WO2018151245A1

WO2018151245A1 - Interleukin-23 production promoting composition and interleukin-23 production promoting method

Info

Publication number: WO2018151245A1
Application number: PCT/JP2018/005414
Authority: WO
Inventors: 杏輔小林; 幸夫浅見
Original assignee: 株式会社明治
Priority date: 2017-02-17
Filing date: 2018-02-16
Publication date: 2018-08-23
Also published as: JP7531261B2; JPWO2018151245A1; JP2024111238A

Abstract

The present invention provides an interleukin-23 production promoting composition that enables the elimination of cumbersome procedures such as encapsulation and flavor alteration. The interleukin-23 production promoting composition according to the present invention contains bacteria as an active ingredient. The bacteria to be used are preferably of at least one type selected from the group consisting of Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus gasseri, Streptococcus thermophilus, Lactobacillus plantarum, Lactobacillus johnsonii, and Bifidobacterium bifidum.

Description

Interleukin-23 production promoting composition and interleukin-23 production promoting method

The present invention relates to a composition for promoting production of interleukin-23, a method for promoting production of interleukin-23, and the like.

In the past, “interleukin (hereinafter abbreviated as“ IL ”)-23 secretion (production) promoter comprising a low molecular weight monocarboxylic acid such as acetic acid, lactic acid or thioglycolic acid” has been proposed ( For example, see JP 2008-127277 A).

JP 2008-127277 A

However, low molecular weight monocarboxylic acids such as acetic acid, lactic acid or thioglycolic acid have a relatively strong acidity and are difficult to take as they are. For this reason, in order to ingest a low molecular weight monocarboxylic acid without a sense of incongruity, processing such as formulation is required. Specifically, a troublesome work such as adding a sweetener or the like to change the taste or encapsulating is required.

An object of the present invention is to provide an IL-23 production promoter, a composition for promoting IL-23 production, and the like that can save troublesome work such as changing taste and encapsulating.

The composition for promoting IL-23 production according to the first aspect of the present invention contains bacterial cells as an active ingredient. That is, the bacterial cells are used as a composition for promoting IL-23 production or as a component thereof. In addition, a microbial cell may be a living microbial cell or a dead microbial cell. When the cells are dead cells, the dead cells are preferably heat-killed cells, that is, cells killed by heat. In addition, the “composition” referred to herein includes animals such as pharmaceuticals, supplements and food additives, foods and drinks (excluding animals and plants themselves), and food and drink compositions (including processed foods and drinks). Included are those that can be ingested (including humans).

As a result of intensive studies by the inventors of the present application, it has been clarified that the bacterial cells according to the first aspect of the present invention can promote the production of IL-23. That is, this microbial cell can favorably promote the production of IL-23.

Incidentally, IL-23, for example, acts on ILC3 to induce the production of IL-22. When IL-22 is recognized by small intestinal epithelial cells, expression of antibacterial peptides such as Reg3 family proteins is induced. And, microorganisms such as foreign bacteria are killed by this antibacterial peptide, and expansion of microbial infection and excessive inflammatory reaction can be suppressed. This effect is only an example, and it can be sufficiently assumed that IL-23 induces the production of other substances and other effects are manifested.

Moreover, this microbial cell is tasteless. Therefore, when preparing the composition for promoting production of IL-23, troublesome work such as changing the taste and encapsulating can be saved. Of course, the taste of the composition for promoting production of IL-23 may be changed from other viewpoints, or the composition for promoting production of IL-23 may be encapsulated.

In the above-mentioned composition for promoting IL-23 production, the microbial cells are preferably at least one of lactic acid bacteria and bifidobacteria.

In the above-mentioned composition for promoting IL-23 production, lactic acid bacteria are Lactobacillus delbrueckiispsubsp. Bulgaricus, Lactobacillus gasseri, Streptococcus thermophilus (Streptococcus). It is preferably at least one lactic acid bacterium selected from the group consisting of thermophilus, Lactobacillus plantarum and Lactobacillus johnsonii.

In the above-mentioned composition for promoting IL-23 production, lactic acid bacteria are “Lactobacillus delbrucky subspecies bulgaricus 2038, Lactobacillus delbrucky subspecies bulgaricus OLL1181 (Accession number: FERM BP- 11269), Lactobacillus delbrucky subspecies bulgaricus OLL1255 (Accession number: NITE BP-76), Lactobacillus delbrucky subspecies bulgaricus OLL1073R-1 (accession number: FERM BP-10741), lact Bacillus gasseri OLL2716 (Accession number: FERM BP-6999), Lactobacillus gaseri OLL2809 (Accession number: NITE BP-72), Lactobacillus gaseri OLL2 59 (Accession number: NITE BP-224), Lactobacillus plantarum OLL 2712 (Accession number: FERM BP-11262), Lactobacillus John Sony OLL 203565 (Accession number: NITE BP-02003), Lactobacillus John Sony OLL 204255 ( Accession number: NITE BP-02004) and at least one lactic acid bacterium selected from the group consisting of Streptococcus thermophilus 1131 ”,“ Lactobacillus delbruecki subspecies bulgaricus 2038, Lactobacillus delbrucky・ Subspecies Bulgarix OLL1181 (Accession number: FERM BP-11269), Lactobacillus delbruecki Subspecies Bull Rix OLL1255 (Accession number: NITE BP-76), Lactobacillus delbruecki Subspecies Bulgarix OLL1073R-1 (Accession number: FERM BP-10741), Lactobacillus gaseri OLLL2959 (Accession number: NITE BP-224) More preferably, it is at least one lactic acid bacterium selected from the group consisting of Lactobacillus plantarum OLL 2712 (FERM BP-11262), Lactobacillus johnson OLL 203565 (NITE BP-02003) and Streptococcus thermophilus 1131. "Lactobacillus delbruecki Subspecies Bulgarix OLL1255 (Accession number: NITE BP-76), Lactobacillus gaseri OL More preferably, it is at least one lactic acid bacterium selected from the group consisting of L2959 (Accession Number: NITE BP-224), Lactobacillus plantarum OLL2712 (FERM BP-11262), and Streptococcus thermophilus 1131.

