Immunostimulatory composition and immunostimulation method

Abstract

Disclosed is an immunostimulatory composition comprising: a bacterium having an immunostimulating ability; and a plant extract having the effect of enhancing the immunostimulating ability of said bacterium.

Description

Immunostimulating composition and immunostimulating method

[0001] The present invention relates to a composition for immunostimulation and a method for immunostimulation.

[0002] The immune system is a biological defense mechanism that prevents foreign substances from entering the body and detects and eliminates them once they have entered the body, and it plays a very important role in maintaining health. However, factors that weaken the immune system include lack of exercise, overwork, stress, lack of sleep, smoking, and alcohol consumption, and we are exposed to the risk of a weakened immune system every day. For this reason, technological means for activating the immune system are being developed.

[0003] As such technical means, for example, compositions capable of activating immune function through ingestion of pharmaceuticals, foods, etc. have been developed. For example, Patent Document 1 discloses an interferon (IFN) production inducer containing, as an active ingredient, lactic acid bacteria or a culture or processed product thereof that can activate plasmacytoid dendritic cells (pDCs) and induce interferon (IFN) production. Patent Document 2 discloses an immunopotentiating composition containing, as an active ingredient, lactic acid bacteria having immunopotentiating activity and an ester bond between a polyhydric alcohol and a saturated fatty acid. Patent Document 3 discloses a method for producing lactic acid bacteria having enhanced immunopotentiating activity that does not contain an ester bond between a polyhydric alcohol and a fatty acid, the method comprising contacting an immunopotentiating activity enhancing composition containing, as an active ingredient, an ester bond between a polyhydric alcohol and a fatty acid with lactic acid bacteria having immunopotentiating activity, and then removing the immunopotentiating activity enhancing composition.

[0004] Here, in order to use a composition capable of activating immune function as a pharmaceutical or food product that requires quality control, it is necessary to control the composition's ability to activate immune function by enhancing or suppressing it.

[0005] International Publication No. 2012 / 091081 JP 2016-005452 A JP 2017-085975 A

[0006] An object of the present invention is to provide a novel immunostimulatory composition.

[0007] The present inventors have discovered that certain plant extracts enhance pDC activation induced by certain bacteria with immunostimulatory activity and the associated IFN-α production, and have completed the present invention.

[0008] The present invention provides, for example, the following inventions: [1] A composition for immunostimulation, comprising a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium. [2] A method for immunostimulation, comprising administering to or ingesting to a subject a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium. [3] A method for enhancing the immunostimulatory effect induced by a bacterium having immunostimulatory ability, comprising administering to or ingesting to a subject a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium. [4] An enhancer for the immunostimulatory effect induced by a bacterium having immunostimulatory ability, comprising a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium. [5] An enhancer for the immunostimulatory effect induced by a bacterium having immunostimulatory ability, comprising a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium. [6] A composition for immunostimulating use induced by bacteria having immunostimulating ability, comprising a plant extract having an effect of enhancing the immunostimulating ability of the bacteria having immunostimulating ability. [7] Use of bacteria having immunostimulating ability and a plant extract having an effect of enhancing the immunostimulating ability of the bacteria, for producing a composition for immunostimulating use. [8] Use of a plant extract having an effect of enhancing the immunostimulating ability of the bacteria, for producing a composition for immunostimulating use induced by bacteria having immunostimulating ability. [9] Use of a plant extract having an effect of enhancing the immunostimulating ability of the bacteria, for producing a composition for immunostimulating use, to be administered or ingested in combination with the bacteria having immunostimulating ability.

[10] Use of bacteria having immunostimulating ability for producing a composition for immunostimulating use, to be administered or ingested in combination with a plant extract having an effect of enhancing the immunostimulating ability of the bacteria having immunostimulating ability.

[11] A plant extract having an effect of enhancing the immunostimulating ability of the bacteria, for use in immunostimulation induced by bacteria having immunostimulating ability.

[12] A bacterium having immunostimulatory ability for use in a therapeutic method for immunostimulation, the bacterium being administered or ingested in combination with a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium.

[13] A plant extract having an effect of enhancing the immunostimulatory ability of a bacterium having immunostimulatory ability for use in a therapeutic method for immunostimulation, the plant extract being administered or ingested in combination with the bacterium having immunostimulatory ability.

[14] Use of a bacterium having immunostimulatory ability in a non-therapeutic method for immunostimulation, wherein the bacterium is administered or ingested in combination with a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium.

[15] Use of a plant extract having an effect of enhancing the immunostimulatory ability of a bacterium having immunostimulatory ability in a non-therapeutic method for immunostimulation, wherein the bacterium is administered or ingested in combination with the bacterium having immunostimulatory ability.

[16] The composition, method, enhancer, use, plant extract, or bacterium according to any of [1] to

[15] , wherein the bacterium having immunostimulatory ability is a lactic acid bacterium and / or an acetic acid bacterium.

[17] The composition, method, enhancer, use, plant extract, or bacterium according to

[16] , wherein the lactic acid bacterium is a bacterium of the genus Lactococcus.

[18] The composition, method, enhancer, use, plant extract, or bacterium according to

[17] , wherein the bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis.

[19] The composition or use according to any one of [1], [6] to

[10] , and

[16] to

[18] , wherein the content of the immunostimulatory bacterium is 0.001% by mass or more and less than 100% by mass, relative to the total amount of the composition for immunostimulation.

[20] The composition or use according to any one of [1], [6] to

[10] , and

[16] to

[19] , wherein the content of the plant extract is 0.000001% by mass or more and 90.0% by mass or less, relative to the total amount of the composition for immunostimulation.

[21] The composition, method, enhancer, use, plant extract, or bacterium according to any one of [1] to

[20] , wherein the ratio of the content (mass%) of the immunostimulatory bacterium to the content (mass%) of the plant extract ((content of plant extract) / (content of immunostimulatory bacterium)) is 0.001 or more and 50,000 or less.

[22] The composition, method, enhancer, use, plant extract, or bacterium according to any one of [1] to

[21] , wherein the immunostimulatory activity is an ability to induce interferon-α production in immune cells.

[23] The composition, method, enhancer, use, plant extract, or bacterium according to any one of [1] to

[22] , wherein the immune cells are pDCs.

[24] The composition, method, enhancer, use, plant extract, or bacterium according to any one of [1] to

[23] , wherein the immunostimulatory effect is an effect of enhancing the induction of interferon-α production in the immune cells by 1.10-fold or more compared to an immunostimulatory composition containing only the bacterium having immunostimulatory ability.

[25] The composition, method, enhancer, use, plant extract, or bacterium according to any one of [1] to

[24] , wherein the plant extract is one or more plant extracts selected from the group consisting of pineapple extract, mangosteen extract, maca extract, fermented rice germ and soybean extract, St. John's wort extract, chrysanthemum extract, herb extract, carob extract, ginger extract, hop extract, and red ginseng extract.

[26] A composition for immunostimulation, comprising a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium.

[27] A method for producing a composition for immunostimulation, comprising adding a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium to a bacterium having immunostimulatory ability.

[0009] Hereinafter, embodiments of the present invention will be described in detail, but the present invention is not limited to the following embodiments.

[0010] According to the present invention, a novel immunostimulatory composition can be provided.

[0011] While the present invention has been described with reference to specific examples and various embodiments, it will be readily apparent to those skilled in the art that many modifications and adaptations of the embodiments described herein are possible without departing from the spirit and scope of the invention.

[0012] This application claims priority based on Japanese Patent Application No. 2024-072381, filed with the Japan Patent Office on April 26, 2024, the contents of which are incorporated herein by reference in their entirety.

[0013] In the present disclosure, the phrase "one or more selected from the group consisting of" encompasses all combinations that can be formed by one element or two or more elements that constitute the group, and may be, for example, one of the elements that constitute the group, or a combination of any two, three, four, five, six, seven, eight, nine or more elements that constitute the group.

[0014] [Composition for immunostimulation] A first embodiment of the present disclosure relates to a composition for immunostimulation. The composition for immunostimulation contains bacteria having immunostimulatory ability and a plant extract having an immunostimulatory ability enhancing effect. The composition for immunostimulation according to one embodiment contains bacteria having immunostimulatory ability, and therefore has an immunostimulatory effect.

[0015] The immunopotentiating effect (immune activation effect) refers to the activation of the innate immune system in cells or living organisms, and may be an effect of increasing or maintaining (suppressing decline in) the function of the innate immune system in cells or living organisms. The immunopotentiating effect (immune activation capability) according to one embodiment may include an immune cell activation effect (immune cell activation capability), or may include or be a dendritic cell activation effect (dendritic cell activation capability). When the immunopotentiating effect includes a dendritic cell activation effect or is a dendritic cell activation effect, the bacterium having immunopotentiating capability has dendritic cell activation capability.

[0016] The dendritic cell activation effect (dendritic cell activation ability) may be an effect of increasing the amount of cytokine expression by dendritic cells when compared to when the bacteria having immunostimulatory ability is not administered, ingested, etc. The immunostimulatory effect (immunostimulatory ability) according to one embodiment may include a plasmacytoid dendritic cell activation effect (pDC activation effect) (plasmacytoid dendritic cell activation ability (pDC activation ability)), or may be a pDC activation effect (pDC activation ability). When the immunostimulatory effect includes pDC activation effect or is pDC activation effect, the bacteria have pDC activation ability. The pDC activation effect (pDC activation ability) may be an effect of increasing (ability to increase) the amount of cytokine expression by pDC when compared to when the bacteria having immunostimulatory ability is not administered, ingested, etc.

[0017] Plasmacytoid dendritic cells, also known as plasmacytoid dendritic cells, are a type of dendritic cell that constitutes the innate immune system. pDCs are the main producers of type I interferon in the body. Type I interferon exhibits growth inhibitory activity against viruses and the like. Interferon α (IFN-α) and interferon β (IFN-β) are known as representative type I interferons, and not only these but also interferon λ (IFN-λ), a type II interferon, is thought to have immunostimulatory activity. Specific examples of pDCs include CAL-1 cells. CAL-1 cells are a human plasmacytoid dendritic cell cancer cell line established from tumor cells in a patient's peripheral blood (JP 2007-044008 A), and are deposited at the National Institute of Technology and Evaluation, Biotechnology Center, Patent Microorganisms Depositary (NPMD, Japan) under deposit number FERM BP-10914.

