Composition for promoting interleukin-23 production

Abstract

To provide a composition for promoting interleukin-23 production, capable of saving complicated labor such as taste change and encapsulation.SOLUTION: The composition for promoting interleukin-23 production contains Lactobacillusjohnsonii as an active ingredient. Here, the Lactobacillus johnsonii is preferably Lactobacillus johnsonii OLL203565 (accession number: NITEBP - 02003) or Lactobacillus johnsonii OLL204255 (accession number: NITEBP - 02004).SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition for promoting interleukin-23 production. [Background technology]

[0002] In the past, "interleukin (hereinafter abbreviated as "IL")-23 secretion (production) promoters containing low molecular weight monocarboxylic acids such as acetic acid, lactic acid, or thioglycolic acid as active ingredients" have been proposed (see, for example, JP 2008-127277 A). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Application Laid-Open No. 2008-127277 [Patent Document 2] International Publication No. 2015 / 087919 [Patent Document 3] International Publication No. 2011 / 145737 [Patent Document 4] International Publication No. 2012 / 033151 [Patent Document 5] Special table number 2012-526751 [Patent Document 6] International Publication No. 2016 / 136942 [Non-patent literature]

[0004] [Non-Patent Document 1] Mariman, R et al., British Journal of Nutrition, (2014), Vol. 112, pp 1088-1097 [Non-patent document 2] Cinque, B et al, PLoS ONE, (2016) , 11(9), e0163216 Summary of the Invention [Problem to be solved by the invention]

[0005] However, low-molecular-weight monocarboxylic acids such as acetic acid, lactic acid, and thioglycolic acid have a relatively strong sour taste and are difficult to ingest as they are. Therefore, in order to ingest low-molecular-weight monocarboxylic acids without discomfort, processing such as formulation is required. Specifically, complicated efforts such as adding sweeteners to change the taste or encapsulating the product are required.

[0006] The present invention aims to provide an IL-23 production promoter, a composition for promoting IL-23 production, and the like that can eliminate the need for cumbersome procedures such as changing the flavor or encapsulating the product. [Means for solving the problem]

[0007] A composition for promoting IL-23 production according to a first aspect of the present invention contains, as an active ingredient, a bacterial cell, which is at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactobacillus johnsonii.The lactic acid bacteria used were Lactobacillus delbrueckii subsp. bulgaricus strain 2038, Lactobacillus delbrueckii subsp. bulgaricus strain OLL1181 (accession number: FERM BP-11269), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (accession number: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (accession number: FERM BP-10741), Lactobacillus johnsonii strain OLL203565 (accession number: NITE BP-02003), and Lactobacillus johnsonii strain OLL204255 (accession number: NITE BP-02003). bulgaricus strain OLL1181 (Accession Number: FERM BP-11269), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (Accession Number: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession Number: FERM BP-10741), Lactobacillus johnsonii strain OLL203565 (Accession Number: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession Number: FERM BP-10741), Lactobacillus johnsonii strain OLL203565 (Accession Number: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession Number: FERM BP-10741), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession Number: FERM BP-76), Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession Number: NITE ... BP-02003) and Streptococcus thermophilus strain 1131, and even more preferably at least one selected from the group consisting of Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession Number: NITE BP-76) and Streptococcus thermophilus strain 1131. That is, the above-mentioned bacterial cells are used as a composition for promoting IL-23 production or as one of its components. The bacterial cells may be live or killed. When the bacterial cells are killed, the killed cells are preferably heat-killed, i.e., bacterial cells killed by heat. Furthermore, the "composition" referred to here includes pharmaceuticals, supplements, food additives, and other preparations, foods and beverages (excluding animals and plants themselves).) and food and drink compositions (including processed food and drink) that can be ingested by animals (including humans).

[0008] As a result of extensive investigations by the present inventors, it has been revealed that the bacterial cells according to the first aspect of the present invention can promote the production of IL-23. That is, these bacterial cells can effectively promote the production of IL-23.

[0009] IL-23, for example, acts on ILC3 to induce the production of IL-22. When IL-22 is recognized by small intestinal epithelial cells, it induces the expression of antimicrobial peptides such as Reg3 family proteins. These antimicrobial peptides then kill foreign bacteria and other microorganisms, thereby suppressing the spread of microbial infection and excessive inflammatory responses. However, this effect is merely one example, and it is entirely conceivable that IL-23 may induce the production of other substances and exert other effects.

[0010] Furthermore, these bacterial cells are tasteless. Therefore, when preparing a composition for promoting IL-23 production, it is possible to avoid the troublesome steps of changing the flavor, encapsulating, etc. Of course, the flavor of the composition for promoting IL-23 production may be changed or the composition for promoting IL-23 production may be encapsulated from other perspectives.