In the composition for promoting IL-23 production, the bifidobacteria is preferably Bifidobacterium bifidum.

In addition, the aforementioned Bifidobacterium is preferably Bifidobacterium bifidum OLB6378 (accession number: NITE BP-31).

When the dendritic cells are tsDC, among the above-mentioned cells, cells that produce interleukin-23 at a concentration of 30 pg / mL or higher with respect to tsDC are preferable, and interleukins at a concentration of 50 pg / mL or higher are preferable. More preferably, microbial cells producing -23, more preferred microbial cells producing interleukin-23 at a concentration of 70 pg / mL or more, particularly preferred microbial cells producing interleukin-23 at a concentration of 90 pg / mL or more. .

Further, as described above, when the dendritic cell is tsDC, specifically, Lactobacillus delbruecki subspecies bulgaricus that produces interleukin-23 at a concentration of 30 pg / mL or more with respect to tsDC. Lactobacillus delbruecki subspecies bulgaricus producing interleukin-23 at a concentration of 40 pg / ml or greater, Lactobacillus delbruecki subspecies bulgaric producing interleukin-23 at a concentration of 90 pg / ml or greater , Streptococcus thermophilus producing interleukin-23 at a concentration of 40 pg / mL or higher, Streptococcus thermophilus producing interleukin-23 at a concentration of 70 pg / mL or higher, concentration of 60 pg / mL or higher Interleukin -23 include Streptococcus thermophilus to produce.

When the dendritic cells are BMDC, among the above-mentioned cells, cells that produce interleukin-23 at a concentration of 300 pg / mL or higher relative to BMDC are preferable, and interleukins at a concentration of 600 pg / mL or higher are preferable. More preferably, cells producing -23, more preferably cells producing interleukin-23 at a concentration of 900 pg / mL or more, more preferably cells producing interleukin-23 at a concentration of 1200 pg / mL or more. Cells that produce interleukin-23 at a concentration of 1500 pg / mL or more are more preferred, and cells that produce interleukin-23 at a concentration of 2000 pg / mL or more are particularly preferred.

In addition, as described above, when the dendritic cell is BMDC, specifically, Lactobacillus delbrukee subspecies bulgaricus that produces interleukin-23 at a concentration of 800 pg / mL or more with respect to BMDC, Lactobacillus delbruecki subspecies bulgaricus producing interleukin-23 at a concentration of 1000 pg / ml or more Lactobacillus delbruecki subspecies bulgaric producing interleukin-23 at a concentration of 1300 pg / ml or more Lactobacillus delbruecki subspecies bulgaricus producing interleukin-23 at a concentration of 1600 pg / ml or more, Streptococcus producing interleukin-23 at a concentration of 2000 pg / ml or more S. thermophilus, Lactobacillus gasseri that produces interleukin-23 at a concentration of 800 pg / mL or more, Lactobacillus gasseri that produces interleukin-23 at a concentration of 1000 pg / mL, or 1300 pg / mL or higher Lactobacillus gasseri that produces leukin-23, Lactobacillus gasseri that produces interleukin-23 at a concentration of 1600 pg / ml or more, Lactobacillus plantarum that produces interleukin-23 at a concentration of 1000 pg / ml or more, Lactobacillus plantarum producing interleukin-23 at a concentration of 1500 pg / mL or higher, Lactobacillus johnsonii producing interleukin-23 at a concentration of 300 pg / mL or higher, 800 pg / m Lactobacillus johnsonii producing interleukin-23 at the above concentration, Bifidobacterium bifidum producing interleukin-23 at a concentration of 1100 pg / ml or more, interleukin-23 at a concentration of 1300 pg / ml or more Examples include Bifidobacterium bifidum to be produced.

The method according to the second aspect of the present invention is a method of using bacterial cells as an IL-23 production promoter. That is, to provide a microbial cell for use as an IL-23 production promoter.

The method according to the third aspect of the present invention is a method of promoting IL-23 production in vivo by orally administering the above-mentioned composition for promoting IL-23 production. However, the act of treating a person is excluded. The administration period is preferably 1 week or more, more preferably 2 weeks or more, further preferably 3 weeks or more, and particularly preferably 4 weeks or more.

In the IL-23 production promotion method according to the third aspect of the present invention, the bacterial cells are brought into contact with dendritic cells. That is, the bacterial cells are brought into contact with dendritic cells to promote IL-23 production.