[0018] The immunostimulatory effect (immunostimulatory ability), dendritic cell activation effect (dendritic cell activation ability), or pDC activation effect (pDC activation ability) according to one embodiment may include an effect of promoting the production of at least one or more IFNs (IFN production-promoting ability) selected from the group consisting of interferon α (IFN-α), interferon β (IFN-β), and interferon λ (IFN-λ), or may include an interferon α (IFN-α) production-promoting effect (IFN-α production-promoting ability), or may be an IFN-α production-promoting effect (IFN-α production-promoting ability).

[0019] Whether or not an immunostimulating composition according to one embodiment has an immunostimulating effect can be determined as appropriate. For example, if the function of immune cells is enhanced when cultured in the presence of an immunostimulating composition according to one embodiment, compared to when cultured in the absence of the composition, the composition is determined to have an immunostimulating effect. Furthermore, if the function of immune cells is maintained (inhibited from decline) when cultured in the presence of an immunostimulating composition according to one embodiment and a substance that reduces immune cell function, compared to when cultured in the absence of the composition and in the presence of a substance that reduces immune cell function, the composition is determined to have an immunostimulating effect. The function of immune cells may be evaluated, for example, using an activation marker molecule (protein, etc.) that indicates that the immune cells are activated, or a molecule (protein, etc.; e.g., IFN, etc.) produced when the immune cells are activated as an index.

[0020] As a more detailed example, whether or not the immunostimulating composition according to one embodiment has an immunostimulating effect can be confirmed by measuring the IFN-α concentration by the following procedures (i) to (iv): (i) Mouse-derived bone marrow cells from which red blood cells have been removed are added to RPMI medium prepared so that each component described in the <Composition of Medium> below has the respective final concentration described in the <Composition of Medium> below at a concentration of 1 x 10 ６ <Culture medium composition> 10% by volume FBS 100 U / mL penicillin / streptomycin 1 mM sodium pyruvate 2.5 mM HEPES 1% by mass non-essential amino acids for MEM (NEAA) 50 μM β-mercaptoethanol 100 ng / mL Flt-3L (ii) 1 mL of the prepared cell suspension was seeded on a plate, and the plate was then heated in a CO ２ Incubated at 37°C and 5% CO ２ (iii) The bone marrow cells containing the induced pDCs are cultured at 2 × 10 ５The bacteria are suspended at a concentration of 1 mg / mL, and 200 μL of the suspension is seeded onto a 96-well plate, to which 2 μL of a composition containing the bacteria adjusted to a concentration of 1 mg / mL with PBS is added. (iv) After 24 hours, the culture supernatant is collected, and the IFN-α concentration is measured by ELISA using an IFN-α measurement kit.

[0021] As used herein, "bacteria having immunostimulatory activity" refers to bacteria that have the ability to activate (activate) immune cells. "Immunostimulatory activity" may refer to any ability to activate immune cells, such as the ability to induce IFN production in immune cells, the ability to induce production of at least one IFN selected from the group consisting of IFN-α, IFN-β, and IFN-λ in immune cells, or the ability to induce IFN-α production in immune cells. "Immunostimulatory activity" may be either live or killed bacteria, or may be a disrupted product of live or killed bacteria, a lyophilized product of live or killed bacteria, a disrupted product of the lyophilized product, a culture medium, an extract of the culture medium, or a processed product thereof. Here, examples of the processed product include those treated with an enzyme treatment, heat treatment, or the like, or those recovered by ethanol precipitation of the treated product. The "immunostimulatory bacteria" are preferably killed bacteria, and more preferably crushed killed bacteria, freeze-dried products, or crushed freeze-dried products that have been heat-treated (heat-killed bacteria). When the "immunostimulatory bacteria" are killed bacteria, they can be produced by killing live bacteria by heat treatment, pressurization, high-pressure steam treatment, electromagnetic wave treatment, electron beam treatment, radiation treatment, ultraviolet treatment, alcohol treatment, electrolyzed water treatment, or the like, and then drying them as needed by freeze-drying, spray drying, drum drying, hot air drying, vacuum drying, or the like.

[0022] Whether or not the bacteria according to this embodiment have immunostimulatory activity is not particularly limited, and the bacteria may be determined to have immunostimulatory activity if the function of immune cells is enhanced when the immune cells are cultured in the presence of the bacteria having immunostimulatory activity compared to when the immune cells are cultured in the absence of the bacteria. As a more detailed example, whether or not the bacteria according to this embodiment have immunostimulatory activity can be confirmed by measuring the IFN-α concentration according to the above steps (i) to (iv).

[0023] Whether the bacteria according to this embodiment have immunostimulatory activity or not was determined by comparing the bacteria at a final concentration of 10 μg / mL with a final concentration of 2 × 10 cells containing pDCs obtained by culturing the bacteria collected from mouse bone marrow in a cell culture medium containing 100 ng / mL Flt3-L for 7 days. ５ By co-culturing the cells with bone marrow cells at a concentration of 1000 cells / mL for 24 hours, the IFN-α produced can be expressed as an index of IFN-α production of 30 pg / mL or more, preferably 50 pg / mL or more, more preferably 60 pg / mL or more, more preferably 70 pg / mL or more, more preferably 80 pg / mL or more, more preferably 90 pg / mL or more, more preferably 100 pg / mL or more, more preferably 150 pg / mL or more, more preferably 200 pg / mL or more, more preferably 250 pg / mL or more, more preferably 300 pg / mL or more, more preferably 400 pg / mL or more, more preferably 500 pg / mL or more, more preferably 600 pg / mL or more, more preferably 700 pg / mL or more, and particularly preferably 800 pg / mL or more.

[0024] The bacteria according to this embodiment are not particularly limited, and may be bacteria that are harmless to the human body. In one embodiment, the bacteria may be gram-positive or gram-negative bacteria. The bacteria according to this embodiment may be lactic acid bacteria and / or acetic acid bacteria, and particularly lactic acid bacteria. The bacteria having immunostimulatory ability according to this embodiment may be one type of bacteria or a mixture of two or more types of bacteria, such as one or more types of lactic acid bacteria, one or more types of acetic acid bacteria, or a mixture of one or more types of lactic acid bacteria and one or more types of acetic acid bacteria.

[0025] Lactic acid bacteria are bacteria that produce lactic acid as a metabolite. Examples of lactic acid bacteria include bacteria of the genus Oenococcus, Bifidobacterium, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, and Pseudomonas. Examples of such bacteria include bacteria of the genus Pediococcus, Streptococcus, Enterococcus, Lactobacillus, Bacillus, and Heyndrickxia (formerly classified as Bacillus).

[0026] In addition, the Lactobacillus bacteria in the present invention includes bacteria that were classified into the Lactobacillus genus before the reclassification of the Lactobacillus genus.For example, with the reclassification of the Lactobacillus genus, the genus Acetylactobacillus, the genus Agrilactobacillus, the genus Amylolactobacillus, the genus Apilactobacillus, the genus Bombilactobacillus, the genus Compan ... The genus Lactobacillus, the genus Dellaglioa, the genus Fructilactobacillus, the genus Furfurilactobacillus, the genus Holzapfelia, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Lapidilactob ... The genus Lapidilactobacillus, the genus Latilactobacillus, the genus Lentilactobacillus, the genus Levilactobacillus, the genus Ligilactobacillus, the genus Limosilactobacillus, the genus Liquorilactobacillus, The present invention includes bacteria classified into the genera Lactobacillus, Loigolactobacillus, Paralactobacillus, Paucilactobacillus, Schleiferilactobacillus, and Secundilactobacillus.

[0027] Among the above, examples of bacteria include bacteria of the genus Oenococcus, bacteria of the genus Bifidobacterium, bacteria of the genus Lentilactobacillus, bacteria of the genus Weissella, bacteria of the genus Tetragenococcus, bacteria of the genus Lactococcus, bacteria of the genus Leuconostoc, bacteria of the genus Pediococcus, bacteria of the genus Pseudomonas aeruginosa ... Bacteria of the genus Lactococcus, Enterococcus, Lactobacillus, Lactiplantibacillus, Lacticaseibacillus, and Limosilactobacillus are preferred, and bacteria of the genus Lactococcus are more preferred.

[0028] The above-mentioned Oenococcus bacteria are not particularly limited, but examples thereof include Oenococcus oeni, etc. Specific examples of Oenococcus bacteria include Oenococcus oeni JCM6125, etc.

[0029] The Bifidobacterium genus is not particularly limited, but examples thereof include Bifidobacterium animalis subsp. lactis and Bifidobacterium longum subsp. infantis. Specific examples of Bifidobacterium include Bifidobacterium animalis subsp. lactis JCM10602, Bifidobacterium longum subsp. infantis JCM1222, and Bifidobacterium longum subsp. longum BB536.

[0030] The above-mentioned Weissella genus bacteria is not particularly limited, but examples thereof include Weissella paramesenteroides and Weissella viridescens. Specific examples of Weissella genus bacteria include Weissella paramesenteroides JCM9890 and Weissella viridescens JCM1174.

[0031] The above-mentioned Tetragenococcus bacteria are not particularly limited, but include, for example, Tetragenococcus halophilus, etc. Specific examples of Tetragenococcus bacteria include Tetragenococcus halophilus NRIC0098, Tetragenococcus halophilus No. 1, etc.

[0032] The Lactococcus bacteria are not particularly limited, and examples thereof include Lactococcus lactis, Lactococcus lactis subsp. lactis, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. hordniae, and Lactococcus culbatus. Examples of suitable strains of Lactococcus include Lactococcus curvatus and Lactococcus plantarum.

[0033] Specific examples of the Lactococcus bacteria include Lactococcus lactis subsp. lactis JCM5805 (hereinafter sometimes referred to as JCM5805), Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NRIC1150, Lactococcus lactis subsp. lactis JCM20101, Lactococcus lactis subsp. lactis JCM7638, Lactococcus lactis subsp. lactis ATCC 7963, Lactococcus lactis subsp. lactis ATCC 7962, Lactococcus lactis subsp. lactis ATCC 29146, and Lactococcus lactis subsp. lactis ATCC lactis ATCC 19435, Lactococcus lactis subsp. lactis ATCC 15577, Lactococcus lactis subsp. lactis ATCC 15346, Lactococcus lactis subsp. lactis ATCC 13675, Lactococcus lactis subsp. lactis ATCC 12929, Lactococcus lactis subsp. lactis ATCC 11955, Lactococcus lactis subsp. lactis ATCC 11454, Lactococcus lactis subsp. lactis ATCC 11007, Lactococcus garvieae NBRC100934, Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp. cremoris NBRC100676, Lactococcus lactis subsp. holdoniae JCM1180, Lactococcus lactis subsp. holdoniae JCM11040, Lactococcus culbatus JCM1096, and Lactococcus plantarum JCM11056.

