc., and / or micromolecules such as amino acids, sugars, fatty acids, etc., or components thereof. In addition, the lysate includes cell debris, the structure of which can be smooth or granular.

[0033] In the present specification, the term "extract of the culture broth" refers to the substance extracted from the culture broth or its concentrate, and can include the extract, the diluted or concentrated solution of the extract, the dried product obtained by drying the extract, or its crude or refined product, the fractionated product obtained by fractionating it.

[0034] The culture of the present invention can include a live bacteria culture (i.e., the product of culturing live bacteria in a culture medium), the culture after inactivating the live bacteria in the live bacteria culture, or a culture filtrate (i.e., the product obtained by removing dead or live bacteria from the culture). In addition, the components derived from the strain of the present application can include, for example, cell wall structural components obtained by disrupting the bacterial cells, or cytoplasmic components containing extracellular vesicles (EV), etc. In addition, the forms of lactic acid bacteria suitable for inclusion in pharmaceutical compositions, formulation methods, and separation methods of components derived from strains are well known to those skilled in the art.

[0035] In a specific example, the culture solution can be obtained by culturing Lactiplantibacillus plantarum in a suitable medium (for example, MRS plate medium) at any temperature higher than 10 °C or lower than 40 °C for a certain period of time, for example, 4 to 50 hours.

[0036] In a specific example, the medium and culture conditions for culturing the Lactiplantibacillus plantarum can be appropriately selected, modified and used by those skilled in the art.

[0037] In another aspect, a composition is provided, which comprises a lysate, a culture solution, an extract of the culture solution, or a mixture thereof, derived from a Lactiplantibacillus plantarum strain.

[0038] In a specific example, the composition further comprises additives selected from preservatives, dyes, emulsifiers, sweeteners, stabilizers, flavor enhancers, flavors and sour agents.

[0039] In a specific example, it can be more than one dosage form selected from solutions, emulsions, viscous mixtures, powders, granules, tablets and capsules.

[0040] In a specific example, the composition can have anti-allergic activity.

[0041] In a specific example, the anti-allergic activity can comprise the strain as an active ingredient, thereby inhibiting the production of Th2 (Helper T Cell Type 2)-mediated cytokines or alleviating allergic reactions. This can include the alleviation of immune allergic reactions, specifically, the improvement or alleviation of the skin condition caused by immune allergic reactions, for example, atopic dermatitis.

[0042] In a specific example, the anti-allergic activity can comprise inhibiting the degranulation of activated mast cells. Specifically, it can be inhibiting the secretion of β-hexosaminidase, histamine, prostaglandin and serine protease in activated mast cells.

[0043] In a specific example, the anti-allergic activity can comprise reducing the concentration of serum IgE or decreasing the binding of IgE to the surface receptors of mast cells. In addition, the anti-allergic activity can comprise inhibiting the level of serum MCPT-1.

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

[0045] According to one embodiment, the anti-allergic activity may include an activity of inhibiting ear swelling by 40% to 60%, an activity of reducing serum IgE concentration by 50% to 70%, or an activity of reducing the concentration of serum MCPT-1 by 50% to 70% when treating Lactobacillus plantarum strain HD-02 in an MC903-induced atopic dermatitis animal model.

[0046] According to one embodiment, the anti-allergic activity may include an activity of reducing serum IgE concentration by 50% to 70%, an activity of reducing transepidermal water loss by 30% to 60%, an activity of relieving itching symptoms by 40% to 70%, an activity of reducing the number of immune cells (specifically, B cells, T cells, Th1 cells, and Th2 cells) in lymph nodes (such as axillary lymph nodes) activated due to immune allergic reactions by 50% to 90%, an activity of inhibiting edema by 40% to 60%, or an activity of reducing the number of activated mast cells in inflamed tissues by 50% to 70% when treating Lactobacillus plantarum strain HD-02 in a house dust mite-induced atopic dermatitis animal model (severe atopic dermatitis model).

[0047] In a specific example, based on 100% of the allergic reaction symptoms of the negative control group that induces an allergic reaction or atopic dermatitis without applying the strain, Lactobacillus plantarum strain HD-02 may have the activity of reducing the symptoms of allergic reaction or atopic dermatitis to below 90%, below 80%, below 70%, below 67%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 67%, 20% to 90%, 20% to 80%, 20% to 70%, 20% to 67%, 30% to 90%, 30% to 80%, 30% to 70%, 30% to 67%, 40% to 90%, 40% to 80%, 40% to 70%, 40% to 67%, 50% to 90%, 50% to 80%, 50% to 70%, 50% to 67%, or 60% to 67% levels.

[0049] In yet another aspect, there is provided a use of Lactobacillus plantarum, a lysate, a culture solution, or an extract of the culture solution of the strain for improving, preventing, or treating diseases.

[0050] In a specific example, the use may include preventing, ameliorating or treating a disease or disorder mediated by IgE (Immunoglobulin E). Specifically, the disease mediated by IgE (Immunoglobulin E) may be an allergic disease.

[0051] In this specification, "allergic disease" refers to a hypersensitivity reaction of the human body to a specific substance, that is, a disease, disorder or abnormal condition induced by an overreaction of the immune system to an exogenous substance. The exogenous substance may be an antigen that causes an allergic disease, that is, an allergen. The allergy may refer to the release of inflammatory mediators such as histamine induced by an exogenous substance, thereby leading to a hypersensitivity reaction that induces the occurrence of a disease. The hypersensitivity reaction may be a type I hypersensitivity reaction, a type II hypersensitivity reaction, a type III hypersensitivity reaction or a type IV hypersensitivity reaction.

[0052] In a specific example, the allergic disease may include an allergic reaction caused by house dust, fungi, mites or the hair, skin or excrement of animals.

[0053] Non-limiting examples of the allergic disease may specifically be one or more diseases selected from edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic tympanitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, rash, dry eye, pruritus, anaphylactic shock, pollen allergy, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy and respiratory allergy, but are not limited thereto.

[0054] In this specification, the term "atopic dermatitis" is a skin eczema disease accompanied by chronic, recurrent and severe pruritus. When it becomes chronic, the epidermis thickens, and cells participating in various immune reactions infiltrate. The skin of patients with atopic dermatitis presents a dry and dull state. The skin with xerosis is prone to losing moisture due to weakened skin barrier function, making it easy for external irritant substances to invade the skin.

[0055] At the acute stage of atopic dermatitis, there will be erythema with severe itching, raised papules, blisters, scratch marks and exudate, and secondary infections may occur at this time. As the disease progresses, at the subacute stage, scratches and red papules covered with scales may appear. When entering the chronic stage, repeated scratching of the skin will lead to lichenification with rough and thickened skin and deepened skin lines, and red or brown nodules (prurigo nodularis, nodular prurigo) with severe itching may appear, but not limited to this.