[0011] When the dendritic cells are tsDCs, among the above-mentioned bacterial strains, bacterial strains that produce interleukin-23 at a concentration of 30 pg / mL or more in tsDCs are preferred, bacterial strains that produce interleukin-23 at a concentration of 50 pg / mL or more are more preferred, bacterial strains that produce interleukin-23 at a concentration of 70 pg / mL or more are even more preferred, and bacterial strains that produce interleukin-23 at a concentration of 90 pg / mL or more are particularly preferred.

[0012] Furthermore, as mentioned above, when the dendritic cells are tsDCs, specific examples of the bacteria that can be used to stimulate tsDCs include Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. bulgaricus that produce interleukin-23 at concentrations of 30 pg / mL or more, Lactobacillus delbrueckii subsp. bulgaricus that produce interleukin-23 at concentrations of 40 pg / mL or more, Lactobacillus delbrueckii subsp. bulgaricus that produce interleukin-23 at concentrations of 90 pg / mL or more, Streptococcus thermophilus that produce interleukin-23 at concentrations of 40 pg / mL or more, Streptococcus thermophilus that produce interleukin-23 at concentrations of 70 pg / mL or more, and Streptococcus thermophilus that produce interleukin-23 at concentrations of 60 pg / mL or more.

[0013] Furthermore, when the dendritic cells are BMDCs, among the above-mentioned bacterial cells, bacterial cells that produce interleukin-23 at a concentration of 300 pg / mL or more in BMDCs are preferred, bacterial cells that produce interleukin-23 at a concentration of 600 pg / mL or more are more preferred, bacterial cells that produce interleukin-23 at a concentration of 900 pg / mL or more are even more preferred, bacterial cells that produce interleukin-23 at a concentration of 1200 pg / mL or more are even more preferred, bacterial cells that produce interleukin-23 at a concentration of 1500 pg / mL or more are even more preferred, and bacterial cells that produce interleukin-23 at a concentration of 2000 pg / mL or more are particularly preferred.

[0014] Furthermore, as mentioned above, when the dendritic cells are BMDCs, specifically, Lactobacillus delbrueckii subsp. bulgaricus that produces interleukin-23 at a concentration of 800 pg / mL or more in BMDCs, Lactobacillus delbrueckii subsp. bulgaricus that produces interleukin-23 at a concentration of 1000 pg / mL or more, and Lactobacillus delbrueckii subsp. bulgaricus that produces interleukin-23 at a concentration of 1300 pg / mL or more are used. bulgaricus, Lactobacillus delbrueckii subsp. bulgaricus, which produces interleukin-23 at concentrations of 1600 pg / mL or more, Streptococcus thermophilus, which produces interleukin-23 at concentrations of 2000 pg / mL or more, Lactobacillus johnsonii, which produces interleukin-23 at concentrations of 300 pg / mL or more, and Lactobacillus johnsonii, which produces interleukin-23 at concentrations of 800 pg / mL or more.

[0015] A method according to a second aspect of the present invention is a method of using the aforementioned bacterial cell as an IL-23 production promoter, i.e., providing the aforementioned bacterial cell for use as an IL-23 production promoter, excluding the act of treating humans.

[0016] A method according to a third aspect of the present invention is a method for promoting IL-23 production in vivo by orally administering the above-mentioned composition for promoting IL-23 production. However, this does not include the use of the composition for promoting human therapy. The administration period is preferably one week or longer, more preferably two weeks or longer, even more preferably three weeks or longer, and particularly preferably four weeks or longer.

[0017] In a method for promoting IL-23 production according to a third aspect of the present invention, the aforementioned bacterial cells are contacted with dendritic cells. That is, IL-23 production is promoted by contacting the bacterial cells with dendritic cells. However, this does not include the use of the bacterial cells to treat humans.

[0018] The use of the aforementioned bacterial cells according to a fourth aspect of the present invention is for the manufacture of a composition for promoting IL-23 production, excluding the use in treating humans. [Brief explanation of the drawings]

[0019] [Figure 1] FIG. 1 is a bar graph showing the amount of IL-23 secreted (pg / mL) when tsDC was stimulated with heat-killed cells of various lactic acid bacteria. [Figure 2] FIG. 1 is a bar graph showing the amount of IL-23 secreted (pg / mL) when BMDCs were stimulated with heat-killed cells of various lactic acid bacteria. [Figure 3] FIG. 1 is a bar graph showing the amount of IL-23 secreted (pg / mL) when BMDCs were stimulated with heat-killed cells of various lactic acid bacteria and bifidobacteria. DETAILED DESCRIPTION OF THE INVENTION

[0020] Hereinafter, the present invention will be described in detail by showing embodiments of the present invention, but the present invention is not limited to the individual embodiments described below.