[0034] The Leuconostoc genus bacteria is not particularly limited, but examples thereof include Leuconostoc carnosum and Leuconostoc lactis. Specific examples of Leuconostoc genus bacteria include Leuconostoc carnosum JCM9695 and Leuconostoc lactis NBRC12455.

[0035] The above-mentioned Pediococcus bacteria are not particularly limited, but examples thereof include Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus cellicola, Pediococcus claussenii, Pediococcus damnosus, and Pediococcus ethanolidurans. ethanolidurans), Pediococcus inopinatus, Pediococcus parvulus, Pediococcus stillesii, etc. Specific examples of the genus Pediococcus include Pediococcus acidilactici JCM8797, Pediococcus acidilactici K15, and Pediococcus damnosus JCM5886, etc.

[0036] The Streptococcus bacteria are not particularly limited, but include, for example, Streptococcus thermophilus, etc. Specific examples of Pediococcus bacteria include, for example, Streptococcus thermophilus SBC8781, etc.

[0037] The Enterococcus bacteria are not particularly limited, but examples thereof include Enterococcus alcedinis, Enterococcus faecalis, etc. Specific examples of Enterococcus bacteria include Enterococcus faecalis EC-12, etc.

[0038] The Lactobacillus bacteria are not particularly limited, but examples thereof include Lactobacillus paracasei, Lactobacillus paracasei subsp. paracasei, Lactobacillus paracasei subsp. paracasei, Lacticaseibacillus paracasei subsp. paracasei, Lactobacillus delbrueckii, Lactobacillus acidophilus, and the like. Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus rhamnosus, Lacticaseibacillus rhamnosus, Lactobacillus gasseri, Lactobacillus acidophilus acidophilus), Lactobacillus bulgaricus, Lactobacillus parakefiri, Lentilactobacillus parakefiri, Lactobacillus plantarum, Lactiplantibacillus plantarum, Lactobacillus plantarum subsp. plantarum plantarum), Lactiplantibacillus plantarum subsp. plantarum,Lactobacillus pentosus, Lactiplantibacillus pentosus, Lactobacillus helveticus, Limosilactobacillus reuteri subsp. reuteri, Lactobacillus crispatus, Lactobacillus paraplantarum paraplantarum), Lactiplantibacillus paraplantarum, and Lactobacillus johnsonii.

[0039] Specific examples of Lactobacillus bacteria include Lactobacillus paracasei KW3110, Lactobacillus paracasei MCC1849, Lactobacillus paracasei K71, Lactobacillus paracasei K-2, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505, Lactobacillus gasseri SBT2055, Lactobacillus acidophilus L-92, Lactobacillus casei subsp. casei 327, Lactobacillus (newly classified as Lacticaseibacillus) casei Shirota, Lactobacillus bulgaricus OLL1073R-1, Lactobacillus parakeefili (newly classified as Lentilactobacillus parakeefili) JCM8573, Lactobacillus plantarum (newly classified as Lactipranchibacillus plantarum) L-137, Lactobacillus pentosus (newly classified as Lactipranchibacillus pentosus) ONRICb0240, Lactobacillus paraplantarum (newly classified as Lactipranchibacillus paraplantarum) JCM1149, Lactobacillus acidophilus JCM1021, Lactobacillus acidophilus JCM1132, Lactobacillus helveticus JCM1003, rimosyl Lactobacillus reuteri subsp. reuteri JCM1112, Lactobacillus crispatus JCM1185, and Lactobacillus johnsonii JCM2012.

[0040] The Bacillus bacteria are not particularly limited, but include, for example, Bacillus coagulans, etc. Specific examples of Bacillus bacteria include Bacillus coagulans SANK70258 strain, etc.

[0041] Examples of bacteria of the genus Hendricksia include, but are not limited to, Hendrickxia coagulans (also known as Bacillus coagulans). Specific examples of bacteria of the genus Hendrickxia include Hendrickxia (Bacillus) coagulans SANK 70258 and Hendrickxia (Bacillus) coagulans BC99.

[0042] Acetic acid bacteria are bacteria that produce acetic acid as a metabolite. Examples of acetic acid bacteria include, but are not limited to, bacteria of the genus Gluconacetobacter, Acetobacter, and Gluconobacter, preferably bacteria of the genus Gluconacetobacter, more preferably Gluconacetobacter hansenii, and even more preferably Gluconacetobacter hansenii GK-1.

[0043] In addition to the above, the lactic acid bacteria may be bacteria of the genus Akkermansia, Bacteroides, Blautia, Clostridium, Collinsella, Faecalibacterium, Faecalicatena, Lacrimispora, Paeniclostridium, Parabacteroides, or Roseburia.

[0044] Specific examples of Akkermansia include Akkermansia muciniphila JCM30893.

[0045] Specific examples of bacteria of the genus Bacteroides include Bacteroides caccae JCM9498T, Bacteroides fragilis JCM11019T, Bacteroides fragilis JCM11017, Bacteroides fragilis JCM17586, Bacteroides fragilis JCM17587, Bacteroides ovatus JCM5824T, Bacteroides setaiotaomicron ATCC29148T, Bacteroides setaiotaomicron ATCC29741, Bacteroides setaiotaomicron ATCC12290, Bacteroides uniformis JCM5828T, and Bacteroides uniformis JCM13286, Bacteroides uniformis JCM13287 and Bacteroides uniformis JCM13288.

[0046] Specific examples of the genus Blautia include Blautia acetiggens JCM34803T, Blautia ammoniilytica JCM34802T, Blautia algi JCM31394T, Blautia caekimuris JCM34498T, Blautia coccoides JCM1395T, Blautia faeces JCM17205T, Blautia glucellacea JCM17039T, Blautia hansenii JCM14655, Blautia hansenii JCM35484, Blautia hominis JCM32276T, Blautia hydrogenotrophica JCM31266, and Blautia liqualis. JCM34225T, Blautia luti JCM17040T, Blautia obeum JCM31340, Blautia producta JCM1471T, Blautia pseudococcoides JCM35243T, Blautia shinkii JCM14657T, Blautia wechslerae JCM31267 and Blautia wechslerae JCM35486.

[0047] Specific examples of the genus Clostridium include Clostridium butyricum JCMNT, Clostridium nexile JCM31500T, and Clostridium symbiosum JCM1297T.

[0048] Specific examples of Collinsella bacteria include Collinsella aerofaciens JCM10188T, Collinsella intestinalis JCM10643T, Collinsella stercoris JCM10641T, and Collinsella tanakaei JCM16071T.

[0049] Specific examples of the genus Faecalibacterium include Faecalibacterium hattorii JCM39210, Faecalibacterium longum JCM39208, Faecalibacterium prausnitzii JCM31915, Faecalibacterium prausnitzii JCM39207, and Faecalibacterium prausnitzii JCM39209.

[0050] Specific examples of the genus Faecalicatena include Faecalicatena oroticum JCM1429T.

[0051] Specific examples of the genus Lacrimispora include Lacrimispora celerecrescens JCM15734T, Lacrimispora sphenoides JCM1415T, and Lacrimispora xylanolytica JCM15735T.

[0052] Specific examples of the genus Paeniclostridium include Paeniclostridium sordellii JCM3814T.

[0053] Specific examples of bacteria of the genus Parabacteroides include Parabacteroides meldae JCM9497T.

[0054] Specific examples of the genus Roseburia include Roseburia hominis JCM17582, Roseburia intestinalis JCM17583, and Roseburia inulinovorans JCM17584.

[0055] In one preferred embodiment, the bacterium is Lactobacillus rhamnosus CRL1505 or a mutant thereof, Gluconacetobacter hansenii or a mutant thereof, Lactobacillus acidophilus (GK-1, Lactobacillus acidophilus) L-92 or a mutant thereof, Lactobacillus plantarum L-137 or a mutant thereof, or Lactobacillus bulgaricus. The bacterium may be at least one bacterium selected from the group consisting of Lactococcus lactis subsp. lactis JCM5805 or a mutant strain thereof, and Lactococcus lactis subsp. lactis JCM5805 or a mutant strain thereof.

[0056] Of the above-mentioned lactic acid bacteria and acetic acid bacteria, the JCM strain can be obtained from the Microbial Materials Development Laboratory, BioResource Center, RIKEN (1-1 Takanodai 3-chome, Tsukuba, Ibaraki Prefecture), the NBRC strain can be obtained from the Biological Genetic Resources Division, National Institute of Technology and Evaluation (5-8 Kazusa Kamatari 2-chome, Kisarazu, Chiba Prefecture), the NRIC strain can be obtained from the Tokyo University of Agriculture and Technology Culture Collection (1-1 Sakuragaoka 1-chome, Setagaya-ku, Tokyo), and the ATCC strain can be obtained from the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia, USA). In addition to being obtained from public institutions, the above-mentioned lactic acid bacteria and acetic acid bacteria can also be obtained by isolation or purification using known methods from commercially available products containing lactic acid bacteria or acetic acid bacteria.

[0057] As mentioned above, the JCM strain can be obtained from the Microbial Materials Development Laboratory of the RIKEN BioResource Center, but the present invention can also use the same strain of the JCM strain stored in a collection institution other than the RIKEN BioResource Center Microbial Materials Development Laboratory. Specifically, the same strain of Lactococcus lactis subsp. lactis JCM 5805 can be obtained from the National Institute of Technology and Evaluation, Biological Genetic Resources Division (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture), the Tokyo University of Agriculture, Strain Collection (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia, USA), etc. Lactococcus lactis subsp. lactis JCM 5805 has been deposited with the American Type Culture Collection as Lactococcus lactis subsp. lactis ATCC 9936 and Lactococcus lactis subsp. lactis ATCC 19435.

[0058] The mutant strain may be any strain having properties (e.g., immunostimulatory ability) that can achieve the object of the present technology. Furthermore, the mutant strain is preferably a bacterium that has the same bacteriological properties as the above-mentioned bacterial strain and has immunostimulatory ability equivalent to or greater than that of the above-mentioned bacterial strain. Whether a mutant strain has immunostimulatory ability equivalent to or greater than that of the above-mentioned bacterial strain can be confirmed, for example, by the evaluation method described below or the method described in the Examples below.

[0059] Such mutant strains may be constructed by non-artificially introducing mutations into the strains. Alternatively, such mutant strains may be constructed by artificially introducing mutations into the strains, for example, by introducing mutations into the bacterium by treatment with a mutagen such as ultraviolet (UV) or a DNA alkylating agent, or by introducing mutations into the strains by known genetic engineering methods such as gene recombination or gene editing typified by CRISPR-Cas9.