The use of the bacterial cell according to the fourth aspect of the present invention is an act for producing a composition for promoting the production of IL-23.

FIG. 3 is a bar graph showing IL-23 secretion amount (pg / mL) when tsDC is stimulated with various lactic acid bacteria heated dead cells. FIG. 3 is a bar graph showing IL-23 secretion amount (pg / mL) when BMDC is stimulated with various lactic acid bacteria killed by heating. FIG. 3 is a bar graph showing IL-23 secretion amount (pg / mL) when BMDC was stimulated with various lactic acid bacteria and Bifidobacterium heat-killed cells.

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 forms described below.

The composition for promoting IL-23 production according to the embodiment of the present invention contains microbial cells as an active ingredient. Here, the bacterial cell may be a live cell or a dead cell. When the cells are dead cells, the dead cells are preferably heat-killed cells. As used herein, the term “heat-dead cell” is an object after the cell is killed by heating. Note that this IL-23 production promoting composition may consist only of microbial cells or may contain other components. Here, microbial cells can include, for example, lactic acid bacteria, bifidobacteria, and the like. Lactic acid bacteria are a general term for taxonomically recognized lactic acid bacteria or all related bacteria, and there are no restrictions on bacterial species or strains. In addition, bifidobacteria is a generic term for taxonomically recognized bifidobacteria or all of the related bacteria, and there is no restriction on the bacterial species or strains. In addition, although lactic acid bacteria may be classified into plant origin and animal origin depending on their origin, the lactic acid bacteria of the present invention can be used from both plant origin and animal origin.

In the embodiment of the present invention, the lactic acid bacteria are Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus gasseri, Streptococcus thermophilus (Streptococcus ラクト thermophilus), More preferably, it is at least one lactic acid bacterium selected from the group consisting of Lactobacillus plantarum and Lactobacillus johnsonii.

When the lactic acid bacterium is Lactobacillus delbruecki subspecies bulgaricus, Lactobacillus delbrucky subspecies bulgaricus 2038, Lactobacillus delbrucky subspecies bulgaricus OLL1181 (Accession number: FERM BP- 11269), Lactobacillus delbruecki subspecies bulgaricus MEP201701, Lactobacillus delbrucky subspecies bulgaricus OLL1255 (Accession number: NITE BP-76), Lactobacillus delbruecki subspecies bulgaricus OLLG1073R -1 (accession number: FERM BP-10741), and in the case of Lactobacillus gasseri, Re-OLL 2716 (Accession number: FERM BP-6999), Lactobacillus gaselli OLL2809 (Accession number: NITE BP-72), Lactobacillus gaseri OLL2959 (Accession number: NITE BP-224), Lactobacillus gaseri MEP201801 Are preferably Streptococcus thermophilus 1131 and Streptococcus thermophilus MEP201702, and in the case of Lactobacillus plantarum, Lactobacillus plantarum OLL 2712 (Accession number: FERM BP-11262), Lactobacillus plantarum MEP201802 is preferred, and when Lactobacillus johnsonii, Kutobachirusu John Sony OLL203565 (Accession Number: NITE BP-02003), Lactobacillus John Sony OLL204255 (Accession Number: NITE BP-02004), it is preferred that the Lactobacillus John Sony MEP201803.

In the embodiment of the present invention, the Bifidobacterium is preferably Bifidobacterium bifidum.

The Bifidobacterium bifidum is preferably Bifidobacterium bifidum OLB6378 (Accession number: NITE BP-31), Bifidobacterium bifidum MEP201804.