[0021] A composition for promoting IL-23 production according to an embodiment of the present invention contains bacterial cells as an active ingredient. The bacterial cells may be live or killed. When the bacterial cells are killed, they are preferably heat-killed. The term "heat-killed bacterial cells" refers to the bacterial cells that remain after being killed by heat. The composition for promoting IL-23 production may consist solely of bacterial cells or may contain other components. Examples of bacterial cells include lactic acid bacteria and bifidobacteria. Lactic acid bacteria refer to all bacteria taxonomically recognized as lactic acid bacteria or related bacteria, without limitation in terms of species or strain. Bifidobacteria refer to all bacteria taxonomically recognized as bifidobacteria or related bacteria, without limitation in terms of species or strain. Lactic acid bacteria are sometimes classified as plant-derived or animal-derived depending on their origin, and both plant- and animal-derived lactic acid bacteria can be used in the present invention.

[0022] In an embodiment of the present invention, the lactic acid bacteria is more preferably at least one selected from the group consisting of Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus gasseri, Streptococcus thermophilus, Lactobacillus plantarum, and Lactobacillus johnsonii.

[0023] When the lactic acid bacteria are Lactobacillus delbrueckii subsp. bulgaricus, they are preferably Lactobacillus delbrueckii subsp. bulgaricus strain 2038, Lactobacillus delbrueckii subsp. bulgaricus strain OLL1181 (accession number: FERM BP-11269), Lactobacillus delbrueckii subsp. bulgaricus strain MEP201701, Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (accession number: NITE BP-76), or Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (accession number: FERM BP-10741). When the lactic acid bacteria are Lactobacillus gasseri, they are preferably Lactobacillus gasseri strain OLL2716 (accession number: FERM BP-6999), Lactobacillus gasseri strain OLL2809 (Accession number: NITE BP-72), Lactobacillus gasseri strain OLL2959 (Accession number: NITE BP-224), or Lactobacillus gasseri strain MEP201801 is preferred; in the case of Streptococcus thermophilus, Streptococcus thermophilus strain 1131 and Streptococcus thermophilus strain MEP201702 are preferred; in the case of Lactobacillus plantarum, Lactobacillus plantarum strain OLL2712 (Accession number: FERM BP-11262) or Lactobacillus plantarum strain MEP201802 is preferred; in the case of Lactobacillus johnsonii, Lactobacillus johnsonii strain OLL203565 (Accession number: NITE BP-6999), Lactobacillus gasseri strain OLL2809 (Accession number: NITE BP-72), Lactobacillus gasseri strain OLL2959 (Accession number: NITE BP-224), or Lactobacillus gasseri strain MEP201801 is preferred; in the case of Streptococcus thermophilus, Streptococcus thermophilus strain 1131 and Streptococcus thermophilus strain MEP201702 are preferred; in the case of Lactobacillus plantarum, Lactobacillus plantarum strain OLL2712 (Accession number: FERM BP-11262) or Lactobacillus plantarum strain MEP201802 is preferred; and in the case of Lactobacillus johnsonii, Lactobacillus johnsonii strain OLL203565 (Accession number: NITE BP-699 BP-02003), Lactobacillus johnsonii strain OLL204255 (Accession number: NITE BP-02004), or Lactobacillus johnsonii strain MEP201803 is preferred.

[0024] In addition, in an embodiment of the present invention, the bifidobacterium is preferably Bifidobacterium bifidum.

[0025] The Bifidobacterium bifidum bacteria is preferably the Bifidobacterium bifidum strain OLB6378 (accession number: NITE BP-31) or the Bifidobacterium bifidum strain MEP201804.