[0060] The bacteria according to this embodiment can be cultured by a conventional method depending on the species. For example, when the bacteria are lactic acid bacteria or acetic acid bacteria, they can be cultured by a conventional method using a conventional medium. The medium is not particularly limited as long as it is capable of growing the bacteria. For example, a medium prepared by adding an appropriate plant extract to MRS medium, GAM medium, or M17 medium can be used, and inorganic salts, vitamins, and / or amino acids, etc., may be added as appropriate. Culturing can be carried out at 25 to 40°C for several hours to several days.

[0061] The effective amount of the bacteria contained in the composition of the present embodiment is not particularly limited as long as it is an amount that can exert immunostimulatory activity, and may vary depending on the subject of application. When the subject of application is a mammal such as a human, the lower limit is, for example, 1 × 10 ８ pieces, 1×10 ９ pcs or 1 x 10 １０ The upper limit can be set to, for example, 1×10 １４ pieces, 1×10 １３ pieces, 1×10 １２ These upper and lower limits can be arbitrarily combined, and the intake range can be, for example, 1 × 10 ８ 1x10 or more １４ Less than or equal to 1 x 10 ８ 1x10 or more １３ Less than or equal to 1 x 10 ８ 1x10 or more １２ Less than or equal to 1 x 10 ９ 1x10 or more １４ Less than or equal to 1 x 10 ９ 1x10 or more １３ Less than or equal to 1 x 10 １０ 1x10 or more １４ Less than or equal to 1 x 10 １０ 1x10 or more １３ 10 or less, or 1 x 10 １０ 1x10 or more １２The bacterial cell count can be measured using a known microscope, flow cytometer, or non-culture rapid microorganism testing device (e.g., ELESTA PixeeMo (AFI Technology Co., Ltd.)), but measurement using a microscope is preferred from the viewpoint of high versatility. The composition having immunopotentiating ability may be used in the form of, for example, a food composition, a pharmaceutical composition, a quasi-drug, a bacterial bulk powder (a powder of dried bacterial cells or a powder containing the same), or a feed.

[0062] The content of bacteria having immunostimulatory ability in the immunostimulatory composition of one embodiment is not particularly limited as long as it is an amount that satisfies the effective amount of immunostimulation, and therefore may vary depending on the form of the immunostimulatory composition of this embodiment. For example, the dry mass of the bacterium according to this embodiment relative to the total mass of the immunostimulating composition of this embodiment may be 0.0001% by mass or more, 0.001% by mass or more, 0.005% by mass or more, 0.01% by mass or more, 0.02% by mass or more, 0.05% by mass or more, 0.10% by mass or more, 0.30% by mass or more, 1.0% by mass or more, 1.5% by mass or more, 2.0% by mass or more, 2.5% by mass or more, 3.0% by mass or more, 5.0% by mass or more, 7.0% by mass or more, or 10.0% by mass or more, or may be less than 100% by mass, 95% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 30% by mass or less, 25% by mass or less, 20% by mass or less, or 15% by mass or less.

[0063] These lower limit values and upper limit values can be arbitrarily combined with each other. For example, with respect to the total mass of the immunostimulating composition of the present embodiment, the dry mass of the bacteria according to the present embodiment is 0.0001% by mass or more and less than 100% by mass, 0.0001% by mass or more and 95% by mass or less, 0.0001% by mass or more and 90% by mass or less, 0.0001% by mass or more and 80% by mass or less, 0.0001% by mass or more and 70% by mass or less, 0.0001% by mass or more and 60% by mass or less, 0.0001% by mass or more and 5% by mass or less, 0.0001% by mass or more and 30% by mass or less, 0.0001% by mass or more and 25% by mass or less, 0.0001% by mass or more and 20% by mass or less, 0.0001% by mass or more and 15% by mass or less, 0.001% by mass or more and less than 100% by mass, 0.001% by mass or more and 95% by mass or less, 0.001% by mass or more and 90% by mass or less, 0.001% by mass or more and 80% by mass or less, 0.001% by mass or more and 70% by mass or less, 0.001% by mass or more and 60% by mass or less, 0.001% by mass or more and 50% by mass or less, 0.001% by mass or more and 30% by mass or less, 0.001% by mass or more and 25% by mass or less, 0.001% by mass or more and 20% by mass or less, 0.001% by mass or more and 15% by mass or less, 0.005% by mass or more and less than 100% by mass, 0.005% by mass or more and 95% by mass or less, 0.005% by mass or more and 90% by mass or less, 0.005% by mass or more and 80% by mass or less, 0.005% by mass or more and 70% by mass or less, 0.005% by mass or more and 60% by mass or less, 0.005% by mass or more and 50% by mass or less, 0.005% by mass or more and 30% by mass or less, 0.005% by mass or more and 25% by mass or less, 0.005% by mass or more and 20% by mass or less, 0.005% by mass or more and 15% by mass or less, 0.01% by mass or more and less than 100% by mass, 0.01% by mass or more and 95% by mass or less, 0.01% by mass or more and 90% by mass or less, 0.01% by mass or more and 80% by mass or less, 0.01% by mass or more and 70% by mass or less, 0.01% by mass or more and 60% by mass or less, 0.01% by mass or more and 50% by mass or less, 0.01% by mass or more and 30% by mass or less, 0.01% by mass or more and 25% by mass or less, 0.01% by mass or more and 20% by mass or less, 0.01% by mass or more and 15% by mass or less, 0.02% by mass or more and less than 100% by mass, 0.02% by mass or more and 95% by mass or less, 0.02% by mass or more and 90% by mass or less, 0.02% by mass or more and 80% by mass or less, 0.02% by mass or more and 70% by mass or less, 0.02% by mass or more and 60% by mass or less, 0.02% by mass or more and 50% by mass or less,0.02% to 30% mass, 0.02% to 25% mass, 0.02% to 20% mass, 0.02% to 15% mass, 0.05% to 100% mass (not fully expressed), 0.05% to 95% mass, 0.05% to 90% mass, 0.05% to 80% mass, 0.05% to 70% mass, 0.05% to 60% mass, 0.05% to 50% mass, 0.05% to 30% mass, 0.05% to 25% mass, 0.05% to 20% mass, 0.05% mass The following are categories of product weight percentages: 15% or less, 0.10% or more but less than 100%, 0.10% or more but less than 95%, 0.10% or more but less than 90%, 0.10% or more but less than 80%, 0.10% or more but less than 70%, 0.10% or more but less than 60%, 0.10% or more but less than 50%, 0.10% or more but less than 30%, 0.10% or more but less than 25%, 0.10% or more but less than 20%, 0.10% or more but less than 15%, 0.30% or more but less than 100%, 0.30% or more but less than 95%, and 0.30% or more but less than 90%. Below, 0.30% mass% to 80% mass%, 0.30% mass% to 70% mass%, 0.30% mass% to 60% mass%, 0.30% mass% to 50% mass%, 0.30% mass% to 30% mass%, 0.30% mass% to 25% mass%, 0.30% mass% to 20% mass%, 0.30% mass% to 15% mass%, 1.0% mass% to 100% mass%, 1.0% mass% to 95% mass%, 1.0% mass% to 90% mass%, 1.0% mass% to 80% mass%, 1.0% mass% to 70% mass%, 1.0% mass% to 60% mass%, 1.0% mass% to 50% mass. Below 1%, 1.0% to 30%, 1.0% to 25%, 1.0% to 20%, 1.0% to 15%, 1.5% to 100%, 1.5% to 95%, 1.5% to 90%, 1.5% to 80%, 1.5% to 70%, 1.5% to 60%, 1.5% to 50%, 1.5% to 30%, 1.5% to 25%, 1.5% to 20%, 1.5% to 15%.2.0% or more of quality, less than 100% of quality; 2.0% or more of quality, less than 95% of quality; 2.0% or more of quality, less than 90% of quality; 2.0% or more of quality, less than 80% of quality; 2.0% or more of quality, less than 70% of quality; 2.0% or more of quality, less than 60% of quality; 2.0% or more of quality, less than 50% of quality; 2.0% or more of quality, less than 30% of quality; 2.0% or more of quality, less than 25% of quality; 2.0% or more of quality, less than 20% of quality; 2.0% or more of quality, less than 15% of quality; 2.5% or more of quality, less than 100% of quality; 2.5% or more of quality, less than 95% of quality; 2.5% or more of quality, less than 90% of quality; 2.5% or more of quality, less than 80% of quality; 2.5% of quality... 2.5% to 70% mass, 2.5% to 60% mass, 2.5% to 50% mass, 2.5% to 30% mass, 2.5% to 25% mass, 2.5% to 20% mass, 2.5% to 15% mass, 3.0% to 100% mass, 3.0% to 95% mass, 3.0% to 90% mass, 3.0% to 80% mass, 3.0% to 70% mass, 3.0% to 60% mass, 3.0% to 50% mass, 3.0% to 30% mass, 3.0% to 25% mass. Less than 100% of the quantity, 3.0% to 20% of the quantity, 3.0% to 15% of the quantity, 5.0% to 100% of the quantity, less than 95% of the quantity, 5.0% to 90% of the quantity, 5.0% to 80% of the quantity, 5.0% to 70% of the quantity, 5.0% to 60% of the quantity, 5.0% to 50% of the quantity, 5.0% to 30% of the quantity, 5.0% to 25% of the quantity, 5.0% to 20% of the quantity, 5.0% to 15% of the quantity, 7.0% to 100% of the quantity, less than 95% of the quantity, 5.0% to 95% of the quantity. 7.0% quality or higher, below 90% quality; 7.0% quality or higher, below 80% quality; 7.0% quality or higher, below 70% quality; 7.0% quality or higher, below 60% quality; 7.0% quality or higher, below 50% quality; 7.0% quality or higher, below 30% quality; 7.0% quality or higher, below 25% quality; 7.0% quality or higher, below 20% quality; 7.0% quality or higher, below 15% quality; 10.0% quality or higher, below 100% quality; 10.0% quality or higher, below 95% quality; 10.0% quality or higher, below 90% quality; 10.0% quality or higher, below 80% quality; 10.0% quality or higher, below 70% quality; 10.0% quality or higher, below 60% quality.The dry mass of the bacterium according to this embodiment may be 10.0% by mass or more and 50% by mass or less, 10.0% by mass or more and 30% by mass or less, 10.0% by mass or more and 25% by mass or less, 10.0% by mass or more and 20% by mass or less, or 10.0% by mass or more and 15% by mass or less. The above-mentioned dry mass of the bacterium according to this embodiment can be used as a daily dose for an adult weighing 60 kg, for example. The dose can be set depending on factors such as the health condition of the person receiving or ingesting the bacterium, the method of administration or ingestion, and the combination with other agents.