[0056] In this specification, the term "urticaria" is a disease manifested by itchy rash and / or angioedema symptoms, and "chronic urticaria" is defined as urticaria that persists or recurs for more than 6 weeks.

[0057] In this specification, the term "asthma" is a disease in which the bronchi in the lungs are in a highly sensitive state, often causing the bronchi to narrow and resulting in dyspnea, wheezing sounds, and severe coughing. It is an allergic disease triggered by bronchial allergic inflammatory reactions, including bronchial asthma, allergic and non-allergic asthma.

[0058] Allergic rhinitis and bronchial asthma are typical respiratory allergic diseases. Allergic rhinitis is an upper respiratory inflammatory disease, and asthma is a lower respiratory inflammatory disease. Both of these diseases are IgE-mediated inflammatory diseases with airway hyperreactivity and enhanced reactivity to stimulants. At the same time, they have common inflammatory cells such as eosinophils, mast cells, and Th2 cells, and also produce common inflammatory factors such as histamine, leukotrienes, and Th2 cytokines. Therefore, it shows that these two diseases are closely related.

[0059] Allergic rhinitis can be one of the most common chronic inflammatory diseases in adults and children. The nasal mucosa shows hypersensitivity to various allergens, and the main symptoms are clear nasal discharge, nasal congestion, nasal itching, sneezing, etc., and can also be accompanied by eye symptoms such as eye itching and congestion. The typical causes of allergic rhinitis are pollen, dust mites, pet hair, etc., and air pollution and sudden temperature changes can exacerbate the symptoms. Known aggravating factors are exercise, allergens or irritants, weather changes, and respiratory viral infections such as colds. After a number of epidemiological and pathophysiological research results confirmed the close association between the two diseases, the concept of "one airway disease" or "united disease" was proposed, believing that these two allergic inflammatory diseases of the upper and lower respiratory tracts are a single airway disease rather than independent diseases. Therefore, it is necessary to check whether rhinitis patients also have asthma, and it is also necessary to simultaneously evaluate whether asthma patients have rhinitis as a concomitant disease, and a comprehensive diagnosis and treatment method is recommended.

[0060] In this specification, the term "dust mite" may refer to a situation where corpses or excretions stimulate the skin and mucous membranes, leading to the secretion of the inflammatory inducer histamine in the body, thereby exacerbating atopic dermatitis.

[0061] In addition to having control, prevention, improvement, and treatment effects on the allergic diseases, the composition of the present invention also exhibits excellent effects on the control, prevention, improvement, and treatment of allergic diseases and their complications by inducing the normalization of gut microbiota altered due to allergic diseases.

[0062] In this specification, the term "treat" refers to all actions of improving or beneficially altering the symptoms of allergic diseases by administering the pharmaceutical composition of the present invention.

[0063] In this specification, the term "prevent" refers to all actions of suppressing the symptoms of allergic diseases or delaying their progression by administering the pharmaceutical composition of the present invention.

[0064] In this specification, the term "comprising as an active ingredient" means adding the strain of this specification, the lysate of the strain, the culture broth, or an extract of the culture broth thereof in an amount capable of exhibiting the said effects, and includes adding various components as auxiliary components for drug delivery and stabilization, etc., to prepare various forms of formulations.

[0065] In a specific example, the composition may contain Lactobacillus plantarum strains accounting for 0.001% to 80% by weight of the total weight of the composition. In addition, the dosage of Lactobacillus plantarum strains can be 0.01 mg to 10,000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 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, the strain can be at 10 3 to 10 16 CFU / g, 10 3 to 10 15 CFU / g, 10 3 to 10 14 CFU / g, 10 3 to 10 13 CFU / g, 10 3 to 10 12 CFU / g, 10 4 to 10 16 CFU / g, 10 4 to 10 15 CFU / g, 10 4 to 10 14 CFU / g, 104 to 10 13 CFU / g, 10 4 to 10 12 CFU / g, 10 5 to 10 16 CFU / g, 10 5 to 10 15 CFU / g, 10 5 to 10 14 CFU / g, 10 5 to 10 13 CFU / g, 10 5 to 10 12 CFU / g, 10 6 to 10 13 CFU / g, 10 6 to 10 12 CFU / g, 10 7 to 10 13 CFU / g, 10 7 to 10 12 CFU / g, 10 8 to 10 13 CFU / g or 10 8 to 10 12 The content of CFU / g is included, or included in the composition in an equivalent amount of live or dead bacterial culture. Specifically, for adult patients, it can be administered once or in multiple batches, 1×10 3 to 1×10 16Live or dead bacteria at CFU / g. However, the dosage can be prescribed in various ways according to factors such as the formulation method, administration method, age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. Those skilled in the art can consider these factors and appropriately adjust the dosage. The number of administrations can be once, or more than twice within the clinically acceptable range of side effects. The administration site can also be one or more than two sites. For animals other than humans, the same dosage per kg (body weight) as that for humans can also be used, or for example, the dosage can be administered in an amount converted using the volume ratio (e.g., average value) of the target animal to human organs (such as the heart). The administration routes that can be used include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intra-arterial, intraperitoneal, intrathecal, or intravenous), rectal, topical (including transdermal), inhalation, and injection, or the insertion of an implantable device or substance. According to a specific example, the target animals to be treated can be exemplified by humans and mammals for other purposes, specifically including humans, monkeys, mice, rats, rabbits, sheep, cows, dogs, horses, pigs, etc. According to an embodiment, the composition contains inactivated dried strains, and 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 can be administered at one time, and it can be administered 1 to 3 times a day.

[0066] In yet another aspect, there is provided a food composition comprising a Lactobacillus plantarum strain, a lysate, a culture broth derived from the strain, or a mixture thereof as an active ingredient.

[0068] In yet another aspect, there is provided a health functional food for preventing or improving allergic diseases, the health functional food comprising a Lactobacillus plantarum strain, a lysate, a culture broth derived from the strain, or a mixture thereof as an active ingredient.

[0069] In a specific example, the food composition or the health functional food may have anti-allergic activity.

[0070] The "strain", "anti-allergic activity", and "allergic disease" are as described above.

[0071] In a specific example, the improvement of the allergic disease includes the remission of immune allergic reactions or the improvement of skin conditions caused by immune allergic reactions.

[0072] In a specific example, the improvement of the allergic disease may include inhibiting the production of Th2-mediated cytokines, inhibiting the degranulation activity of mast cells, or reducing the levels of serum IgE or MCPT-1.