[0026] Lactobacillus delbrueckii subsp. bulgaricus strain 2038 can be obtained by isolation from Bulgarian Yogurt LB81 (registered trademark) manufactured by Meiji Co., Ltd. using standard methods. Lactobacillus delbrueckii subsp. bulgaricus strain MEP201701 can be obtained by isolation from fermented milk produced in the Republic of Bulgaria using standard methods. Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 was internationally deposited under the Budapest Treaty with the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under accession number NITE BP-76 on February 10, 2005 (original deposit date) (transferred from original deposit to deposit under the Budapest Treaty on April 1, 2009). Furthermore, the Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 strain was internationally deposited on February 22, 1999 (domestic deposit date) with the National Institute of Advanced Industrial Science and Technology (National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1 Higashi, Tsukuba City, Ibaraki Prefecture, Japan) (later centralized at the National Institute of Technology and Evaluation (National Institute of Technology and Evaluation, Patent Microorganisms Depositary, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under the accession number FERM BP-10741 in accordance with the Budapest Treaty (transferred from the original deposit to a deposit under the Budapest Treaty on November 29, 2006). Furthermore, the Lactobacillus delbrueckii subsp. bulgaricus OLL1181 strain was internationally deposited on July 16, 2010, under the Budapest Treaty, with the Patent Organism Depositary of the National Institute of Advanced Industrial Science and Technology (Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki Prefecture, Japan) (later centralized at the Patent Microorganism Depositary of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba Prefecture, Japan)) under the accession number FERM BP-11269.Furthermore, the Lactobacillus gasseri strain OLL2716 was internationally deposited on May 24, 1999 (original deposit date) with the National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, Ministry of International Trade and Industry (1-1-3 Higashi, Tsukuba, Ibaraki, Japan) (later centralized at the Patent Microorganisms Depositary, National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan) under the accession number FERM BP-6999 in accordance with the Budapest Treaty (transferred from the original deposit to a deposit under the Budapest Treaty on January 14, 2000). Furthermore, the Lactobacillus gasseri strain OLL2809 was internationally deposited on February 1, 2005 (original deposit date) with the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under the accession number NITE BP-72 in accordance with the Budapest Treaty (transferred from the original deposit to a deposit under the Budapest Treaty on January 18, 2006). The Lactobacillus gasseri strain OLL2959 was internationally deposited under the Budapest Treaty with the National Institute of Technology and Evaluation (NIET) Patent Microorganisms Depositary (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under accession number NITE BP-224 on March 31, 2006 (original deposit date) (transferred from original deposit to Budapest Treaty deposit on November 21, 2007). The Lactobacillus gasseri strain MEP201801 can be obtained by isolation from feces of healthy adults using standard methods. The Streptococcus thermophilus strain 1131 can be obtained by isolation from Bulgaria Yogurt LB81 (registered trademark) manufactured by Meiji Co., Ltd. using standard methods. Streptococcus thermophilus MEP201702 can be isolated from fermented milk produced in Bulgaria by conventional methods. Lactobacillus plantarum OLL2712 was deposited on July 2, 2010, under the Budapest Treaty with the Patent Organism Depositary of the National Institute of Advanced Industrial Science and Technology (Central No. 6, 1-1-1 Higashi, Tsukuba, Ibaraki Prefecture, Japan) (later consolidated at the Patent Microorganism Depositary of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba Prefecture, Japan) under the accession number FERM BP-11262.Lactobacillus plantarum MEP201802 strain can be obtained by isolation from raw milk produced in Hokkaido using conventional methods. Lactobacillus johnsonii OLL203565 strain was internationally deposited on February 3, 2015, with the National Institute of Technology and Evaluation (NITE BP-02003, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan) under the Budapest Treaty. Lactobacillus johnsonii OLL204255 strain was internationally deposited on February 3, 2015, with the National Institute of Technology and Evaluation (NITE BP-02004, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu, Chiba, Japan) under the Budapest Treaty. Lactobacillus johnsonii strain MEP201803 can be obtained by isolation from dogwood flower buds grown in the mountains of Kanagawa Prefecture using conventional methods. Bifidobacterium bifidum strain OLB6378 was deposited on October 26, 2004, with the National Institute of Technology and Evaluation (NIET) Patent Microorganisms Deposit Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) under the Budapest Treaty under accession number NITE BP-31. Bifidobacterium bifidum strain MEP201804 can be obtained by isolation from feces of healthy adults using conventional methods.

[0027] The bacterial cells may be frozen or freeze-dried. Furthermore, the bacterial cells may be in the form of only the bacterial cells, or the bacterial cells may contain other components (e.g., a cryoprotectant or a lyoprotectant), or may be dispersed in various media such as starch or water that have been used as ingredients in pharmaceuticals and foods and beverages.

[0028] The composition for promoting IL-23 production according to an embodiment of the present invention exerts its function by being ingested by mammals, including humans. The term "ingestion" as used herein refers to any route of administration, as long as it is administered into the human body, and can be achieved by any known administration method, such as oral administration, tube administration, or enteral administration. Typical examples of such administration include oral and enteral administration via the digestive tract, with oral ingestion being preferred, and ingestion by eating or drinking being more preferred.

[0029] The composition for promoting IL-23 production according to an embodiment of the present invention is sufficient as long as it contains bacterial cells, and the number of bacterial cells is not particularly limited, but preferably contains 100 million or more bacterial cells per unit package, more preferably 500 million or more bacterial cells, and even more preferably 1 billion or more bacterial cells. The greater the number of bacterial cells, the greater the expected effect, but the upper limit is 10 trillion bacterial cells.

[0030] The weight of a unit package of a composition for promoting IL-23 production according to an embodiment of the present invention is not particularly limited, but when 100 million or more bacterial cells are present per gram of the composition for promoting IL-23 production, the weight is preferably in the range of 10 g to 500 g, more preferably 25 g to 250 g, even more preferably 50 g to 200 g, and most preferably 75 g to 150 g. Furthermore, the above-mentioned unit package may refer not only to a unit package per bag, box, or container, but also to a unit package per serving contained therein, or a unit package per day. It may also be a package containing an amount appropriate for ingestion over multiple days, for example, a week's worth, or a package containing multiple individual packages.