[0064] The number of bacteria having immunostimulatory ability in the immunostimulatory composition according to this embodiment is 1.0 × 10 ３ Cells / mL or more, 1.0 x 10 ４ Cells / mL or more, 1.0 x 10 ５ Cells / mL or more, 1.0 x 10 ６ Cells / mL or more, 1.0 x 10 ７ cells / mL or 4.0 x 10 ７ cells / mL or more, and may be 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 １０ Cells / mL or less, 3.0 x 10 ９ cells / mL or less or 1.0 x 10 ９ These lower and upper limits can be combined in any combination. For example, the number of bacteria having immunostimulatory ability in the immunostimulatory composition according to this embodiment may be 1.0 × 10 ３ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 ３ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 1.0 x 10 ３ Cells / mL or more 3.0 x 10 ９ Cells / mL or less, 1.0 x 10 ３ Cells / mL or more 1.0 x 10 ９ Cells / mL or less, 1.0 x 10 ４ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 ４ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 1.0 x 10 ４ Cells / mL or more 3.0 x 10 ９Cells / mL or less, 1.0 x 10 ４ Cells / mL or more 1.0 x 10 ９ Cells / mL or less, 1.0 x 10 ５ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 ５ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 1.0 x 10 ５ Cells / mL or more 3.0 x 10 ９ Cells / mL or less, 1.0 x 10 ５ Cells / mL or more 1.0 x 10 ９ Cells / mL or less, 1.0 x 10 ６ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 ６ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 1.0 x 10 ６ Cells / mL or more 3.0 x 10 ９ Cells / mL or less, 1.0 x 10 ７ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 1.0 x 10 ７ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 1.0 x 10 ７ Cells / mL or more 1.0 x 10 ９ Cells / mL or less, 4.0 x 10 ７ Cells / mL or more 1.0 x 10 １１ Cells / mL or less, 4.0 x 10 ７ Cells / mL or more 1.0 x 10 １０ Cells / mL or less, 4.0 x 10 ７ Cells / mL or more 1.0 x 10 ９ In this case, the daily intake of the liquid immunostimulating composition may be 10 mL to 1,000 mL, 30 mL to 800 mL, 50 mL to 500 mL, or 100 mL to 250 mL.

[0065] In the immunostimulating composition of the present embodiment, the number of bacteria having immunostimulating ability in the immunostimulating composition according to one embodiment per unit package is 1.0 × 10 ７ pcs or more, 1.0×10 ８ pcs or more, 3.0×10 ８pcs or more, 5.0×10 ８ or more or 1.0 x 10 ９ pcs or more, 5.0×10 ９ pcs or more, 1.0×10 １０ pcs or more, 5.0×10 １０ pcs or more, 1.0×10 １１ may be 1.0 × 10 or more, １４ Less than or equal to 1.0×10 １３ or less, or 1.0 x 10 １２ These upper and lower limits can be combined in any desired manner. For example, in the immunostimulating composition of this embodiment, the number of bacteria according to this embodiment per unit package may be 1.0 × 10 or less. ７ pcs or more 1.0×10 １４ Less than or equal to 1.0×10 ７ pcs or more 1.0×10 １３ Less than or equal to 1.0×10 ７ pcs or more 1.0×10 １２ Less than or equal to 1.0×10 ８ pcs or more 1.0×10 １４ Less than or equal to 1.0×10 ８ pcs or more 1.0×10 １３ Less than or equal to 1.0×10 ８ pcs or more 1.0×10 １２ Less than or equal to 3.0×10 ８ pcs or more 1.0×10 １４ Less than or equal to 3.0×10 ８ pcs or more 1.0×10 １３ Less than or equal to 3.0×10 ８ pcs or more 1.0×10 １２ Less than or equal to 5.0×10 ８ pcs or more 1.0×10 １４ Less than or equal to 5.0×10 ８ pcs or more 1.0×10 １３ Less than or equal to 5.0×10 ８ pcs or more 1.0×10 １２ Less than or equal to 1.0×10 ９ pcs or more 1.0×10 １４ Less than or equal to 1.0×10 ９ pcs or more 1.0×10 １３ Less than or equal to 1.0×10 ９ pcs or more 1.0×10 １２ Less than or equal to 5.0×10 ９pcs or more 1.0×10 １４ Less than or equal to 5.0×10 ９ pcs or more 1.0×10 １３ Less than or equal to 5.0×10 ９ pcs or more 1.0×10 １２ Less than or equal to 1.0×10 １０ pcs or more 1.0×10 １４ Less than or equal to 1.0×10 １０ pcs or more 1.0×10 １３ Less than or equal to 1.0×10 １０ pcs or more 1.0×10 １２ Less than or equal to 5.0×10 １０ pcs or more 1.0×10 １４ Less than or equal to 5.0×10 １０ 5.0 x 10 pieces or more １３ Less than or equal to 5.0×10 １０ pcs or more 1.0×10 １２ Less than or equal to 1.0×10 １１ pcs or more 1.0×10 １４ Less than or equal to 1.0×10 １１ pcs or more 1.0×10 １３ Less than or equal to 1.0×10 １１ pcs or more 1.0×10 １２ It may be less than one.

[0066] As used herein, "plant extract" refers to an extract obtained using any part of a plant and / or a processed product thereof as a raw material, and may be an extract obtained by squeezing a plant, an extract obtained by infusing a plant in a solvent, etc. "Plant extract having an effect of enhancing the immunostimulatory activity of immunostimulatory bacteria" refers to a plant extract that has the effect of enhancing the immunostimulatory activity of immunostimulatory bacteria, and may be a plant extract that, when co-cultured with the plant extract, the immunostimulatory bacteria, and immune cells, has a higher level of immunostimulatory activity than when only the immunostimulatory bacteria and immune cells are co-cultured.

[0067] The plant is not particularly limited, but may be, for example, a fruit, a root vegetable, a grain, a legume, or an herbaceous plant.

[0068] The plant used for extraction may be, for example, the whole plant, or the fruit, peel, rhizome, bulb, leaf, flower, or seed of the plant. The plant may be subjected to treatments such as roasting, aging, drying, freezing, processing, crushing, or sorting.

[0069] Examples of solvents used for extraction include aqueous media and organic solvents. "Aqueous media" refers to liquids containing water as an essential component, and examples include water, physiological saline such as phosphate-buffered saline (PBS), and microbial culture media. Examples of organic solvents include methanol and ethanol.

[0070] Whether or not the plant extract according to this embodiment has an effect of enhancing the immunostimulatory ability of bacteria is not particularly limited, and if the function of immune cells in the presence of bacteria having immunostimulatory activity is enhanced in the presence of the plant extract compared to the absence of the plant extract, it is determined that the plant extract has an effect of enhancing the immunostimulatory ability of the bacteria. As a more detailed example, whether or not the plant extract according to this embodiment has an effect of enhancing the immunostimulatory ability of bacteria having immunostimulatory activity can be confirmed by an increase in the IFN-α concentration measured by the following steps (i') to (iv') in the presence of the plant extract compared to the absence of the plant extract. (i') Red blood cell-removed mouse-derived bone marrow cells were diluted to 1 x 10 in RPMI medium prepared so that each component described below in (Culture Medium Composition) was added to the final concentration described below in (Culture Medium Composition). ６ <Culture medium composition> 10% by volume FBS 100 U / mL penicillin / streptomycin 1 mM sodium pyruvate 2.5 mM HEPES 1% by mass non-essential amino acids for MEM (NEAA) 50 μM β-mercaptoethanol 100 ng / mL Flt-3L (ii') 1 mL of the prepared cell suspension is seeded on a plate, and the plate is then heated in a CO ２ Incubated at 37°C and 5% CO ２ (iii') 2 x 10 bone marrow cells containing the induced pDCs are cultured at 4°C for 1 week to induce pDCs. ５The bacteria are suspended at a concentration of 1 mg / mL and 200 μL of each suspension is seeded onto a 96-well plate, to which 2 μL of a bacterial suspension adjusted to a concentration of 1 mg / mL with PBS is added, followed by the addition of a plant extract solution adjusted to a predetermined concentration. (iv') After 24 hours, the culture supernatant is recovered and the IFN-α concentration is measured by ELISA using an IFN-α measurement kit.

[0071] Examples of plant extracts include extracts of plants belonging to the Bromeliaceae family (e.g., pineapple extract, etc.), extracts of plants belonging to the Hypericaceae family (e.g., mangosteen extract and St. John's wort extract, etc.), extracts of plants belonging to the Brassicaceae family (e.g., maca extract, kale extract, etc.), extracts of plants belonging to the Poaceae family (e.g., fermented rice germ extract, etc.), extracts of plants belonging to the Fabaceae family (e.g., fermented soybean extract and carob extract, etc.), and extracts of plants belonging to the Asteraceae family. extracts (e.g., chrysanthemum extract, etc.), extracts of plants belonging to the Zingiberaceae family (e.g., ginger extract, etc.), extracts of plants belonging to the Cannabaceae family (e.g., hop extract, etc.), extracts of plants belonging to the Araliaceae family (e.g., red ginseng extract, etc.), extracts of plants belonging to the Garcinia family (Clusiaceae) (e.g., Garcinia extract, etc.), extracts of plants belonging to the Lamiaceae family (e.g., Rahma extract, etc.), and extracts of plants belonging to the Lauraceae family (e.g., bay laurel extract, etc.).

[0072] As the plant extract having an immunopotentiating effect, preferably, a plant extract belonging to the Bromeliaceae family (e.g., pineapple extract, etc.), a plant extract belonging to the Hypericaceae family (e.g., mangosteen extract, St. John's wort extract, etc.), a plant extract belonging to the Brassicaceae family (e.g., maca extract, etc.), a plant extract belonging to the Poaceae family (e.g., fermented rice germ extract, etc.), a plant extract belonging to the Fabaceae family (e.g., fermented soybean extract, etc.), extracts of plants belonging to the Asteraceae family (for example, chrysanthemum extract, etc.), extracts of plants belonging to the Zingiberaceae family (for example, ginger extract, etc.), extracts of plants belonging to the Cannabaceae family (for example, hop extract, etc.), extracts of plants belonging to the Araliaceae family (for example, red ginseng extract, etc.), extracts of plants belonging to the Garciniaceae family (Clusiaceae), extracts of plants belonging to the Lamiaceae family (Lamiaceae), and extracts of plants belonging to the Lauraceae family (Lauraceae).