Here, Lactobacillus delbruecki subspecies bulgaricus 2038 can be obtained by isolating it from Bulgarian yogurt LB81 (registered trademark) manufactured by Meiji Co., Ltd. by a usual method. Lactobacillus delbruecki subspecies bulgaricus MEP201701 can be obtained by isolation from fermented milk from the Republic of Bulgaria by a conventional method. In addition, Lactobacillus delbruecki Subspecies Bulgarix OLL1255 was issued on February 10, 2005 (original deposit date), and is based on the Patent Microorganism Depositary Center of the National Institute of Technology and Evaluation (Kazusa Kama, Kisarazu City, Chiba Prefecture, Japan). 2-5-8, room 122), the deposit number: NITE BP-76 has been deposited internationally under the Budapest Treaty (transferred from the original deposit to the deposit under the Budapest Treaty on April 1, 2009). In addition, Lactobacillus delbruecki Subspecies Bulgarix OLL1073R-1 was established on February 22, 1999 (date of domestic consignment), National Institute of Advanced Industrial Science and Technology, Biological Depositary Center (East 1 in Tsukuba City, Ibaraki Prefecture, Japan) No. 1 Chome No. 1 (Chuo No. 6) (later centralized in the Patent Microorganism Deposit Center of the National Institute of Technology and Evaluation (Kazusa-Kamazu 2-5-8, No. 122, Kisarazu-shi, Chiba, Japan)) BP-1074 has been deposited internationally under the Budapest Treaty (transferred from the original deposit to the deposit under the Budapest Treaty on November 29, 2006). The Lactobacillus delbruecki subspecies bulgaricus OLL 1181 was issued on July 16, 2010 at the National Institute of Advanced Industrial Science and Technology, Biological Depositary Center (1st, 1st East, 1-chome, Tsukuba, Ibaraki, Japan). ) (Later unified with the Patent Microorganisms Depositary Center for Product Evaluation Technology Infrastructure (Launched in Room 122, 2-5-8, Kazusa-Kamashita, Kisarazu City, Chiba, Japan)), the Budapest Treaty under the accession number: FERM BP-11269 Has been deposited internationally. In addition, Lactobacillus gaselli OLL2716 was dated May 24, 1999 (original deposit date), and the Institute of Biotechnology, Institute of Industrial Technology, Ministry of International Trade and Industry (1-3 Higashi 1-chome, Tsukuba, Ibaraki, Japan) Independent administrative agency, Product Evaluation Technology Foundation, Patent Microorganism Deposit Center (Centralized in Room 122, 2-5-8, Kazusa Kamashizu, Kisarazu City, Chiba, Japan), under the accession number: FERM BP-6999, based on the Budapest Treaty Deposited (transferred from the original deposit to the deposit under the Budapest Treaty on January 14, 2000). In addition, Lactobacillus gaselli OLL2809, dated February 1, 2005 (original deposit date), is the Patent Microorganisms Depositary Center for Product Evaluation Technology, Japan (2-5-8 Kazusa Kamashi, Kisarazu City, Chiba Prefecture, Japan) 122 Room No.) has been deposited internationally under the Budapest Treaty under the deposit number: NITE BP-72 (transferred from the original deposit to the deposit under the Budapest Treaty on January 18, 2006). In addition, Lactobacillus gaselli OLL2959, dated March 31, 2006 (original deposit date), National Institute for Product Evaluation Technology Patent Microorganisms Deposit Center (2-5-8 Kazusa Kamashika, Kisarazu City, Chiba Prefecture, Japan) 122 No. Room) has been deposited internationally as NITE BP-224 under the Budapest Treaty (transferred from the original deposit to the deposit under the Budapest Treaty on November 21, 2007). Lactobacillus gasseri MEP201801 can be obtained by isolation from normal feces of healthy adults by conventional methods. Streptococcus thermophilus 1131 can be obtained by isolation from Bulgarian yogurt LB81 (registered trademark) manufactured by Meiji Co., Ltd. according to a conventional method. Streptococcus thermophilus MEP201702 can be isolated and obtained from fermented milk from the Republic of Bulgaria by a conventional method. In addition, Lactobacillus plantarum OLL 2712 was issued on July 2, 2010, National Institute of Advanced Industrial Science and Technology, Biological Depositary Center (1st, 1st East, 1-chome, Tsukuba, Ibaraki, Japan) Incorporated internationally under the Budapest Treaty under the accession number: FERM BP-11262, which is centralized in the Patent Product Depositary Center of the National Institute for Product Evaluation and Technology (centralized at 2-5-8, Kazusa-Kamashita 2-5-8, Kisarazu City, Chiba, Japan) Yes. Lactobacillus plantarum MEP201802 can be obtained by isolation from raw milk from Hokkaido by a conventional method. In addition, Lactobacillus johnsonii OLL203565 was issued on February 3, 2015 at the Japan Institute for Product Evaluation Technology Patent Microorganism Depositary Center (Kazusa Kamashichi, Kisarazu City, Chiba Prefecture, 2-5-8, Room 122). Deposit number: NITE BP-02003 is deposited internationally based on the Budapest Treaty. In addition, Lactobacillus johnsonii OLL204255 was issued on February 3, 2015 in the National Institute for Product Evaluation Technology Patent Microorganism Depositary Center (Room 2-5-8, Kazusa Kamashichi, Kisarazu City, Chiba, Japan). Deposit number: NITE BP-02004 is deposited internationally based on the Budapest Treaty. In addition, Lactobacillus johnsonii MEP201803 can be obtained by isolation from a dogwood pod that inhabits the mountains of Kanagawa Prefecture by a conventional method. In addition, Bifidobacterium bifidum OLB6378 was commissioned on October 26, 2004 by the National Institute for Product Evaluation Technology Patent Microorganisms Deposit Center (2-5-8 Kazusa Kamashika, Kisarazu City, Chiba Prefecture, Japan). Number: NITE BP-31 is deposited internationally based on the Budapest Treaty. Bifidobacterium bifidum MEP201804 can be obtained by isolation from healthy adult feces by conventional methods.

The form of the cells can be used even if it is frozen or freeze-dried. Furthermore, the microbial cells may be in a form in which only the microbial cells or components other than the microbial cells (for example, cryoprotectants, lyophilization protectants, etc.) are included. For example, it may be dispersed in various media such as starch and water that have been used.

The IL-23 production promoting composition according to the embodiment of the present invention exhibits its function when taken by mammals including humans. The “intake” as used herein is not limited to the administration route as long as it enters the human body, and is realized by all known administration methods such as oral administration, tube administration, enteral administration and the like. obtain. In this case, typical examples include oral intake and enteral intake via the digestive tract, but oral intake is preferable, and intake by eating and drinking is more preferable.