[0031] In an embodiment of the present invention, it is desirable to continuously ingest the composition for promoting IL-23 production for at least 3 weeks, preferably at least 5 weeks, and more preferably at least 8 weeks. Because the composition for promoting IL-23 production can be safely ingested, the period of ingestion is not particularly limited and can be continued indefinitely. From the viewpoint of obtaining a sufficiently effective IL-23 production effect, it is preferable to set the ingestion period at approximately 8 weeks.

[0032] The composition for promoting IL-23 production according to an embodiment of the present invention can be used as a pharmaceutical product or a food or beverage. The pharmaceutical product or food or beverage is useful in that it has the effect of promoting IL-23 production, and can be used, for example, as a pharmaceutical product or a food or beverage for preventing or treating microbial infection. When the composition for promoting IL-23 production according to an embodiment of the present invention is used as a pharmaceutical product or a food or beverage, bacterial cells of a single strain may be used, or bacterial cells of two or more strains may be used in combination.

[0033] When the composition for promoting IL-23 production according to the embodiment of the present invention is used as a medicine or a food or beverage, the state of the composition for promoting IL-23 production is not limited, and it may be in any state, such as a paste, spray-dried product, freeze-dried product, vacuum-dried product, drum-dried product, liquid product dispersed in a medium, diluted product diluted with a diluent, or crushed product obtained by crushing a dried product with a mill or the like.

[0034] Furthermore, the composition for promoting IL-23 production according to the embodiment of the present invention can also be formulated as a health food or a food for the sick. The health food system was established in consideration of domestic and international trends and consistency with the existing system of foods for specified health uses, and covers not only ordinary foods but also foods in the form of tablets, capsules, and the like. The system stipulates two types of foods: foods for specified health uses (individually approved) and foods with nutrient functions (standardized). By administering the composition for promoting IL-23 production according to the embodiment of the present invention to animals such as humans as a food for special dietary uses such as a food for specified health uses or a food with nutrient functions, it becomes possible to prevent, for example, various infections.

[0035] It is preferable that the compositions for promoting IL-23 production according to the embodiments of the present invention be labeled with information about their intended use, efficacy, function, type of active ingredient, type of functional ingredient, and method of administration. The term "labeling" as used herein should be appropriate for each of the following: drugs, quasi-drugs, health foods, foods for specified health uses, nutrient-functional foods, general foods, dietary supplements, health foods, supplements, enteral nutrients, oral cosmetics, and feed. Furthermore, the term "labeling" as used herein includes all labeling intended to inform consumers of the above information. This labeling may be any labeling that evokes or infers the above content, regardless of the purpose, content, object, or medium of display. Examples of such labeling include displaying the above information on product packaging or containers, displaying or distributing the above information in product advertisements, price lists, or transaction documents, or providing information containing these information via electromagnetic means (e.g., the Internet).

[0036] When the product containing the composition for promoting IL-23 production according to the embodiment of the present invention is, for example, a food or beverage, it is preferable that the food or beverage be labeled with, for example, "for promoting IL-23 production," "contains lactic acid bacteria that promote IL-23 production," "for enhancing antimicrobial peptides," "for preventing infections," or other similar labels.

[0037] The wording used for the above-mentioned indication is not limited to the above examples, and may be any wording that has the same meaning as the above. For example, various wordings that inform consumers of the promotion of IL-23 production, the enhancement of antimicrobial peptides, the effect of preventing infections, etc. are acceptable.

[0038] When the composition for promoting IL-23 production according to an embodiment of the present invention is used as a food or beverage, the type of food or beverage is not particularly limited. Examples of such food or beverage products include milk, processed milk, soft drinks, fermented milk, yogurt, cheese, other dairy products, bread, biscuits, crackers, pizza crust, infant formula, liquid food, medical foods, foods such as powdered milk for infants, foods such as powdered milk for pregnant and nursing women, and nutritional foods. In producing such food or beverage products, conventional methods for producing food compositions can be used, such as using the bacterial cells, which are the active ingredient of the composition for promoting IL-23 production according to an embodiment of the present invention, as is or mixing them with other foods or beverages or food ingredients. Furthermore, the form of the food or beverage product is not particularly limited, and any commonly used food or beverage form can be used. For example, the form may be any of solids (including powders and granules), pastes, liquids, suspensions, and the like, but is not limited to these. Milk beverages, fermented milk, soft drinks, jelly drinks, tablets, and powdered foods are more preferred, with yogurt being even more preferred.