[0073] Examples of the plant extract in the immunostimulating composition according to one embodiment include plant extracts belonging to the genus Ananas (e.g., pineapple extract, etc.), plant extracts belonging to the genus Garcinia (e.g., mangosteen extract, etc.), plant extracts belonging to the genus Lepidium (e.g., maca extract, etc.), plant extracts belonging to the genus Oryza (e.g., fermented rice germ extract, etc.), plant extracts belonging to the genus Glycine (e.g., fermented soybean extract, etc.), plant extracts belonging to the genus Hypericum (e.g., St. John's wort extract, etc.), plant extracts belonging to the genus Chrysanthemum (e.g., chrysanthemum extract, etc.), and plant extracts belonging to the genus Celastrus (e.g., celastrus extract, etc.). The extract may be an extract of a plant belonging to the genus Ceratonia (e.g., carob extract, etc.), an extract of a plant belonging to the genus Zingiber (e.g., ginger extract, etc.), an extract of a plant belonging to the genus Humulus (e.g., hop extract, etc.), an extract of a plant belonging to the genus Panax (e.g., red ginseng extract, etc.), an extract of a plant belonging to the genus Brassica (e.g., kale extract, etc.), an extract of a plant belonging to Garcinia (e.g., Garcinia extract, etc.), an extract of a plant belonging to the genus Melissa (e.g., Rahma extract, etc.), or an extract of a plant belonging to the genus Laurus (e.g., bay laurel extract, etc.).

[0074] As the plant extract having an immunopotentiating effect, preferably, a plant extract belonging to the genus Ananas (e.g., pineapple extract, etc.), a plant extract belonging to the genus Garcinia (e.g., mangosteen extract, etc.), a plant extract belonging to the genus Lepidium (e.g., maca extract, etc.), a plant extract belonging to the genus Oryza (e.g., fermented rice germ extract, etc.), a plant extract belonging to the genus Glycine (e.g., fermented soybean extract, etc.), a plant extract belonging to the genus Hypericum (e.g., St. John's wort extract, etc.), a plant extract belonging to the genus Chrysanthem ... extracts of plants belonging to the genus Brassica (e.g., chrysanthemum extract, etc.), extracts of plants belonging to the genus Ceratonia (e.g., carob extract, etc.), extracts of plants belonging to the genus Zingiber (e.g., ginger extract, etc.), extracts of plants belonging to the genus Humulus (e.g., hop extract, etc.), extracts of plants belonging to the genus Panax (e.g., red ginseng extract, etc.), extracts of plants belonging to the genus Brassica, extracts of plants belonging to Garcinia, extracts of plants belonging to the genus Melissa, and extracts of plants belonging to the genus Laurus.

[0075] Examples of the plant extract include plant extracts used as skin barrier function improving agents (e.g., pineapple extract, mangosteen extract, maca extract, etc.), plant extracts used as fatigue recovery agents (e.g., pineapple extract, maca extract, kale extract, etc.), plant extracts used as anti-aging agents (e.g., pineapple extract, maca extract, chrysanthemum extract, kale extract, etc.), plant extracts used as menopausal disorder improving agents (e.g., maca extract and hop extract, etc.), plant extracts used as arteriosclerosis prevention agents (e.g., maca extract and kale extract, etc.), plant extracts used as anti-inflammatory agents (e.g., ginger extract, Garcinia extract, Bay extract, etc.), plant extracts used as immunostimulants (e.g., ginger extract and kale extract, etc.), plant extracts used as antibacterial agents (e.g., ginger extract and hop extract, etc.), plant extracts used as body fat reducing agents (e.g., hop extract and Garcinia extract, etc.), plant extracts used as brain function improving agents (e.g., maca extract and hop extract, etc.), and the like.

[0076] Preferred examples of plant extracts having an immunostimulatory and enhancing effect include plant extracts used as skin barrier function improving agents (e.g., pineapple extract, mangosteen extract, maca extract, etc.), plant extracts used as fatigue recovery agents (e.g., pineapple extract, maca extract, etc.), plant extracts used as anti-aging agents (e.g., pineapple extract, maca extract, chrysanthemum extract, etc.), plant extracts used as menopausal disorder improving agents (e.g., maca extract, hop extract, etc.), plant extracts used as arteriosclerosis prevention agents (e.g., maca extract, etc.), plant extracts used as anti-inflammatory agents (e.g., ginger extract, etc.), plant extracts used as immunostimulants (e.g., ginger extract, etc.), plant extracts used as antibacterial agents (e.g., ginger extract, hop extract, etc.), plant extracts used as body fat reducing agents (e.g., hop extract, etc.), and plant extracts used as brain function improving agents (e.g., maca extract, hop extract, etc.).

[0077] Examples of the plant extract include plant extracts used as bittering agents (e.g., hop extracts, etc.), plant extracts used as antioxidants (e.g., mangosteen extract, ginger extract, kale extract, and Garcinia extract, etc.), plant extracts used as sweeteners (e.g., carob extract and Garcinia extract, etc.), and plant extracts used as anti-glycation agents (e.g., mangosteen extract and herb extract, etc.).

[0078] The plant extract having an immunopotentiating effect may preferably be a plant extract used as a bittering agent (e.g., hop extract, etc.), a plant extract used as an antioxidant (e.g., mangosteen extract, ginger extract, etc.), a plant extract used as a sweetener (e.g., carob extract, etc.), a plant extract used as an anti-glycation agent (e.g., mangosteen extract, herb extract, etc.), etc.

[0079] The plant extract in the immunostimulating composition according to one embodiment may be pineapple extract, mangosteen extract, maca extract, fermented extract of rice germ and soybean, St. John's wort extract, chrysanthemum extract, herb extract, carob extract, ginger extract, hop extract, red ginseng extract, kale extract, Garcinia extract, Rahma extract, bay extract, etc., and is preferably one or more plant extracts selected from the group consisting of pineapple extract, mangosteen extract, maca extract, fermented extract of rice germ and soybean, St. John's wort extract, herb extract, chrysanthemum extract, carob extract, ginger extract, hop extract, and red ginseng extract.

[0080] The pineapple extract is preferably a pineapple extract, which may be in a liquid form or a solid form such as a powder, and may be a fat-soluble component extracted by heating and refluxing pineapple in an ethanol solution of 80% by volume or more and 100% by volume or less, such as "Brightening Pine (registered trademark)" manufactured by Maruzen Pharmaceuticals.

[0081] The mangosteen extract is preferably a mangosteen peel extract, which may be in a liquid form or a solid form such as powder, and the mangosteen peel extract may be obtained by boiling mangosteen peel, drying it at 70°C for 8 hours, extracting it with hot water, filtering it, concentrating it, and powdering it, for example, "Mangosteen Aqua" manufactured by Nippon Shinyaku Co., Ltd.

[0082] The maca extract may be in liquid form or in solid form such as powder. The maca extract may be obtained by extracting maca with at least one of water and an aqueous ethanol solution, passing the extract through a high-porous synthetic polymer adsorption resin, and then adsorbing the extract onto this high-porous synthetic polymer adsorption resin. For example, MACAXS (registered trademark) manufactured by TOWA Corporation may be used.

[0083] The fermented extract of rice germ and soybeans may be in a liquid form or a solid form such as powder. The extract may be prepared by fermenting rice germ and soybeans with lactic acid bacteria and then extracting the rice germ and soybeans with at least one of water, ethanol, an aqueous ethanol solution, acetone, and hexane, without any particular limitation. For example, "Senspure (registered trademark)" manufactured by Toyo Hakko Co., Ltd. may be used.

[0084] St. John's wort (Hypericum perforatum) extract can be used in liquid or solid form such as powder, and the St. John's wort extract can be obtained by extracting St. John's wort with water, ethanol, or a mixture of water and ethanol, and for example, ``St. John's Wort'' manufactured by Fancl can be used.

[0085] The chrysanthemum extract is preferably a chrysanthemum flower extract, which may be in a liquid form or a solid form such as a powder, and may be one obtained by subjecting chrysanthemum flowers to either polar solvent extraction or supercritical extraction, and / or both, such as "Chrysanthemum Flower Extract P" manufactured by Oryza Oil & Fat Chemical Co., Ltd.

[0086] The herbal extract can be in a liquid form or a solid form such as a powder. The herbal extract is preferably a "mixed herbal extract" which is a mixture of extracts of multiple herbs. The "mixed herbal extract" can be an extract extracted with hot water from four types of herbs (chamomile, Houttuynia cordata, hawthorn, and grape leaf), and for example, "AG Herb MIX (registered trademark)" manufactured by Arkray, Inc. can be used.

[0087] The carob extract is preferably a carob pod extract. The carob extract may be in a liquid form or a solid form such as powder, and may be obtained by roasting and grinding cultivated carob pods. For example, "Organic Carob Powder" manufactured by Sakurai Foods may be used.

[0088] The ginger extract may be in a liquid form or a solid form such as a powder. Ginger extract obtained by extracting the rhizome of ginger (Zingiber officinale ROSC.) of the Zingiberaceae family with ethanol, acetone, or hexane at room temperature may be used. For example, "Ginger Extract Powder S" manufactured by Ikeda Tohka Kogyo Co., Ltd. may be used.

[0089] Hop extracts can be used in liquid or solid forms such as powder. Hop extracts can be extracted using solvents including water or various organic solvents (e.g., lower alcohols having 1 to 4 carbon atoms, such as methanol, ethanol, propanol, and butanol; lower alkyl esters such as ethyl acetate; glycols such as ethylene glycol, butylene glycol, propylene glycol, and glycerin; polar solvents such as acetone and acetic acid; hydrocarbons such as benzene and hexane; and nonpolar solvents such as ethers such as ethyl ether and petroleum ether), or extracts obtained by supercritical extraction using supercritical carbon dioxide or the like. From the viewpoint of odor removal, the extraction temperature using the solvent is preferably 60°C or less, and, taking extraction efficiency into consideration, a temperature of 50 to 60°C is more preferable. Examples of hop extracts that can be used include "Aged Hop Extract" manufactured by Kirin Holdings.

[0090] The red ginseng extract may be in a liquid form or a solid form such as powder, and the matured red ginseng extract may be obtained by subjecting the root of medicinal ginseng or an extract of the root to a heat treatment or fermentation treatment, for example, a product manufactured by Kim's Korean Ginseng.

[0091] The kale extract may be in a liquid form or a solid form such as powder. The kale extract may be prepared by washing kale, shearing or blanching the green leaves as needed, adding water as needed, and then squeezing the juice using a known method. For example, "Koi Aona (registered trademark)" manufactured by Nippon may be used.

[0092] As used herein, "Garcinia extract" refers to an extract of Garcinia cambogia (Garcinia gummi-gutta). The Garcinia extract is preferably an extract of Garcinia rind. The Garcinia extract may be in a liquid form or a solid form such as a powder. The Garcinia extract may be one containing hydroxycitric acids obtained by extracting Garcinia rind, which contains hydroxycitric acids, with water or alcohol. For example, "Garcinia Extract" manufactured by Nippon Shinyaku Co., Ltd. and "Garcinia Powder S" manufactured by Nippon Shinyaku Co., Ltd. may be used.