The composition for promoting IL-23 production according to the embodiment of the present invention may be any microorganism as long as it contains bacteria, and the number thereof is not particularly limited, but there are 100 million or more bacteria per unit package. Preferably, 500,000 or more are present, more preferably 1 billion or more. The effect can be expected as the number of cells increases, but the upper limit is 10 trillion.

The weight per unit packaging of the IL-23 production promoting composition according to the embodiment of the present invention is not particularly limited, but when there are 100 million or more cells per gram of the IL-23 production promoting composition, The weight is preferably in the range of 10 to 500 g, more preferably in the range of 25 to 250 g, more preferably in the range of 50 to 200 g, and in the range of 75 to 150 g. Most preferably. Moreover, the above-mentioned unit packaging is not limited to unit packaging per bag, box, and container, but may be unit packaging per one time included therein or unit packaging per day. It should be noted that a plurality of days, for example, a quantity suitable for intake for one week may be packaged together, or may include a plurality of individual packages.

In the embodiment of the present invention, the composition for promoting IL-23 production is desirably ingested continuously for 3 weeks or more, preferably 5 weeks or more, more preferably 8 weeks or more. In addition, since the composition for promoting the production of IL-23 can be safely ingested, the ingestion period is not particularly limited and can be permanently continued. From the viewpoint of obtaining a sufficiently effective IL-23 production effect, the intake period is preferably 8 weeks.

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

When the IL-23 production promoting composition according to the embodiment of the present invention is used as a pharmaceutical product or a food or drink, the state of the IL-23 production promoting composition is not limited, and it is a pasty product, a spray-dried product, or a freeze-dried product. The product can be used in the form of a product, a vacuum dried product, a drum dried product, a liquid product dispersed in a medium, a diluted product diluted with a diluent, a crushed product obtained by crushing a dried product with a mill or the like.

Furthermore, the composition for promoting IL-23 production according to the embodiment of the present invention can be used as a health functional food or a food for the sick. The functional health food system is intended for not only regular foods but also foods in the form of tablets, capsules, etc., taking into account trends in Japan and overseas and the consistency with conventional food systems for specified health use. . In this system, two types of foods for specific health use (individual permission type) and functional foods (standard specification type) are defined. The composition for promoting production of IL-23 according to the embodiment of the present invention is administered to animals such as humans as special-purpose foods such as foods for specified health use or nutritional functional foods, thereby preventing various infections, for example. Is possible.

In the IL-23 production promoting composition according to the embodiment of the present invention, it is preferable to display a description of its use, efficacy, function, type of active ingredient, type of functional ingredient, ingestion method, and the like. “Indication” as used herein refers to pharmaceuticals, quasi drugs, health functional foods, foods for specified health use, functional foods for nutrition, general foods, health supplements, health foods, supplements, enteral nutrition, oral cosmetics, and feed Each should be a suitable display. In addition, the “display” here includes all displays for informing the consumer of the above description. This display only needs to be a display that allows the above-described display contents to be recalled / analogized, and may include any display regardless of the purpose of display, the display contents, the object / medium to be displayed, and the like. For example, display the above description on product packaging / containers, display the above description on product advertisements / price lists or transaction documents, or display or distribute the information, electromagnetically (such as the Internet) ) By a method.

When the product obtained by packaging the composition for promoting IL-23 production according to the embodiment of the present invention is, for example, a food or drink, the food or drink includes, for example, an indication “for IL-23 production promotion” The label “contains lactic acid bacteria that promotes the production of IL-23”, the label “for antibacterial peptide enhancement”, the label “for prevention of infectious diseases”, etc. Is preferred.

Note that the wording used for the display as described above is not limited to the above-described example, and may be a word having the same meaning as that. As such terms, for example, various terms that allow the consumer to recognize IL-23 production promotion, antibacterial peptide enhancement, infection prevention effects, and the like can be accepted.

When the composition for promoting IL-23 production according to the embodiment of the present invention is used as a food or drink, the type of the food or drink is not particularly limited. Foods and beverages include, for example, milk, processed milk, soft drinks, fermented milk, yogurt, cheese, other dairy products, bread, biscuits, crackers, pizza crusts, prepared powdered milk, liquid food, food for the sick, infant milk powder, etc. It may be food, food such as powdered milk for pregnant women and lactating women, and nutritional food. In the production of such foods and drinks, the bacterial cells that are the active ingredients of the IL-23 production promoting composition according to the embodiment of the present invention are used as they are, or mixed with other foods and drinks or food ingredients. The manufacturing method in a normal food composition can be used. Moreover, it does not specifically limit about the shape of food / beverage products, The shape of food / beverage products used normally may be sufficient. For example, any shape such as solid (including powder and granule), paste, liquid, and suspension may be used, but not limited thereto. At this time, milk drink, fermented milk, soft drink, jelly drink, tablet, and powdered food are more preferable, and yogurt is more preferable.