[0039] The bacterial cells, which are the active ingredient of the composition for promoting IL-23 production according to the embodiment of the present invention, can be supplemented with water, proteins, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, functional ingredients, food additives, and other ingredients commonly found in foods. In the production of the above-mentioned foods and beverages, protein sources that can be used include proteins or protein-containing raw materials commonly used in food production, such as whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, α-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, egg protein, meat protein, and other animal and plant proteins, as well as hydrolysates thereof. Examples of sugar sources include modified starch (textlin, soluble starch, British starch, oxidized starch, starch esters, starch ethers, etc.) and dietary fiber. Lipid sources include animal fats and oils such as lard, fish oil, and their fractionated, hydrogenated, and interesterified oils; and vegetable fats and oils such as palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, and their fractionated, hydrogenated, and interesterified oils. Vitamins include vitamin A, carotenes, B vitamins, vitamin C, vitamin D, vitamin E, vitamin K, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid. Minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Organic acids include malic acid, citric acid, lactic acid, and tartaric acid. Functional ingredients include oligosaccharides, glucosamine, collagen, ceramide, royal jelly, and polyphenols. Examples of food additives include emulsifiers, stabilizers, thickeners, gelling agents, sweeteners, acidulants, preservatives, antioxidants, pH adjusters, colorants, etc. Various milk-derived components such as butter, milk minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, and lactose are examples of components that can be suitably used in the production of foods and beverages according to embodiments of the present invention.

[0040] These ingredients can be used in combination of two or more. The raw materials may be natural products, processed natural products, synthetic products, and / or foods containing large amounts of these.

[0041] The bacterial cells, which are the active ingredient of the composition for promoting IL-23 production according to an embodiment of the present invention, can be used as a pharmaceutical product for preventing or treating infection. When producing such pharmaceutical products, the bacterial cells can be used as a processed product, either crushed or uncrushed. Furthermore, the bacterial cells used may be of one type or multiple types.

[0042] The amount of bacterial cells in the pharmaceutical product can be determined arbitrarily depending on the purpose and use (prophylactic agent, therapeutic agent, etc.) The content can be, for example, 0.001 to 100% (w / w), particularly 0.1 to 100%, of the total amount, but the present invention is not limited thereto.

[0043] The dosage of pharmaceuticals containing the above-mentioned bacterial cells as an active ingredient can be appropriately determined taking into consideration various factors such as the route of administration, the age, weight, and symptoms of the target animal, including humans. An example of an appropriate dosage is 1 to 1,000 mg / kg / day of the active ingredient, but the present invention is not limited to this. For example, when taking the active ingredient for prophylactic purposes over a long period of time, a dosage lower than the above range may be used. Furthermore, since there are no apparent safety issues with this active ingredient, it is acceptable to use a dosage higher than the above range.

[0044] The dosage form of the pharmaceutical is preferably a dosage form that can be administered orally, so that the bacterial cells of the present invention can reach the intestines. Examples of preferred dosage forms of the pharmaceutical according to the present invention include tablets, coated tablets, capsules, granules, powders, liquids, syrups, and lozenges. These various preparations can be formulated by mixing the bacterial cells as the main drug with auxiliary agents commonly used in the pharmaceutical formulation technology, such as excipients, binders, disintegrants, lubricants, colorants, flavorings, solubilizers, suspending agents, and coating agents, according to conventional methods.

[0045] When the above-mentioned bacterial cells are used as a pharmaceutical, for example, in the case of oral administration, the bacterial cells can be administered (ingested) as they are, or they can be used as tablets, granules, powders, capsules, or powders according to general pharmaceutical manufacturing methods.

[0046] The composition for promoting IL-23 production according to an embodiment of the present invention is expected to exert its effects in all parts of the gastrointestinal tract. The gastrointestinal tract refers to the series of parts including the oral cavity, pharynx, esophagus, stomach, duodenum, small intestine, large intestine, cecum, and anus. While typical names of the gastrointestinal tract are given here as examples, the effects of ingestion (administration) of the composition for promoting IL-23 production described above can be expected to be exerted in any gastrointestinal tract with a name that is publicly known and / or falls within the scope of common general technical knowledge, such as the colon (lower part of the small intestine), ileum, jejunum (upper part of the small intestine), upper abdominal gastrointestinal tract, lower abdominal gastrointestinal tract, upper part of the large intestine, and lower part of the large intestine.

[0047] The compositions for promoting IL-23 production according to embodiments of the present invention are effective in all mammals, including humans. Therefore, various diseases can be treated or prevented by ingesting and / or administering the compositions for promoting IL-23 production. Here, mammals include all known domestic animals, pets, ornamental animals, and wild animals classified as mammals, including humans, cows, pigs, sheep, dolphins, whales, tigers, lions, cheetahs, hippos, giraffes, camels, alpacas, dogs, cats, monkeys, foxes, raccoon dogs, bears, squirrels, fur seals, sea lions, pandas, wild boars, deer, horses, orangutans, and kangaroos. Furthermore, the compositions for promoting IL-23 production according to embodiments of the present invention can also be administered and / or ingested by animals other than mammals that have a digestive tract, such as insects, reptiles, birds, fish, amphibians, and mollusks, and are expected to provide similar effects. The various diseases referred to here are not particularly limited and may be, for example, diseases that can be treated and / or prevented by inducing antimicrobial peptides. Examples of such diseases include infections caused by opportunistic bacteria, infections caused by pathogenic bacteria or fungi, Helicobacter pylori infection, bacterial translocation, persistent inflammation of the intestinal mucosa, changes in the balance of intestinal flora (such as disruption of the balance of intestinal flora), and diseases induced by these inflammations, inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, and excessive inflammation and allergic symptoms that induce differentiation of regulatory T cells. [Example]

[0048] The present invention will be described in more detail below with reference to examples, but the present invention is not limited thereto.