[0093] The Rahma extract is preferably a Rahma leaf extract, which may be in liquid or solid form such as powder, and may be obtained by extracting dried Rahma leaves with hydroethanol, followed by purification, concentration, and drying. For example, "VENETRON (registered trademark)" manufactured by Tokiwa Phytochemical Research Institute may be used.

[0094] The bay extract is preferably a bay leaf extract. The bay extract can be in liquid form or in solid form such as powder, and the bay leaf extract can be obtained by extracting and purifying dried bay leaves with, but not limited to, water, ethanol, an aqueous ethanol solution, acetone, or hexane, and for example, "LORESH (registered trademark)" manufactured by Tokiwa Phytochemical Research Institute can be used.

[0095] The effect of the plant extract according to this embodiment on enhancing the immunostimulatory ability of bacteria is such that the IFN-α concentration measured by the above steps (i') to (iv') in the presence of the plant extract is 1.01 times or more, 1.02 times or more, 1.03 times or more, 1.05 times or more, 1.08 times or more, 1.10 times or more, 1.13 times or more, 1.15 times or more, 1.18 times or more, 1.20 times or more, 1.23 times or more, as the lower limit, compared to the absence of the plant extract. Preferably, the ratio is 1.25 or more, 1.28 or more, 1.30 or more, 1.40 or more, 1.50 or more, 1.60 or more, 1.70 or more, 1.80 or more, 1.90 or more, or 2.0 or more, and the upper limit is preferably 4.0 or less, 3.8 or less, 3.5 or less, 3.3 or less, 3.0 or less, 2.8 or less, 2.5 or less, 2.3 or less, 2.0 or less, 1.80 or less, or 1.50 or less. These upper and lower limits can be combined in any desired manner. The effect of the plant extract according to this embodiment on enhancing the immunostimulatory ability of bacteria is such that, in the presence of the plant extract, the IFN-α concentration measured by the above steps (i') to (iv') is, for example, 1.01 to 4.0 times, 1.01 to 4.0 times, 1.01 to 3.8 times, 1.01 to 3.5 times, 1.01 to 3.3 times, 1.01 to 3.0 times, 1.01 to 2.8 times, 1.01 to 2.5 times, 1.01 to 2.3 times, 1.01 to 2.0 times, 1.01 to 1.8 times, 1.01 to 1.5 times, 1.02 to 4.0 times, 1.02 to 5 times 3.8 times or less, 1.02 times or more and 3.5 times or less, 1.02 times or more and 3.3 times or less, 1.02 times or more and 3.0 times or less, 1.02 times or more and 2.8 times or less, 1.02 times or more 2.5 times or less, 1.02 times or more and 2.3 times or less, 1.02 times or more and 2.0 times or less, 1.02 times or more and 1.8 times or less, 1.02 times or more and 1.5 times or less, 1.03 times or more and 4. 0 times or less, 1.03 times or more and 3.8 times or less, 1.03 times or more and 3.5 times or less, 1.03 times or more and 3.3 times or less, 1.03 times or more and 3.0 times or less, 1.03 times or more and 2.8 times or more Lower, 1.03 times or more and 2.5 times or less, 1.03 times or more and 2.3 times or less, 1.03 times or more and 2.0 times or less, 1.03 times or more and 1.8 times or less, 1.02 times or more and 1.5 times or less,1.05 times to 4.0 times, 1.05 times to 3.8 times, 1.05 times to 3.5 times, 1.05 times to 3.3 times, 1.05 times to 3.0 times, 1.05 times to 2.8 times, 1.05 times to 2.5 times, 1.05 times to 2.3 times, 1.05 times to 2.0 times, 1.05 times to 1.8 times, 1.05 times to 1.5 times, 1.08 times to 4.0 times, 1.08 times to 3.8 times, 1.08 times to 3.5 times, 1.08 times to 3.3 times, 1.08 times to 3.0 times, 1.08 times to 2.8 times, 1.08 times to 2.5 times Below, 1.08 times to 2.3 times, 1.08 times to 2.0 times, 1.08 times to 1.8 times, 1.08 times to 1.5 times, 1.10 times to 4.0 times, 1.10 times to 3.8 times, 1.10 times to 3.5 times, 1.10 times to 3.3 times, 1.10 times to 3.0 times, 1.10 times to 2.8 times, 1.10 times to 2.5 times, 1.10 times to 2.3 times, 1.10 times to 2.0 times, 1.10 times to 1.8 times, 1.10 times to 1.5 times, 1.13 times to 4.0 times, 1.13 times to 3.8 times, 1.13 times and above. Below 3.5 times, 1.13 times to 3.3 times, 1.13 times to 3.0 times, 1.13 times to 2.8 times, 1.13 times to 2.5 times, 1.13 times to 2.3 times, 1.13 times to 2.0 times, 1.13 times to 1.8 times, 1.13 times to 1.5 times, 1.15 times to 4.0 times, 1.15 times to 3.8 times, 1.15 times to 3.5 times, 1.15 times to 3.3 times, 1.15 times to 3.0 times, 1.15 times to 2.8 times, 1.15 times to 2.5 times, 1.15 times to 2.3 times, 1.15 times to 2.0 times, 1.15 times to 2.0 times. The following are considered high-risk bets: 1.15 to 1.5 times, 1.18 to 4.0 times, 1.18 to 3.8 times, 1.18 to 3.5 times, 1.18 to 3.3 times, 1.18 to 3.0 times, 1.18 to 2.8 times, 1.18 to 2.5 times, 1.18 to 2.3 times, 1.18 to 2.0 times, 1.18 to 1.8 times, 1.18 to 1.5 times, 1.20 to 4.0 times, 1.20 to 3.8 times, 1.20 to 3.5 times, 1.20 to 3.3 times, and 1.20 to 3.0 times.1.20 times or more but less than 2.8 times, 1.20 times or more but less than 2.5 times, 1.20 times or more but less than 2.3 times, 1.20 times or more but less than 2.0 times, 1.20 times or more but less than 1.8 times, 1.20 times or more but less than 1.5 times, 1.23 times or more but less than 4.0 times, 1.23 times or more but less than 3.8 times, 1.23 times or more but less than 3.5 times, 1.23 times or more but less than 3.3 times, 1.23 times or more but less than 3.0 times, 1.23 times or more but less than 2.8 times, 1.23 times or more but less than 2.5 times, 1.23 times or more but less than 2.3 times, 1.23 times or more but less than 2.0 times, 1.23 times or more but less than 1.8 times, 1.23 times or more but less than 1.5 times, 1.25 times or more but less than 4.0 times. Below 1.25 times, 1.25 times to 3.8 times, 1.25 times to 3.5 times, 1.25 times to 3.3 times, 1.25 times to 3.0 times, 1.25 times to 2.8 times, 1.25 times to 2.5 times, 1.25 times to 2.3 times, 1.25 times to 2.0 times, 1.25 times to 1.8 times, 1.25 times to 1.5 times, 1.28 times to 4.0 times, 1.28 times to 3.8 times, 1.28 times to 3.5 times, 1.28 times to 3.3 times, 1.28 times to 3.0 times, 1.28 times to 2.8 times, 1.28 times to 2.5 times, 1.28 times and above. Below 2.3 times, 1.28 times to below 2.0 times, 1.28 times to below 1.8 times, 1.28 times to below 1.5 times, 1.30 times to below 4.0 times, 1.30 times to below 3.8 times, 1.30 times to below 3.5 times, 1.30 times to below 3.3 times, 1.30 times to below 3.0 times, 1.30 times to below 2.8 times, 1.30 times to below 2.5 times, 1.30 times to below 2.3 times, 1.30 times to below 2.0 times, 1.30 times to below 1.8 times, 1.30 times to below 1.5 times, 1.40 times to below 4.0 times, 1.40 times to below 3.8 times, 1.40 times to below 3.5 times, 1.4 0 to 3.3 times, 1.40 to 3.0 times, 1.40 to 2.8 times, 1.40 to 2.5 times, 1.40 to 2.3 times, 1.40 to 2.0 times, 1.40 to 1.8 times, 1.40 to 1.5 times, 1.50 to 4.0 times, 1.50 to 3.8 times, 1.50 to 3.5 times, 1.50 to 3.3 times, 1.50 to 3.0 times, 1.50 to 2.8 times, 1.50 to 2.5 times, 1.50 to 2.3 times, 1.50 to 2.0 times, 1.50 to 1.8 times.1.60 times to 4.0 times, 1.60 times to 3.8 times, 1.60 times to 3.5 times, 1.60 times to 3.3 times, 1.60 times to 3.0 times, 1.60 times to 2.8 times 1.60 times or more and 2.5 times or less, 1.60 times or more and 2.3 times or less, 1.60 times or more and 2.0 times or less, 1.60 times or more and 1.8 times or less, 1.70 times or more and 4.0 times or less, 1.70 times or more and 3. 8 times or less, 1.70 times or more and 3.5 times or less, 1.70 times or more and 3.3 times or less, 1.70 times or more and 3.0 times or less, 1.70 times or more and 2.8 times or less, 1.70 times or more and 2.5 times or less, 1.70 times or more 2.3 times or less, 1.70 times or more and 2.0 times or less, 1.70 times or more and 1.8 times or less, 1.80 times or more and 4.0 times or less, 1.80 times or more and 3.8 times or less, 1.80 times or more and 3.5 times or less, 1.80 times 3.3 times or more, 1.80 times or more and 3.0 times or less, 1.80 times or more and 2.8 times or less, 1.80 times or more and 2.5 times or less, 1.80 times or more and 2.3 times or less, 1.80 times or more and 2.0 times or less, 1. 90 times or more and 4.0 times or less, 1.90 times or more and 3.8 times or less, 1.90 times or more and 3.5 times or less, 1.90 times or more and 3.3 times or less, 1.90 times or more and 3.0 times or less, 1.90 times or more and 2.8 times or less, Preferably, the ratio is 1.90 to 2.5 times, 1.90 to 2.3 times, 1.90 to 2.0 times, 2.00 to 4.0 times, 2.00 to 3.8 times, 2.00 to 3.5 times, 2.00 to 3.3 times, 2.00 to 3.0 times, 2.00 to 2.8 times, 2.00 to 2.5 times, or 2.00 to 2.3 times.