The cells that are active ingredients of the composition for promoting IL-23 production according to the embodiment of the present invention include water, protein, carbohydrate, lipid, vitamins, minerals, organic acid, organic base, fruit juice, and flavor. As long as it is a component contained in normal foods such as functional ingredients and food additives, it can be added without any problem. In the production of the above food and drink, as protein sources, for example, 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 -Uses proteins or protein-containing raw materials commonly used in food production, such as casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein, and other hydrolysates can do. Examples of sugar sources include processed starch (in addition to text phosphorus, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of the lipid source include animal oils such as lard and fish oil, fractionated oils thereof, hydrogenated oil and transesterified oil; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, and fractionated oils thereof. And vegetable oils such as hydrogenated oil and transesterified oil. Examples of vitamins include vitamin A, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. Examples of the functional component include oligosaccharide, glucosamine, collagen, ceramide, royal jelly, polyphenol and the like. Examples of food additives include emulsifiers, stabilizers, thickeners, gelling agents, sweeteners, acidulants, preservatives, antioxidants, pH adjusters, and colorants. Various milk-derived components such as butter, dairy minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipid, and lactose can be suitably used for the production of the food and drink according to the embodiment of the present invention. It is an example.

These components can be used in combination of two or more. The raw material may be any of natural products, processed natural products, synthetic products and / or foods containing a large amount thereof.

The bacterial cell that is the active ingredient of the composition for promoting IL-23 production according to the embodiment of the present invention can be used as a drug for preventing infection or a drug for treatment. In the case of producing such a pharmaceutical product, the bacterial cell can be used as a crushed or unground product. Moreover, the microbial cell to be used may be single or multiple types.

The amount of bacterial cells in the above pharmaceutical product can be arbitrarily determined according to the purpose and application (prophylactic agent, therapeutic agent, etc.). An example of the content may be 0.001 to 100% (w / w), particularly 0.1 to 100%, based on the total amount, but the present invention is not limited to this.

The dosage of the pharmaceutical agent containing the above bacterial cells as an active ingredient can be appropriately set in consideration of various factors such as the administration route, the age, weight, and symptoms of animals to be administered including humans. As an example of a suitable dose, 1 to 1000 mg / kg / day can be mentioned as an active ingredient, but the present invention is not limited to this. For example, when ingested for preventive purposes over a long period, the amount may be smaller than the above range. In addition, since this active ingredient has no safety problem, it can be used in a larger amount than the above range.

The above-mentioned pharmaceutical dosage form is preferably a dosage form that can be administered orally in order to allow the cells of the present invention to reach the intestine. Examples of preferable dosage forms of the pharmaceutical product according to the present invention include tablets, coated tablets, capsules, granules, powders, liquids, syrups, lozenges and the like. These various preparations are prepared according to conventional methods, such as excipients, binders, disintegrants, lubricants, coloring agents, flavoring agents, solubilizing agents, suspension agents, coating agents, etc. It can be formulated by mixing adjuvants that can be usually used in the pharmaceutical preparation technical field.

In the case of using the above-mentioned microbial cells as pharmaceuticals, for example, in the case of oral administration, the microbial cells can be administered (ingested) as they are. For example, according to the general production method of pharmaceuticals, tablets, granules, powders Can be used as a capsule or powder.

The composition for promoting the production of IL-23 according to the embodiment of the present invention can be expected to exhibit effects at all sites of the digestive tract. The digestive tract is a site including a series of oral cavity, pharynx, esophagus, stomach, duodenum, small intestine, large intestine, cecum, and anus. Here, typical names of the digestive tract are illustrated. For example, the colon (lower small intestine), ileum, jejunum (upper small intestine), upper abdominal digestive tract, lower abdominal digestive tract, large intestine, lower large intestine If the digestive tract has a name in the category of publicly known and / or common general technical knowledge, it can be expected that the effects of ingestion (administration) of the above-mentioned composition for promoting IL-23 production will be exhibited.

The composition for promoting IL-23 production according to the embodiment of the present invention is effective for all mammals including humans. Therefore, various diseases can be treated or prevented by ingesting and / or administering this IL-23 production promoting composition. Mammals as used herein include humans, cows, pigs, sheep, dolphins, whales, tigers, lions, cheetahs, hippos, giraffes, camels, alpaca, dogs, cats, monkeys, foxes, raccoons, bears, squirrels, fur seals , Sea lions, pandas, wild boars, deer, horses, orangutans, kangaroos, etc., which are all known livestock, pets, appreciation animals, wild animals and the like and classified as mammals. In addition to mammals, the composition for promoting IL-23 production according to the embodiment of the present invention is also applied to animals having a digestive tract, such as insects, reptiles, birds, fishes, amphibians, molluscs and the like. Administration and / or ingestion is also possible, and it is expected that the same effects as described above can be obtained. The various diseases referred to herein are not particularly limited, and may be any diseases that can be treated and / or prevented by inducing an antibacterial peptide, for example. Examples of such diseases include infections caused by opportunistic bacteria, infections caused by pathogenic bacteria and fungi, infections caused by Helicobacter pylori, bacterial translocation, persistent intestinal mucosal inflammation, and changes in the balance of intestinal flora (intestinal Internal flora balance), diseases induced by these inflammations, Crohn's disease, inflammatory colitis such as ulcerative colitis, excessive inflammation and allergic symptoms that induce differentiation of regulatory T cells It is done.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Example 1
1. Preparation of heat-killed lactic acid bacteria First, as an example, the following 9 strains of lactic acid bacteria were prepared.
Lactobacillus delbrueckii subsp. Bulgaricus 2038
Lactobacillus delbrueckii subsp. Bulgaricus MEP201701
・ Lactobacillus delbrueckii subsp. Bulgaricus OLL1255 (Accession number: NITE BP-76)
・ Lactobacillus delbrueckii subsp. Bulgaricus OLL1073R-1 (Accession number: FERM BP-10741)
・ Lactobacillus gasseri OLL2716 (Accession number: FERM BP-6999)
・ Lactobacillus gasseri OLL2809 (Accession number: NITE BP-72)
・ Lactobacillus gasseri OLL2959 (NITE BP-224)
Streptococcus thermophilus 1131
Streptococcus thermophilus MEP201702