[0073] In a specific example, the allergic disease may include an allergic reaction caused by house dust, fungi, mites, or the hair, skin, or excrement of animals.

[0074] In a specific example, the allergic disease may be selected from edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic otitis media, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, rash, dry eye, pruritus, anaphylactic shock, pollen allergy, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0075] In a specific example, the food composition may be used for preventing or improving allergic diseases.

[0076] In the food composition, the foods include meat, sausage, bread, chocolate, confectionery, snacks, biscuits, cereals, baked goods, dairy products, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various milk powders, infant formula, milk, milk powder, yogurt, cheese, fermented milk, soup, beverages, tea, instant drinks, alcoholic beverages, vitamin complexes, food additives, functional health foods, and health foods, etc., including all foods in the conventional sense.

[0077] The so-called health functional food (functional food) is the same term as food for special health use (FoSHU), which refers to a food with high medical and therapeutic effects that can effectively exert biological regulatory functions in addition to providing nutrition through processing. Among them, "functional" means obtaining beneficial effects for health purposes on the structure and function of the human body, such as regulating nutrients or physiological functions. The food of the present application can be prepared by methods commonly used in the art, and raw materials and components commonly added in the art can be added during the preparation. In addition, as long as the dosage form of the food is recognized as a food dosage form, it can be prepared without limitation. The food composition of the present application can be made into various types of dosage forms. Different from ordinary drugs, it uses food as the raw material, has the advantages of no side effects when taken for a long time, and excellent portability. Therefore, the HD-02 of the present application can be taken as a supplement to enhance the effect of preventing or improving allergies.

[0078] The so-called health food refers to a food that has a more positive effect on maintaining or promoting health compared to ordinary food, and a health supplement food refers to a food aimed at assisting health. In some cases, the terms health functional food, health food, and health supplement food can be used interchangeably.

[0079] Specifically, the health functional food refers to a food obtained by adding a composition according to an example to food raw materials such as beverages, teas, spices, chewing gums, biscuits, etc., or made into capsules, powders, suspensions, etc. When ingested, it brings specific health effects. Different from ordinary drugs, it uses food as the raw material and has the advantages of no side effects when taken for a long time.

[0080] The food composition according to an example can be taken daily, so a high effect can be expected for the prevention or improvement of allergic diseases, and thus it can be used very effectively.

[0081] The food composition may further contain a physiologically acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the art can be used.

[0082] In addition, the food composition may contain additional ingredients commonly used in food compositions to enhance odor, taste, visual appearance, etc. For example, it may contain vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may also contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), chromium (Cr), etc. In addition, it may also contain amino acids such as lysine, tryptophan, cysteine, valine, etc.

[0083] In addition, the food composition may contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), fungicides (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar pigments, etc.), color developers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), flavoring agents (monosodium glutamate, sodium glutamate, etc.), sweeteners (dulcin, sodium cyclamate, saccharin, sodium, etc.), spices (vanillin, lactones, etc.), leavening agents (alum, potassium bitartrate, etc.), fortifiers, emulsifiers, thickeners (pastes), film-forming agents, gum base materials, defoaming agents, solvents, modifiers, etc. The additives can be screened according to the type of food and used in appropriate amounts.

[0084] The composition according to an example can be added directly or used in combination with other foods or food ingredients, and can be appropriately applied according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (prevention, health care or therapeutic treatment). Generally, when manufacturing food or beverages, the food composition of the present application can be added to food or beverages in an amount of 50 parts by weight or less, specifically 20 parts by weight or less. However, if ingested for a long time for health and hygiene purposes, it may contain a content lower than the above range, and since there are no problems in terms of safety, the active ingredient can also be used in an amount higher than the above range.

[0085] An example of the food composition can be used as a health beverage composition. In this case, it can contain various flavors or natural carbohydrates, etc. as additional ingredients like ordinary beverages. The natural carbohydrates can be monosaccharides such as glucose, fructose; disaccharides such as maltose, sucrose; polysaccharides such as dextrin, cyclodextrin; sugar alcohols such as xylitol, sorbitol, erythritol. Sweeteners can use natural sweeteners such as thaumatin, stevia extract; synthetic sweeteners such as saccharin, aspartame. In every 100 mL of the health beverage composition of the present application, the proportion of the natural carbohydrates can generally be about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g.

[0086] In addition to the above, the healthy beverage composition may further contain various nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, pectates, alginic acid, alginates, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerol, alcohol, or carbonating agents, etc. In addition, it may also contain the pulp used for manufacturing natural fruit juice, fruit juice beverage or vegetable beverage. These components can be used alone or in combination. Although the proportion of these additives is not crucial, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the healthy beverage composition of the present application.

[0087] If the food composition according to an example can exhibit the effect of preventing or improving allergic diseases, it may contain various weight percentages. For example, HD-02 according to the example described can contain 0.0001 to 100% by weight, 0.0001 to 80% by weight, 0.0001 to 50% by weight, 0.0001 to 40% by weight, 0.0001 to 20% by weight, 0.00001 to 10% by weight, 0.0001 to 5% by weight, 0.0001 to 2% by weight, 0.0001 to 1% 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, but not limited thereto, and can contain conventional weight percentages in the art without limitation. Specifically, the fermented milk may contain 0.0001% to 0.01% by weight, 0.0001% to 0.005% by weight, 0.0001% to 0.003% by weight, 0.0001% to 0.002% by weight, or 0.0001% to 0.001% by weight of HD-02 relative to the total weight of the food composition. Specifically, the food beverage may contain 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 of HD-02 relative to the total weight of the food composition. Specifically, the powder or solid food may contain 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 of HD-02 relative to the total weight of the food composition. The mixing amount of the active ingredient can be appropriately determined according to its use purpose (prevention, health care or symptom relief). For example, it may contain Lactobacillus plantarum HD-02 at a concentration of 10 3 CFU / g to 10 16 CFU / g, but not limited thereto.

[0088] The health functional food composition can use the strain or its culture solution alone, or in combination with other foods or food ingredients, and can be appropriately applied according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health care or therapeutic treatment). Generally, when manufacturing food or beverage, the composition of this specification can be added to the raw materials in an amount of 15 parts by weight or less. The type of the health functional food is not particularly limited. Among the types of health functional foods, the beverage composition can contain various flavors or natural carbohydrates, etc. as additional ingredients like ordinary beverages. The natural carbohydrates are 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. As the sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The health food composition can also contain nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerol, alcohol, carbonating agents for carbonated beverages, or combinations thereof. The health functional food composition can also contain the pulp for manufacturing natural fruit juices, fruit juice beverages, vegetable beverages, or combinations thereof.