[0049] Example 1 1. Preparation of heat-killed lactic acid bacteria First, the following nine strains of lactic acid bacteria were prepared as examples. Lactobacillus delbrueckii subsp. bulgaricus strain 2038 Lactobacillus delbrueckii subsp. bulgaricus strain MEP201701 Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (Accession number: NITE BP-76) Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession number: FERM BP-10741) Lactobacillus gasseri strain OLL2716 (Accession number: FERM BP-6999) Lactobacillus gasseri strain OLL2809 (Accession number: NITE BP-72) Lactobacillus gasseri strain OLL2959 (NITE BP-224) Streptococcus thermophilus strain 1131 Streptococcus thermophilus strain MEP201702

[0050] Next, various Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus gasseri strains were inoculated into MRS medium, and various Streptococcus thermophilus strains were inoculated into M17 medium containing 0.5% lactose. Each medium was then placed in an AnaeroPack jar and cultured once at 37°C under an anaerobic environment. A 1% suspension of each Lactobacillus strain was then added to each medium, and the main culture was continued for 18 hours. After the main culture of each Lactobacillus strain was completed, each medium was washed with phosphate-buffered saline (PBS). The turbidity of the wash solution was measured spectrophotometrically at 650 nm, and a Lactobacillus suspension was prepared to achieve an OD of 2. Finally, each Lactobacillus suspension was incubated in a water bath at 65°C for 1 hour to prepare heat-killed Lactobacillus cells.

[0051] 2. Dendritic Cell Preparation (1) Collection of tsDCs tsDC (ECACC) were collected from the bone marrow of mice transfected with the SV40 large T antigen gene.

[0052] (2) Preparation of bone marrow-derived dendritic cells (BMDCs) from normal mice Using a cell sorter (auto MACS®, manufactured by Miltenyi Biotec), undifferentiated dendritic cells were isolated from the bone marrow of the lower limbs of 8-week-old male Balb / c mice (provided by SLC Japan). These undifferentiated dendritic cells were then cultured in RPMI medium containing granulocyte macrophage colony-stimulating factor (GM-CSF) at 37°C in a 5% CO2 environment for 8 days to obtain the desired normal mouse BMDCs.

[0053] 3. Stimulation of dendritic cells by heat-killed lactic acid bacteria 1 × 10 dendritic cells 5Dendritic cells were seeded onto a 24-well plate at 1 cell / well in a 500 μL solution. The following day, 5 μL of heat-killed lactic acid bacteria was added to each well. tsDCs were then incubated at 33°C in a 9% CO2 environment for 24 hours, and BMDCs were incubated at 37°C in a 5% CO2 environment for 24 hours. Three samples were prepared for each tsDC and BMDC. The supernatant was then collected from each culture medium. When tsDCs were used as dendritic cells, all heat-killed strains of lactic acid bacteria listed above were used. When BMDCs were used as dendritic cells, only Lactobacillus delbrueckii subsp. bulgaricus strain 2038 and Streptococcus thermophilus strain 1131 were used.

[0054] 4. Preparation of Control Samples 1 × 10 dendritic cells 5 Dendritic cells were seeded onto a 24-well plate at 1 cell / well in a volume of 500 μL. The following day, 5 μL of phosphate-buffered saline (PBS) was added to each well. tsDC dendritic cells were then cultured at 33°C under 9% CO2 for 24 hours, while BMDCs were incubated at 37°C under 5% CO2 for 24 hours. Three samples were prepared for each of tsDC dendritic cells and BMDCs. The supernatant was then collected from each culture medium.

[0055] 5. Measurement of interleukin-23 concentration in dendritic cell culture supernatant The interleukin-23 concentration in the dendritic cell culture supernatant was measured using a Mouse IL-23 Quantikine ELISA Kit (R&D Systems). Table 1 shows the results when tsDCs were used as dendritic cells, and Table 2 shows the results when BMDCs were used as dendritic cells. Figure 1 shows a bar graph of the results in Table 1, and Figure 2 shows a bar graph of the results in Table 2. As is clear from Tables 1 and 2, stimulation of dendritic cells with heat-killed lactic acid bacteria promoted interleukin-23 secretion. When tsDCs were used as dendritic cells, Lactobacillus delbrueckii subsp. bulgaricus OLL1255, Lactobacillus gasseri OLL2959, and Streptococcus thermophilus 1131 showed particularly significant effects. When BMDCs were used as dendritic cells, Streptococcus thermophilus 1131 showed particularly significant effects.