Next, various Lactobacillus delbruecki subspecies bulgaricus and various Lactobacillus gasseri were inoculated in MRS medium, and each Streptococcus thermophilus was inoculated in 0.5% lactose-containing M17 medium, and then each medium. Was stored in a jar containing an anero pack, and each lactic acid bacterium was activated and cultured once at 37 ° C. in an anaerobic environment. Thereafter, 1% suspension of each lactic acid bacterium was added to each medium, and each lactic acid bacterium was subjected to main culture for 18 hours. After the main culture of each lactic acid bacterium was completed, each medium was washed with phosphate buffered saline (PBS). And the turbidity of the washing | cleaning liquid was measured with wavelength 650nm with the spectrophotometer, and lactic acid bacteria suspension was prepared so that OD might be set to 2. Finally, each lactic acid bacterium suspension was incubated in a 65 ° C. hot water bath for 1 hour to prepare a target lactic acid bacterium heat-killed cell.

2. Preparation of dendritic cells (1) Collection of tsDC tsDC (ECACC) was collected from the bone marrow of a mouse into which the SV40 large T antigen gene was introduced.

(2) Preparation of bone marrow-derived dendritic cells (BMDC) from normal mice Using a cell sorter (auto MACS (registered trademark), manufactured by Miltenyi Biotech), 8-week-old male Balb / c mice (provided by SLC, Japan) After isolating only undifferentiated dendritic cells from the lower limb bone marrow, the undifferentiated dendritic cells were isolated in RPMI medium containing granulocyte macrophage colony-stimulating factor (GM-CSF) under an environment of 5% CO _2. After culturing at 0 ° C. for 8 days, the desired normal mouse BMDC was obtained.

3. Giving stimulation to dendritic cells by heat-killed cells of lactic acid bacteria Dendritic cells were seeded in a 24-well plate so that the dendritic cells became 1 × 10 ⁵ cells / well and the liquid volume became 500 μL, and the next day, 5 μL of heat-killed lactic acid bacteria were added to each well. Thereafter, tsDC was incubated at 33 ° C. for 24 hours in a 9% CO ₂ environment, and BMDC was incubated for 24 hours at 37 ° C. in a 5% CO ₂ environment. At this time, three samples were prepared for each of tsDC and BMDC. And the supernatant was collect | recovered from each culture solution. When tsDC is used as a dendritic cell, all of the nine strains of lactic acid bacteria are used. When BMDC is used as a dendritic cell, the nine strains of lactic acid bacteria are used. Among them, only heat-killed cells of Lactobacillus delbruecki subspecies bulgaricus 2038 and Streptococcus thermophilus 1131 were used.

4). Control sample preparation
Dendritic cells were seeded in a 24-well plate so that the dendritic cells were 1 × 10 ⁵ cells / well and the liquid volume was 500 μL, and the next day, 5 μL of phosphate buffered saline (PBS) was added to each well. ) Was added. Thereafter, tsDC dendritic cells were cultured for 24 hours at 33 ° C. in a 9% CO ₂ environment, and BMDCs were incubated for 24 hours at 37 ° C. in a 5% CO ₂ environment. At this time, three samples were prepared for each of tsDC dendritic cells and BMDC. And the supernatant was collect | recovered from each culture solution.

5. Measurement of Interleukin-23 Concentration in Dendritic Cell Culture Supernatant Using Mouse IL-23 Quantikine ELISA Kit (manufactured by R & D Systems), the concentration of interleukin-23 contained in the dendritic cell culture supernatant Concentration was measured. The results when tsDC was employed as dendritic cells are shown in Table 1, and the results when BMDC were employed as dendritic cells are shown in Table 2. Further, FIG. 1 shows a bar graph of the results shown in Table 1, and FIG. 2 shows a bar graph of the results shown in Table 2. As is clear from Tables 1 and 2, it was revealed that the secretion of interleukin-23 was promoted by stimulating dendritic cells with heat-killed lactic acid bacteria. When the dendritic cells were tsDC, Lactobacillus delbruecki subspecies bulgaricus OLL1255, Lactobacillus gasseri OLL2959 and Streptococcus thermophilus 1131 showed particularly remarkable effects. When the dendritic cells are BMDC, Streptococcus thermophilus 1131 showed a particularly remarkable effect.