[0089] In yet another aspect, there is provided a pharmaceutical composition for treating or preventing allergic diseases, the composition comprising a Lactobacillus plantarum strain, a lysate, a culture solution or a mixture thereof derived from the strain as an active ingredient.

[0090] The “strain”, “anti-allergic activity” and “allergic disease” are as described above.

[0091] In a specific example, the pharmaceutical composition can have anti-allergic activity.

[0092] In a specific example, the anti-allergic activity can include inhibiting the degranulation of activated mast cells. Specifically, it can inhibit the secretion of β-hexosaminidase, histamine, prostaglandin and serine protease by activated mast cells.

[0093] In a specific example, the anti-allergic activity can include reducing the concentration of serum IgE or decreasing the binding of IgE to the surface receptor of mast cells. In addition, the anti-allergic activity can include inhibiting the level of serum MCPT-1.

[0094] In a specific example, the allergic disease can include allergic reactions caused by house dust, fungi, mites or the hair, skin or excreta of animals.

[0095] In a specific example, the allergic disease can be selected from edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic otitis media, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, rash, dry eye, pruritus, anaphylactic shock, pollen allergy, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

[0096] The pharmaceutical composition further comprises a pharmaceutically acceptable diluent or carrier. The diluent can be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and the lubricant can be magnesium stearate, talc powder, or a combination thereof. The carrier can be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient can be amorphous cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant can be calcium carboxymethylcellulose, sodium starch glycolate, anhydrous calcium dihydrogen phosphate, or a combination thereof. The binder can be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant can be magnesium stearate, silica, talc powder, or a combination thereof.

[0097] The pharmaceutical composition can be formulated into an oral or non-oral dosage form. The oral dosage form can be granules, powders, liquids, tablets, capsules, dry syrups, or a combination thereof. The non-oral dosage form can be an injection.

[0098] In yet another aspect, there is provided a cosmetic composition for preventing or improving allergic diseases, the composition comprising a Lactobacillus plantarum strain, a lysate, a culture broth, or a mixture thereof derived from the strain as an active ingredient.

[0099] The "strain", "anti-allergy activity", and "allergic disease" are as described above.

[0100] In a specific example, the allergic disease can be a skin allergic disease.

[0101] In a specific example, the cosmetic composition can have anti-allergy activity.

[0102] The cosmetic composition may have, for example, a lotion, a nutritious lotion, a massage cream, a nutritious cream, a serum, a facial mask, a gel, an ampoule, or a skin-adhering cosmetic dosage form.

[0103] The ingredients contained in the cosmetic composition are used as active ingredients, and in addition to the composition, ingredients conventionally used in cosmetic compositions may also be included. For example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances may be included.

[0105] In yet another aspect, a feed composition for preventing or improving allergic diseases is provided, which contains Lactobacillus plantarum strains, lysates, culture broths, or mixtures thereof derived from the strains as active ingredients.

[0106] In a specific example, the feed composition may have anti-allergic activity.

[0107] The "strains", "anti-allergic activity", and "allergic diseases" are as described above.

[0108] The "feed" may refer to any natural or artificially formulated food, a meal of food, etc., or the ingredients of the meal that are used for or suitable for animals to eat, ingest, and digest. There is no particular limitation on the type of feed, and feeds commonly used in the technical field can be used. Non-limiting examples of the feed include plant-based feeds such as grains, roots, food processing by-products, algae, fibrous substances, pharmaceutical by-products, oils, starches, meals, or grain by-products, etc.; and animal-based feeds such as proteins, fat-free substances, oils, minerals, oils, single-cell proteins, zooplankton, or foods.

[0109] The feed composition can be prepared by adding the mixed strain composition within an appropriate effective concentration range according to various feed preparation methods known in the art, and can be used as a feed additive composition for the purpose of preventing or improving allergic diseases.

[0110] When the composition is prepared as a feed additive, the composition can be made into a highly concentrated liquid, powder, or granule form of 20% to 90%. The feed additive may further contain any one or more of organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid, or phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphates (polymeric phosphates), or natural antioxidants such as polyphenols, catechins, α-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid. When prepared as a feed, the composition can be formulated into a conventional feed form and can simultaneously contain conventional feed ingredients.

[0111] The feed additive can be administered to animals alone or in combination with other feed additives in an edible carrier. In addition, the feed additive can be used as a top dressing, directly mixed into animal feed, or in an oral dosage form independent of the feed for easy administration to animals. When the feed additive is administered separately from animal feed, it can be combined with pharmaceutically acceptable edible carriers known in the art to form immediate-release or sustained-release preparations. These edible carriers can be solid or liquid, for example, corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When using a solid carrier, the feed additive can be in the form of tablets, capsules, powders, lozenges, or sugar-coated tablets, or a top dressing in a non-dispersed form. When using a liquid carrier, the feed additive can be in the form of soft gelatin capsules, or dosage forms such as syrups, suspensions, emulsions, or solutions. In addition, the feed and feed additive may contain adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, solubilizing agents, etc. The feed additive can be added to animal feed by soaking, spraying, or mixing.

[0112] The feed or feed additive is applicable to the diets of various animals including mammals, poultry, and fish. As the mammals, it can be used for pigs, cows, horses, sheep, rabbits, goats, rodents, and laboratory rodents such as mice, hamsters, guinea pigs, and pets (e.g., dogs, cats), etc. As the poultry, it can also be used for chickens, turkeys, ducks, geese, pheasants, and quails, etc. As the fish, it can be used for carps, crucian carps, and trouts, etc., but not limited thereto.

[0114] In yet another aspect, there is provided a method for preventing, improving, or treating an individual's condition, the method comprising the step of administering an effective amount of the composition to a desired individual.

[0115] In a specific example, the condition of the individual can be a condition related to IgE (Immunoglobulin E)-mediated diseases or disorders, or allergic diseases.

[0116] Administration can be carried out by methods known in the art. Administration can be directly administered to the individual by any route, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous injection, etc. The administration can be systemic or local administration.

[0117] The individual can be a mammal, for example, a human, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat. The individual can be an individual in need of improving an inflammation-related condition or a bacteria-infection-related condition.