[0056] [Table 1]

[0057] [Table 2]

[0058] Example 2 1. Preparation of Heat-killed Bacteria First, as an example, the following 11 strains of lactic acid bacteria and 2 strains of bifidobacteria were prepared. Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 (Accession number: FERM BP-10741) Lactobacillus delbrueckii subsp. bulgaricus strain OLL1181 (Accession number: FERM BP-11269) Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (Accession number: NITE BP-76) Lactobacillus gasseri strain OLL2716 (Accession number: FERM BP-6999) Lactobacillus gasseri strain OLL2809 (Accession number: NITE BP-72) Lactobacillus gasseri strain MEP201801 Lactobacillus plantarum strain OLL2712 (FERM BP-11262) Lactobacillus plantarum strain MEP201802 Lactobacillus johnsonii strain MEP201803 Lactobacillus johnsonii strain OLL203565 (NITE BP-02003) Lactobacillus johnsonii strain OLL204255 (NITE BP-02004) Bifidobacterium bifidum strain OLB6378 (NITE BP-31) Bifidobacterium bifidum strain MEP201804

[0059] Next, various strains of Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus gasseri, Lactobacillus plantarum, and Lactobacillus johnsonii were inoculated into MRS medium, and various strains of Bifidobacterium bifidum were inoculated into GAM medium. Each medium was then placed in an AnaeroPack jar and cultured once at 37°C under an anaerobic environment. A 1% suspension of each strain of Lactobacillus and Bifidobacterium was then added to each medium, and the Lactobacillus and Bifidobacterium suspensions were cultured for 18 hours. After the culture was complete, each medium was washed with phosphate-buffered saline (PBS). The turbidity of the wash solution was measured spectrophotometrically at 650 nm, and Lactobacillus and Bifidobacterium suspensions were prepared to achieve an OD of 2. Finally, each lactic acid bacteria suspension and bifidobacterial suspension was incubated in a water bath at 65°C for 1 hour to prepare the target heat-killed lactic acid bacteria cells and bifidobacterial cells.

[0060] 2. Preparation of bone marrow-derived dendritic cells (BMDCs) from normal mice BMDCs from normal mice of interest were obtained according to the same procedure as described in the section "(2) Preparation of bone marrow-derived dendritic cells (BMDCs) from normal mice" in "2. Preparation of dendritic cells" in Example 1.

[0061] 3. Stimulation of dendritic cells with heat-killed bacteria Following the same procedure as described in "3. Stimulation of dendritic cells with heat-killed lactic acid bacteria" in Example 1, the above-mentioned 11 heat-killed lactic acid bacteria strains and 2 heat-killed bifidobacterial strains were added to BMDCs, and the BMDCs were incubated.

[0062] 4. Preparation of Control Samples A control sample was prepared according to the same procedure as described in "4. Preparation of Control Sample" in Example 1.

[0063] 5. Measurement of interleukin-23 concentration in dendritic cell culture supernatant The concentration of interleukin-23 in the BMDC culture supernatant was measured according to the same procedure as described in "5. Measurement of interleukin-23 concentration in dendritic cell culture supernatant" in Example 1. The results are shown in Table 3. A bar graph of the results shown in Table 3 is also shown in Figure 3. As is clear from Table 3, it was demonstrated that stimulation of BMDC with heat-killed lactic acid bacteria and heat-killed bifidobacteria promotes the secretion of interleukin-23. Here, the Lactobacillus delbrueckii subsp. bulgaricus strain OLL1255 (accession number: NITE BP-76), Lactobacillus gasseri strain OLL2809 (accession number: NITE BP-72), and Lactobacillus plantarum strain OLL2712 (FERM BP-11262) showed particularly remarkable effects.

[0064] [Table 3] [Industrial Applicability]

[0065] The composition for promoting interleukin-23 production according to the present invention can eliminate the need for troublesome steps such as changing the flavor or encapsulating the product, and can therefore be produced at low cost. [Accession number]

[0066] FERM BP-06999 FERM BP-10741 FERM BP-11262 FERM BP-11269 NITE BP-31 NITE BP-72 NITE BP-76 NITE BP-224 NITE BP-02003 NITE BP-02004

Claims

1. A composition for promoting interleukin-23 production, which contains Lactobacillus johnsonii as an active ingredient.

2. A composition for promoting interleukin-23 production, containing only Lactobacillus johnsonii as an active ingredient.

3. The Lactobacillus johnsonii strain is Lactobacillus johnsonii strain OLL203565 (accession number: NITE BP-02003) or Lactobacillus johnsonii strain OLL204255 (accession number: NITE BP-02004). The composition for promoting interleukin-23 production according to claim 1 or 2.

4. The bacteria are killed. The composition for promoting interleukin-23 production according to any one of claims 1 to 3.

5. The killed bacteria are heat-killed bacteria. The composition for promoting interleukin-23 production according to claim 4.

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