(Example 2)
1. Preparation of heat-killed cells First, as an example, the following 11 strains of lactic acid bacteria and 2 strains of bifidobacteria were prepared.
・ Lactobacillus delbrueckii subsp. Bulgaricus OLL1073R-1 (Accession number: FERM BP-10741)
Lactobacillus delbrueckii subsp. Bulgaricus OLL1181 (Accession Number: FERM BP-11269),
・ Lactobacillus delbrueckii subsp. Bulgaricus OLL1255 (Accession number: NITE BP-76)
・ Lactobacillus gasseri OLL2716 (Accession number: FERM BP-6999)
・ Lactobacillus gasseri OLL2809 (Accession number: NITE BP-72)
Lactobacillus gasseri MEP201801
Lactobacillus plantarum OLL 2712 (FERM BP-11262)
Lactobacillus plantarum MEP201802
Lactobacillus johnsonii MEP 201803
・ Lactobacillus johnsonii OLL203565 (NITE BP-02003)
・ Lactobacillus johnsonii OLL204255 (NITE BP-02004)
・ Bifidobacterium bifidum OLB6378 (NITE BP-31)
-Bifidobacterium bifidum MEP201804

Next, various Lactobacillus delbruecki subspecies bulgaricus, various Lactobacillus gasseri, various Lactobacillus plantarum, various Lactobacillus johnsonii in MRS medium, and various Bifidobacterium bifidum in GAM medium After inoculation, each medium was stored in a jar containing an anero pack, and each lactic acid bacterium and bifidobacteria were activated and cultured once at 37 ° C. in an anaerobic environment. Thereafter, 1% suspension of each lactic acid bacterium and bifidobacteria was added to each medium, and each lactic acid bacterium and bifidobacteria were main cultured for 18 hours. After the main culture of each lactic acid bacterium and bifidobacteria was completed, each medium was washed with phosphate buffered saline (PBS). Then, the turbidity of the washing solution was measured with a spectrophotometer at a wavelength of 650 nm, and a lactic acid bacterium suspension and a bifidobacteria suspension were prepared so that the OD was 2. Finally, each lactic acid bacterium suspension and bifidobacteria suspension were incubated in a hot water bath at 65 ° C. for 1 hour to prepare target lactic acid bacterium heated dead cells and bifidobacteria heated dead cells.

2. Preparation of bone marrow-derived dendritic cells (BMDC) of normal mice It is described in the column of “(2) Preparation of bone marrow-derived dendritic cells (BMDC) of normal mice” in “2. Preparation of dendritic cells” in Example 1. The target normal mouse BMDC was obtained according to the same procedure as described above.

3. Giving stimulation to dendritic cells by heat-killed bacterial cells According to the same procedure as described in “3. Giving stimuli to dendritic cells by heat-killed bacterial cells of lactic acid bacteria” in Example 1, Lactic acid bacteria heat-killed cells and two strains of Bifidobacterium heat-killed cells were added, and BMDC was incubated.

4). Control sample preparation
A control sample was prepared according to the same procedure as described in “4. Preparation of Control Sample” in Example 1.

5. Determination of concentration of interleukin-23 contained in supernatant of dendritic cell culture solution Procedure described in “5. Measurement of concentration of interleukin-23 contained in supernatant of dendritic cell culture solution” in Example 1 According to the same procedure as described above, the concentration of interleukin-23 contained in the BMDC culture supernatant was measured. The results are shown in Table 3. In addition, FIG. 3 shows a bar graph of the results shown in Table 3. As is apparent from Table 3, it was revealed that secretion of interleukin-23 was promoted by stimulating BMDC with lactic acid bacteria heat-killed cells and bifidobacteria heat-killed cells. Here, Lactobacillus delbrueckii subsp. Bulgaricus OLL1255 (Accession number: NITE BP-76), Lactobacillus gasseri OLL2809 (Accession number: NITE BP-72) Lactobacillus plantarum OLL 2712 (FERM BP-11262) showed a particularly remarkable effect.

The composition for promoting production of interleukin-23 according to the present invention can save troublesome work such as changing the taste and encapsulating, and can be produced at low cost.

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

Composition for promoting production of interleukin-23 containing microbial cells as an active ingredient.
The composition for promoting interleukin-23 production according to claim 1, wherein the cells are dead cells.
The composition for promoting interleukin-23 production according to claim 2, wherein the dead cells are heat-killed cells.
The composition for promoting interleukin-23 production according to any one of claims 1 to 3, wherein the microbial cells are at least one of lactic acid bacteria and bifidobacteria.
The lactic acid bacteria are Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus gasseri, Streptococcus thermophilus, Lactobacillus plantarum Lactobacillus plantarum Lactobacillus plantarum The composition for promoting interleukin-23 production according to claim 4, wherein the composition is at least one lactic acid bacterium selected from the group consisting of Lactobacillus johnsonii.
The composition for promoting interleukin-23 production according to claim 4, wherein the bifidobacteria is Bifidobacterium bifidum.
A method of using bacterial cells as an interleukin-23 production promoter.
A method for promoting the production of interleukin-23 in vivo by orally administering the composition for promoting interleukin-23 production according to any one of claims 1 to 6 (excluding the act of treating a person). ).
A method for promoting the production of interleukin-23, wherein cells are brought into contact with dendritic cells.
Bacteria for use as an interleukin-23 production promoter.
Use of bacterial cells for the production of a composition for promoting the production of interleukin-23.
A method of promoting interleukin-23 production by contacting cells with dendritic cells.