[0118] The administration can be to administer a composition according to a specific example to an individual at 0.00001 mg to 1,000 mg 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, the dose can be variously prescribed according to factors such as the formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, etc. Those skilled in the art can appropriately adjust the dose considering these factors. The number of administrations can be once a day, or can be more than twice within the range of clinically acceptable side effects. For the administration site, it can also be administered at one or more than two sites, once a day or every 2 to 5 days. The total number of days of administration in a single treatment can be 1 to 30 days. When necessary, the same treatment can be repeated after an appropriate period. For animals other than humans, the same dose per kg as for humans can also be administered, or the dose converted, for example, using the volume ratio (such as the average value) of the target animal to human organs (such as the heart, etc.). Effect of the Invention

[0119] A strain according to a protocol and its derived lysate, culture solution, extract of the culture solution, or a mixture thereof has an effect that can be effectively used for preventing, improving, or treating allergic diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0120] Figure 1 It is a figure showing the effect of inhibiting the degranulation of RBL-2H3 mast cells by the supernatant of L. Plantarum HD-02 culture.

[0121] Figure 2 It is a figure showing the effect of inhibiting allergic reactions by L. plantarum HD-02 strain in a mouse model of passive cutaneous allergic reaction induced by anti-DNP IgE.

[0122] Figure 3 It is a figure showing the effect of inhibiting ear swelling by L. plantarum HD-02 strain in a mouse model of atopic dermatitis induced by MC903.

[0123] Figure 4It is a figure showing the effect of L. plantarum HD-02 strain in reducing serum IgE in a mouse model of atopic dermatitis induced by MC903.

[0124] Figure 5 It is a figure showing the effect of L. plantarum HD-02 strain in reducing serum MCPT-1 in a mouse model of atopic dermatitis induced by MC903.

[0125] Figure 6 It is a figure showing the effect of L. plantarum HD-02 strain in alleviating the symptoms of atopic dermatitis in a mouse model of atopic dermatitis induced by house dust mites.

[0126] Figure 7 It is a figure showing the effect of L. plantarum HD-02 strain in reducing serum IgE in a mouse model of atopic dermatitis induced by house dust mites.

[0127] Figure 8 It is a figure showing the effect of L. plantarum HD-02 strain in reducing transepidermal water loss in a mouse model of atopic dermatitis induced by house dust mites.

[0128] Figure 9 It is a figure showing the effect of L. plantarum HD-02 strain in alleviating itching in a mouse model of atopic dermatitis induced by house dust mites.

[0129] Figure 10 It is a figure showing the effect of L. plantarum HD-02 strain in reducing the number of B cells, T cells and total cells in axillary lymph nodes in a mouse model of atopic dermatitis induced by house dust mites.

[0130] Figure 11 It is a figure showing the effect of L. plantarum HD-02 strain in reducing Th1 and Th2 cells in axillary lymph nodes in a mouse model of atopic dermatitis induced by house dust mites.

[0131] Figure 12 It is a figure showing the effect of L. plantarum HD-02 strain in reducing skin thickness in a mouse model of atopic dermatitis induced by house dust mites.

[0132] Figure 13 It is a figure showing the effect of L. plantarum HD-02 strain in inhibiting mast cell infiltration and degranulation in a mouse model of atopic dermatitis induced by house dust mites. Detailed implementation mode

[0133] The following preferred embodiments are given to facilitate the understanding of the present invention. However, the embodiments provided below are only for the easier understanding of the present invention, rather than limiting the content of the present invention. The embodiments can be variously changed, and the embodiments are not limited to the embodiments disclosed below and can also be implemented in various forms.

[0135] Example 1. Isolation and identification of Lactiplantibacillus plantarum strains

[0136] 1.1. Isolation of strains

[0137] For the isolation of Lactiplantibacillus plantarum strains, various pickles obtained from households were used. 25 g of pickle samples were diluted in 225 mL of 0.85% NaCl solution, placed in a sample bag with a wire mesh filter in a homogenizer, and homogenized with a Stomacher for 5 minutes. After the filtrate was serially diluted tenfold, it was spread on MRS plate medium (pH 5.5, Difco, USA) and anaerobically cultured at 37 °C for 48 hours. After cultivation, single colonies were distinguished by morphology and color and purified by three subcultures. Strains that were positive and negative were screened out by Gram staining and catalase tests, respectively, and arbitrarily named HD-02, and finally the Lactiplantibacillus plantarum HD-02 (hereinafter referred to as "HD-02") strain was selected.

[0139] 1.2. Molecular biological identification of the selected strains

[0140] For the 16S rRNA base sequence analysis of the HD-02 strain, bacterial universal primers were used for 16S rRNA gene amplification, and then the base sequence was analyzed (Macrogen Inc, Korea). PCR was performed using the primer sequences shown in Table 1, and the analysis result was the same as SEQ ID NO: 1.

[0141] Table 1

[0142] The 16S rRNA base sequences of each strain were analyzed using the sequence alignment program (BLAST) of the National Center for Biotechnology Information (NCBI) in the United States. The results showed that the HD-02 strain had more than 99% identity with the 16S rRNA genes of multiple strains belonging to the genus Lactiplantibacillus, including the standard strain (JCM1149) of Lactiplantibacillus plantarum (Table 2).

[0143] Table 2

[0144] Since the 16S rRNA base sequence of Lactiplantibacillus plantarum has more than 99% identity with related species such as Lactiplantibacillus pentosus, for more accurate identification, in addition to the 16S rRNA gene, another taxonomic biomarker, the recA gene, was also analyzed. Gene amplification was performed using the recA-specific primers shown in Table 3, and then the base sequence was analyzed (Bionics Co., Ltd, Korea). The analysis results were the same as SEQ ID NO: 2.

[0145] Table 3

[0146] The base sequences of the recA genes of each strain were analyzed using the sequence alignment program (BLAST) of the National Center for Biotechnology Information (NCBI) in the United States. The results showed that the HD-02 strain had more than 99% identity with the recA gene of Lactiplantibacillus plantarum (Table 4).

[0147] Table 4

[0148] 1.3. Analysis of the morphological and physiological characteristics of the selected strains

[0149] The morphological and physiological characteristics of strain HD-02 were determined using the content of Bergey's manual of systematic bacteriology and the API 50CHL kit (Biomerieux SA, France). The morphological and physiological characteristics of strain HD-02 are summarized in Table 5, and the sugar fermentation characteristics of the strain are summarized in Table 6.

[0150] Table 5

[0151] Table 6

[0152] Positive: (+) / Negative: (-) The results of the sugar fermentation characteristics analysis showed that strain HD-02 had a 66.8% similarity with Lactiplantibacillus plantarum.

[0153] Based on the above 16S rRNA base sequence, recA gene base sequence analysis results, and morphological and physiological characteristics, the isolated strain was named Lactiplantibacillus plantarum HD-02. Strain HD-02 was deposited at the Korean Agricultural Genebank (KACC) on August 26, 2022 (Accession No.: KACC81228BP).

[0154] Experimental Example 1 Inhibitory effect of the culture supernatant of L. plantarum HD-02 on the degranulation of RBL-2H3 mast cells

[0155] The culture supernatants of various L. plantarum strains including strain L. plantarum HD-02 were used to treat activated mast cells that induce allergic reactions, and the inhibitory effect on degranulation was screened by measuring the secretion amount of β-hexosaminidase.

[0156] The RBL-2H3 cells were suspended in MEM medium containing 15% FBS, and then at 1.5×10 per well 5Cells were seeded into 24-well plates and co-cultured with 20 ng / mL DNP-IgE for about 12 hours for sensitization. After washing the cells 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), the culture supernatants of various L. plantarum strains were diluted in PIPES buffer at a ratio of 1:15 and pre-treated in each well for 2 hours. After culturing the L. plantarum strains in MRS medium, the strains were precipitated by centrifugation and only the supernatant was collected, then filtered through a 0.22 μm filter to obtain the culture supernatant. Then, 25 ng / mL DNP-HSA (antigen) was added and stimulated for 15 minutes, and then cooled on ice for 5 minutes to terminate the reaction. The supernatant was transferred to a 1.5 mL microcentrifuge tube (e-tube), washed once with PIPES buffer, and the remaining cells on the plate were lysed with 0.1% triton X-100. 30 μL each of the supernatant and the lysed cell sample were mixed in a 96-well plate and reacted at 37 °C for 1 hour. After adding 0.1 M carbonate buffer to terminate the reaction, the absorbance was measured at a wavelength of 405 nm using a microplate reader, and the results are as Figure 1 shown.

[0157] As Figure 1 shown, the culture supernatants of 14 different L. plantarum strains were screened, and the results showed that the culture supernatant of L. plantarum HD-02 strain had the most significant inhibitory effect on mast cell degranulation.

[0159] Experimental Example 2 Inhibitory effect of the L. plantarum HD-02 strain administration group on IgE-mediated allergic reactions in a passive cutaneous anaphylaxis animal model

[0160] The inhibitory effect of the L. plantarum HD-02 strain on allergic reactions was confirmed in an animal model of passive cutaneous anaphylaxis (PCA) induced by anti-DNP IgE.

[0161] Four-week-old BALB / c mice were purchased. After a 1-week adaptation period, the freeze-dried L. plantarum HD-02 strain was suspended in D-PBS. Starting two days before IgE sensitization and continuing until the end of the experiment, 1×10 10The dose of CFU was administered orally. The positive control group used dexamethasone (Dexa) at a concentration of 10 mg / kg. To induce passive cutaneous anaphylaxis, 20 ng of anti-DNP IgE was locally injected into one ear of the mice. Approximately 24 hours later, DNP-HSA was diluted to a concentration of 0.5 mg / mL in 5 mg / mL Evans blue staining solution, and 200 μL was injected via the tail vein. After 30 minutes, the mice were euthanized, the ears were cut off and placed in 700 μL of formamide, and incubated at 63 °C for 12 hours to extract the dye exuded from the ear tissues. After removing the supernatant, the absorbance was measured at a wavelength of 620 nm. The quantification of the exuded Evans blue was determined using a standard calibration curve, as Figure 2 shown.

[0162] As Figure 2 shown, in the passive cutaneous anaphylaxis model induced by anti-DNP IgE, compared with the negative control group (Ag), the experimental group administered with L. plantarum HD-02 strain (Ag+HD-02) inhibited the allergic reaction by approximately 20.4%, thus confirming its anti-allergic effect.

[0164] Experimental Example 3 Inhibitory effect of the L. plantarum HD-02 strain administration group on allergic reactions in an atopic dermatitis animal model induced by MC903

[0165] 3.1. Ear swelling inhibitory effect of the L. plantarum HD-02 strain administration group

[0166] The ear swelling inhibitory effect of the L. plantarum HD-02 strain was confirmed in an atopic dermatitis animal model induced by MC903.

[0167] Six-week-old BALB / c mice were purchased. After a 1-week adaptation period, the freeze-dried L. plantarum HD-02 strain was suspended in D-PBS, and from 7 days before inducing atopic dermatitis until the end of the experiment, each mouse was administered orally at a dose of 2×10 9 CFU per day. To induce atopic dermatitis, after fixing the mice, in the disease induction group, 20 μL of 2 nmol MC903 (calcipotriol, C27H40O3) dissolved in ethanol was applied to both ears of each mouse every day for 12 consecutive days, and the normal group was only applied with ethanol. To confirm the degree of ear swelling induced by atopic dermatitis, the thickness of the mice's ears was measured at the same time every day, and the results were as Figure 3 shown. At this time, the change in ear swelling was calculated by subtracting the ear thickness measured before disease induction from the ear thickness measured every day after disease induction.

[0168] As Figure 3As shown, in the atopic dermatitis model induced by MC903, compared with the control group (MC903), an effect of suppressing ear swelling by approximately 48.4% was confirmed in the experimental group (MC903 + HD-02) administered with the L. plantarum HD-02 strain.

[0170] 3.2. Effect of reducing total serum IgE in the group administered with the L. plantarum HD-02 strain

[0171] Overexpression of IgE is the most typical hallmark of atopic dermatitis. To confirm the total serum IgE concentration in the MC903-induced atopic dermatitis animal model of Experimental Example 3.1, it was measured using a mouse IgE ELISA kit (Biolegend #432404).

[0172] At 4°C, the capturing antibody diluted in coating buffer was reacted overnight in microwell plates, then washed 4 times with washing buffer, and the assay diluent was used to block for 1 hour at room temperature. After washing 4 times, standard samples and serum samples diluted 1:200 were added, reacted for 2 hours at room temperature, then washed 4 times, the detection antibody was added, and reacted for 1 hour at room temperature. After washing 4 times, avidin-HRP was added, reacted for 30 minutes at room temperature, then washed 5 times, the TMB substrate solution was added, and reacted for 20 minutes at room temperature in the dark. The reaction termination solution was added to each well to terminate the color reaction, and the absorbance was measured at a wavelength of 450 nm using an ELISA reader. The results are as Figure 4 shown.

[0173] As Figure 4 shown, compared with the normal group, the total serum IgE concentration increased sharply in the control group (MC903), while an effect of reducing the total serum IgE concentration by approximately 57.2% was confirmed in the experimental group (MC903 + HD-02) of the L. plantarum HD-02 strain.

[0175] 3.3. Effect of reducing serum MCPT-1 in the group administered with the L. plantarum HD-02 strain

[0176] To confirm the concentration of serum mast cell protease-1 (MCPT-1) in the MC903-induced atopic dermatitis animal model of Experimental Example 3.1, a mouse MCPT-1 (mMCP-1) ELISA kit (Invitrogen #88-7503) was used for determination.

[0177] At 4°C, the capture antibody diluted in coating buffer was incubated overnight in the microplate wells, then washed 3 times with washing buffer, and blocked with ELISA / ELISPOT diluent at room temperature for 1 hour. After washing 3 times, standard samples and 1:100 diluted serum samples were added, and the reaction was carried out at room temperature for 2 hours. Then, after washing 3 times, the detection antibody was added, and the reaction was carried out at room temperature for 1 hour. After washing 3 times, avidin-HRP was added, and the reaction was carried out at room temperature for 30 minutes. Then, after washing 5 times, the TMB substrate solution was added, and the reaction was carried out in the dark at room temperature for 15 minutes. The reaction stop solution was added to each well to terminate the color reaction, and the absorbance was measured at a wavelength of 450 nm using an ELISA reader. The results are as Figure 5 shown.

[0178] As Figure 5 shown, compared with the normal group, the serum MCPT-1 concentration increased sharply in the control group (MC903), while in the experimental group of L. plantarum HD-02 strain (MC903 + HD-02), an effect of reducing the serum MCPT-1 concentration by about 56.8% was confirmed.

[0180] Experimental Example 4 Inhibitory effect of the L. plantarum HD-02 strain administration group on allergic reactions in a house dust mite-induced atopic dermatitis animal model

[0181] 4.1. Efficacy of relieving atopic dermatitis symptoms in the L. plantarum HD-02 strain administration group

[0182] The inhibitory effect of the L. plantarum HD-02 strain on atopic dermatitis symptoms was confirmed in a house dust mite-induced atopic dermatitis animal model.

[0183] Six-week-old NC / Nga mice were purchased. After a one-week adaptation period, the hair on the mice's backs up to the upper part of the auricles was shaved off completely. To remove the skin fat component and disrupt the stratum corneum barrier, 150 μL of 4% SDS aqueous solution was sprayed on the depilated area. After it was completely dry, 100 mg of Biostar AD (a natural mite-derived component, Dermatophagoides farinae) ointment was evenly applied. The inducer was applied twice a week for 3 weeks to induce atopic dermatitis, and then once a week for 8 weeks to maintain atopic dermatitis. Administration of L. plantarum HD-02 strain started after the end of the 3-week atopic dermatitis induction period and continued until the end of the experiment. It was orally administered at a dose of 2×10 9 CFU per mouse per day. The positive control group, dexamethasone, was applied to the back skin at a concentration of 0.1% at a dose of 100 μL per mouse twice a week.

[0184] Severe atopic dermatitis symptoms were evaluated using the sum of scores (DermatitisScore) of the following 4 indicators once a week. The evaluation items were erythema / hemorrhage, scarring / dryness, edema, and excoriation / erosion. Each item was scored from 0 (asymptomatic), 1 (mild), 2 (moderate), to 3 (severe), and then totaled. The results were as Figure 6 shown.

[0185] As Figure 6 shown, the severe atopic dermatitis score of the control group (dust mite-induced) was measured to be an average of 8.17 points, and the score of the positive control group (Dexa) was measured to be 2.50 points. Compared with the control group, it showed an atopic dermatitis symptom remission effect of approximately 69.4%. In addition, the severe atopic dermatitis score of the experimental group (dust mite-induced + HD-02) was measured to be an average of 3.00 points, showing an atopic dermatitis symptom remission effect of approximately 63.3%, and it was confirmed to be similar to the level of the positive control group (Dexa).

[0187] 4.2. Effect of L. plantarum HD-02 strain administration group on reducing total serum IgE

[0188] To confirm the total serum IgE concentration in the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1, it was measured using a mouse IgE ELISA kit by the same method as in Experimental Example 3.2. The results were as Figure 7 shown.

[0189] AsFigure 7 As shown, compared with the normal group (no mite induction), the serum total IgE concentration in the control group (mite induction) increased sharply. Compared with the control group (mite induction), the positive control group (Dexa) showed an inhibitory effect on serum total IgE of approximately 63.7%, and the experimental group (mite induction + HD-02) showed an inhibitory effect of approximately 50.6%.

[0191] 4.3. Effect of L. plantarum HD-02 strain administration group on transepidermal water loss

[0192] To confirm the transepidermal water loss caused by impaired skin barrier function in the mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1, the Tewameter TM Nano device was used to measure the water evaporation rate on the skin surface, and the results are as Figure 8 shown.

[0193] As Figure 8 shown, compared with the normal group (no mite induction), transepidermal water loss in the control group (mite induction) increased sharply. Compared with the control group, the positive control group (Dexa) showed a reduction in transepidermal water loss value of approximately 51.3%, and the experimental group (mite induction + HD-02) showed a reduction of approximately 36.9%. Thus, the skin barrier improvement effect of L. plantarum HD-02 was confirmed.

[0195] 4.4. Itching relief effect of L. plantarum HD-02 strain administration group

[0196] From the mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1, it was confirmed that the itching sensation, an important evaluation index for verifying the disease model - the itching degree of atopic skin lesion sites, could be verified. Each group of mice was videotaped for 10 minutes, and then the scratching behavior frequency of the mice in the video was measured. The results are as Figure 9 shown.

[0197] As Figure 9 shown, compared with the normal group (no mite induction), the number of scratching times in the control group (mite induction) increased by approximately 18.7 times. Compared with the control group, an itching relief effect of approximately 51.5% was confirmed in the positive control group (Dexa) and approximately 55.2% in the experimental group (mite induction + HD-02).

[0199] 4.5. Confirm the change in cell composition in the axillary lymph node of the L. plantarum HD-02 strain administration group

[0200] Changes in the composition of immune cells in the important immune tissue - axillary lymph node of the disease model were confirmed in the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1.

[0201] Axillary lymph nodes of each mouse were extracted to obtain single-cell suspensions, and then the cell numbers were determined and aliquoted at 1.0 x 10 6 cells per well into a 96-well U-bottom microwell plate. Treated with eBioscience TM Cell Stimulation Cocktail (plus protein transport inhibitors) and after 4 hours of stimulation, stained with anti-CD3-Pacific blue, anti-CD4-PE-Cy7, anti-IL-10-FITC, anti-IL-4-PE, anti-IFN-γ-APC antibodies, and subjected to FACS analysis. The results are as Figure 10 and Figure 11 shown.

[0202] As Figure 10 shown, compared with the normal group (without dust mite induction), the total cell number in the axillary lymph nodes of the control group (dust mite-induced) with induced atopic dermatitis increased by about 7.5 times. Compared with the control group, it was confirmed that the number of axillary lymph node cells decreased by about 53.7% in the positive control group (Dexa) by about 81.7% and in the experimental group (dust mite-induced + HD-02). In addition, as Figure 11 shown, compared with the control group, after administration of L. plantarum HD-02, it was confirmed that B cells, T cells, Th1, and Th2 cells in the axillary lymph nodes also decreased significantly.

[0204] 4.6. Confirm the skin thickness and histological changes in the L. plantarum HD-02 strain administration group

[0205] The effects of L. plantarum HD-02 strain on the increased skin thickness and mast cell infiltration caused by the inflammatory response were confirmed in the dust mite-induced atopic dermatitis model (severe atopic dermatitis model) of Experimental Example 4.1.

[0206] The back skin of the mice was excised, and the tissues were fixed in 10% aqueous formaldehyde solution. Then, paraffin blocks were prepared and sliced into sections with a thickness of 4 μm to make slides. The accumulation of lymphocytes in the skin tissues and the changes in the epidermal thickness were observed by hematoxylin & eosin (H&E) staining, and the number of mast cells was measured by toluidine blue staining to confirm the degree of tissue infiltration. The results are shown as Figure 12 and Figure 13 follows.

[0207] As Figure 12 shown, compared with the normal group (without dust mite induction), the thickness of the epidermal layer in the control group (induced with dust mites) with atopic dermatitis was increased by about 8.8 times. Compared with the control group, the epidermal layer thickness was inhibited by about 56.7% in the positive control group (Dexa) and about 57.3% in the experimental group (induced with dust mites + HD-02). In addition, compared with the control group, the degree of lymphocyte accumulation in the inflammatory tissues of the experimental group was significantly reduced to the level of the positive control group (Dexa).

[0208] In addition, as Figure 13 shown, the results of observing the infiltration of mast cells in the inflammatory tissues confirmed that compared with the normal group, the number of mast cells in the control group with atopic dermatitis was increased by about 6.2 times. Compared with the control group, the number of mast cells was significantly reduced by about 61.4% in the positive control group (Dexa) and about 47.0% in the experimental group.

[0210] The above results indicate that the L. plantarum HD-02 strain can be effectively applied for the prevention, improvement, or treatment of atopic dermatitis or allergic diseases.

[0212] The above description of the present invention is for illustration purposes. Those skilled in the art should understand that without changing the technical idea or essential features of the present invention, the present invention can be easily modified into other specific forms. Therefore, it should be understood that the above-described embodiments are exemplary in all aspects and not restrictive.

[0213] [Deposit Number]

[0214] Name of the depositary institution: Agricultural Genebank (abroad)

[0215] Deposit number: KACC81228BP

[0216] Date of deposit: 20220826

Claims

1. An anti-allergy composition, wherein, The composition contains Lactiplantibacillus plantarum strain HD-02 deposited under the accession number KACC81228BP, a lysate, a culture broth, or a mixture thereof derived from the strain as an active ingredient.

2. The anti-allergy composition according to claim 1, wherein The composition inhibits the production of Th2 (Helper TCellType 2)-mediated cytokines.

3. The anti-allergy composition according to claim 1, wherein The composition reduces the concentration of serum IgE or decreases the binding of IgE to the surface receptor of mast cells.

4. The anti-allergy composition according to claim 1, wherein, The composition further contains one or more additives selected from preservatives, dyes, emulsifiers, sweeteners, stabilizers, flavor enhancers, fragrances, and acidulants.

5. The anti-allergy composition according to claim 1, wherein, The composition is selected from dosage forms such as solutions, emulsions, viscous mixtures, powders, granules, tablets, and capsules.

6. A food composition, wherein, The composition contains Lactiplantibacillus plantarum strain HD-02 deposited under the accession number KACC81228BP, a lysate, a culture broth, or a mixture thereof derived from the strain as an active ingredient.

7. The food composition according to claim 6, wherein, The food is selected from ice cream, infant formula, milk, yogurt, cheese, fermented milk, milk powder, grains, baked goods, dairy products, beverages, and food additives.

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

9. The pharmaceutical composition according to claim 8, wherein, The composition has the activity of inhibiting mast cell degranulation.

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

11. The pharmaceutical composition according to claim 8, wherein, The allergic disease includes allergic reactions caused by house dust, fungi, mites, or the hair, skin, or excrement of animals.

12. The pharmaceutical composition according to claim 8, wherein, The allergic disease is selected from edema, anaphylaxis, rhinitis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, allergic tympanitis, atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, rash, dry eye, pruritus, anaphylactic shock, pollen allergy, 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, wherein, The health-care functional food contains, as an active ingredient, the Lactiplantibacillus plantarum HD-02 strain deposited under the accession number KACC81228BP, a lysate, a culture solution, or a mixture thereof derived from the strain.

14. The healthcare functional food according to claim 13, wherein, The improvement of the allergic disease includes: - inhibiting the production of Th2-mediated cytokines; - inhibiting the degranulation activity of mast cells; - inhibiting the expression of serum IgE; or - inhibiting the level of serum MCPT-1.

15. The health care functional food according to claim 13, wherein, The improvement of the allergic disease includes the alleviation of immune allergic reactions or the improvement of skin conditions caused by immune allergic reactions.

16. The health care functional food according to claim 13, wherein, The allergic disease includes allergic reactions caused by house dust, fungi, mites, or the hair, skin, or excrement of animals.

17. The health care functional food according to claim 13, wherein, The allergic disease is 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, rash, dry eye, pruritus, anaphylactic shock, pollen allergy, bacterial allergy, fungal allergy, viral allergy, insect allergy, food allergy, drug allergy, and respiratory allergy.

18. A cosmetic composition for improving or preventing skin allergic diseases, wherein, The composition contains the Lactiplantibacillus plantarum HD-02 strain deposited under the accession number KACC81228BP, a lysate, a culture solution, or a mixture thereof derived from the strain.

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

20. A method for preventing or treating allergic diseases, wherein, The method includes the step of administering to a desired individual a composition containing an effective amount of the Lactiplantibacillus plantarum HD-02 strain deposited under the accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof.

21. Use of a composition in the preparation of a pharmaceutical preparation for preventing or treating allergic diseases, wherein, The composition contains the Lactiplantibacillus plantarum HD-02 strain deposited under the accession number KACC81228BP, a culture of the strain, a lysate of the strain, or a mixture thereof.

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