[0096] The content of the plant extract in the immunostimulating composition according to one embodiment can be, for example, a lower limit of 0.000001% by mass or more, 0.00001% by mass or more, 0.001% by mass or more, 0.01% by mass or more, 0.02% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.5% by mass or more, 1% by mass or more, or 2% by mass or more, and an upper limit of 90.0% by mass or more, based on the total amount of the immunostimulating composition. The upper limit and lower limit may be 85.0% by mass or less, 80.0% by mass or less, 75.0% by mass or less, 50.0% by mass or less, 25.0% by mass or less, 10.0% by mass or less, 5.0% by mass or less, 1.0% by mass or less, 0.80% by mass or less, 0.50% by mass or less, 0.30% by mass or less, 0.25% by mass or less, 0.20% by mass or less, 0.10% by mass or less, 0.05% by mass or less, 0.02% by mass or less, or 0.01% by mass or less. These upper and lower limits may be combined in any desired manner. The content of the plant extract in the immunostimulating composition according to one embodiment is, for example, 0.000001% by mass to 90.0% by mass, 0.000001% by mass to 85.0% by mass, 0.000001% by mass to 80.0% by mass, 0.000001% by mass to 75.0% by mass, 0.000001% by mass to 50.0% by mass, 0.000001% by mass to 25.0% by mass, 0.000001% by mass to 10.0% by mass, 0.000001% by mass to 5.0% by mass, 0.000001% by mass to 1.0% by mass, 0.000001% by mass to 0.80% by mass, or 0.000001% by mass to 0.50% by mass, relative to the total amount of the immunostimulating composition. mass% or less, 0.000001 mass% or more and 0.30 mass% or less, 0.000001 mass% or more and 0.25 mass% or less, 0.000001 mass% or more and 0.20 mass% or less, 0.000001 mass% or more and 0.10 mass% or less, 0.000001 mass% or more and 0.05 mass% or less, 0.000001 mass% or more and 0.02 mass% or less, 0.0000 01 mass% or more and 0.01 mass% or less, 0.00001 mass% or more and 90.0 mass% or less, 0.00001 mass% or more and 85.0 mass% or less, 0.00001 mass% or more8 0.0 mass% or less, 0.00001 mass% or more and 75.0 mass% or less, 0.00001 mass% or more and 50.0 mass% or less, 0.00001 mass% or more and 25.0 mass% or less,0.00001% to 10.0% of mass, 0.00001% to 5.0% of mass, 0.00001% to 1.0% of mass, 0.00001% to 0.80% of mass, 0.00001% to 0.50% of mass, 0.00001% to 0.30% of mass, 0.00001% to 0.25% of mass, 0.00001% to 0.20% of mass, 0.00001% to 0.10% of mass, 0.00001% to 0.05% of mass, 0.00001% to 0.02% of mass, 0.00001% of mass... 0.01% to 0.01% of mass, 0.001% to 90.0% of mass, 0.001% to 85.0% of mass, 0.001% to 80.0% of mass, 0.001% to 75.0% of mass, 0.001% to 50.0% of mass, 0.001% to 25.0% of mass, 0.001% to 10.0% of mass, 0.001% to 5.0% of mass, 0.001% to 1.0% of mass, 0.001% to 0.80% of mass, 0.001% to 0.50% of mass, 0.001% to 0.30% of mass. 0.001% to 0.25% of mass, 0.001% to 0.20% of mass, 0.001% to 0.10% of mass, 0.001% to 0.05% of mass, 0.001% to 0.02% of mass, 0.001% to 0.01% of mass, 0.01% to 90.0% of mass, 0.01% to 85.0% of mass, 0.01% to 80.0% of mass, 0.01% to 75.0% of mass, 0.01% to 50.0% of mass, 0.01% to 25.0% of mass, 0.01% to 10.0% of mass. Less than 1% of mass, 0.01% to 5.0% of mass, 0.01% to 1.0% of mass, 0.01% to 0.80% of mass, 0.01% to 0.50% of mass, 0.01% to 0.30% of mass, 0.01% to 0.25% of mass, 0.01% to 0.20% of mass, 0.01% to 0.10% of mass, 0.01% to 0.05% of mass, 0.01% to 0.02% of mass, 0.02% to 90.0% of mass, 0.02% to 85.0% of mass, 0.02% to 80.0% of mass.0.02% to 75.0% by mass, 0.02% to 50.0% by mass, 0.02% to 25.0% by mass, 0.02% to 10.0% by mass, 0.02% to 5.0% by mass, 0.02% to 1.0% by mass, 0.02% to 0.80% by mass, 0.02% to 0.50% by mass, 0.02% to 0.30% by mass, 0.02% to 0.25% by mass, 0.02% to 0.20% by mass, 0.02% to 0.10% by mass, 0.02% to 0.05% by mass, 0.05% by mass. Quality percentage above 90.0% and below 90.0%; 0.05% and above 85.0% and below 80.0%; 0.05% and above 75.0% and below 75.0%; 0.05% and above 50.0% and below 50.0%; 0.05% and above 25.0% and below 25.0%; 0.05% and above 10.0% and below 10.0%; 0.05% and above 5.0% and below 1.0%; 0.05% and above 0.80% and below 0.05% and below 0.50%; 0.05% and above 0.30% and below 0.05% and below 0.25%; 0.05% and below 10.0% of the total mass. Above 0.20% by mass, 0.05% by mass and below 0.10% by mass, 0.10% by mass and below 90.0% by mass, 0.10% by mass and below 85.0% by mass, 0.10% by mass and below 80.0% by mass, 0.10% by mass and below 75.0% by mass, 0.10% by mass and below 50.0% by mass, 0.10% by mass and below 25.0% by mass, 0.10% by mass and below 10.0% by mass, 0.10% by mass and below 5.0% by mass, 0.10% by mass and below 1.0% by mass, 0.10% by mass and below 0.80% by mass, 0.10% by mass and below 0.50% by mass, 0.10% by mass and above 0.3% by mass. Less than 0% by mass, 0.10% to 0.25% by mass, 0.10% to 0.20% by mass, 0.50% to 90.0% by mass, 0.50% to 85.0% by mass, 0.50% to 80.0% by mass, 0.50% to 75.0% by mass, 0.50% to 50.0% by mass, 0.50% to 25.0% by mass, 0.50% to 10.0% by mass, 0.50% to 5.0% by mass, 0.50% to 1.0% by mass, 0.50% to 0.80% by mass, 1% to 90.0% by mass.1% or more of quality, less than 85.0% of quality; 1% or more of quality, less than 80.0% of quality; 1% or more of quality, less than 75.0% of quality; 1% or more of quality, less than 50.0% of quality; 1% or more of quality, less than 25.0% of quality; 1% or more of quality, less than 10.0% of quality; 1% or more of quality, less than 5.0% of quality; 2% or more of quality, less than 90.0% of quality; 2% or more of quality, less than 85.0% of quality; 2% or more of quality, less than 80.0% of quality; 2% or more of quality, less than 75.0% of quality; 2% or more of quality, less than 50.0% of quality; 2% or more of quality, less than 25.0% of quality; 2% or more of quality, less than 10.0% of quality; 2% or more of quality, less than 5.0% of quality.

[0097] The ratio of the content (% by mass) of bacteria having immunostimulatory ability to the content (% by mass) of plant extract in the immunostimulatory composition according to one embodiment ((content of bacteria having immunostimulatory ability) / (content of plant extract)) can have a lower limit of, for example, 0.001 or more, 0.003 or more, 0.005 or more, 0.01 or more, 0.015 or more, 0.02 or more, 0.025 or more, or 0.03 or more, and an upper limit of 50,000 or less, 10,000 or less, 1,000 or less, 500 or less, 100 or less, 50 or less, 10 or less, 5 or less, 1.0 or less, 0.75 or less, 0.5 or less, or 0.3 or less. These upper and lower limits can be combined in any desired manner. The ratio of the content (mass%) of bacteria having immunostimulatory ability to the content (mass%) of plant extract in the immunostimulatory composition according to one embodiment ((content of bacteria having immunostimulatory ability) / (content of plant extract)) is, for example, 0.001 or more and 50,000 or less, 0.001 or more and 10,000 or less, 0.001 or more and 1,000 or less, 0.001 or more and 500 or less, 0.001 or more and 100 or less, 0.00 1 to 50, 0.001 to 10, 0.001 to 5, 0.001 to 1.0, 0.001 to 0.75, 0.001 to 0.5, 0.001 to 0.3, 0.0 03 to 50,000, 0.003 to 10,000, 0.003 to 1,000, 0.003 to 500, 0.003 to 100, 0.003 to 50, 0.003 to 1 0 or less, 0.003 or more and 5 or less, 0.003 or more and 1.0 or less, 0.003 or more and 0.75 or less, 0.003 or more and 0.5 or less, 0.003 or more and 0.3 or less, 0.005 or more and 50000 or less, 0.00 5 to 10000, 0.005 to 1000, 0.005 to 500, 0.005 to 100, 0.005 to 50, 0.005 to 10, 0.005 to 5, 0 .. 005 to 1.0, 0.005 to 0.75, 0.005 to 0.5, 0.005 to 0.3, 0.01 to 50,000, 0.01 to 10,000, 0.01 to 1 000 or less, 0.01 or more and 500 or less, 0.01 or more and 100 or less, 0.01 or more and 50 or less, 0.01 or more and 10 or less, 0.01 or more and 5 or less, 0.01 or more and 1.0 or less, 0.01 or more and 0.75 or less,0.01 to 0.5, 0.01 to 0.3, 0.015 to 50000, 0.015 to 10000, 0.015 to 1000, 0.015 to 500 Lower, 0.015 to 100, 0.015 to 50, 0.015 to 10, 0.015 to 5, 0.015 to 1.0, 0.015 to 0.75, 0.0 15 to 0.5, 0.015 to 0.3, 0.02 to 50,000, 0.02 to 10,000, 0.02 to 1,000, 0.02 to 500, 0.02 100 or more, 0.02 or more and 50 or less, 0.02 or more and 10 or less, 0.02 or more and 5 or less, 0.02 or more and 1.0 or less, 0.02 or more and 0.75 or less, 0.02 or more and 0.5 or less, 0.0 2 to 0.3, 0.025 to 50,000, 0.025 to 10,000, 0.025 to 1,000, 0.025 to 500, 0.025 to 100, 0 .025 to 50, 0.025 to 10, 0.025 to 5, 0.025 to 1.0, 0.025 to 0.75, 0.025 to 0.5 It may be 0.3 or less, 0.03 or more and 50,000 or less, 0.03 or more and 10,000 or less, 0.03 or more and 1,000 or less, 0.03 or more and 500 or less, 0.03 or more and 100 or less, 0.03 or more and 50 or less, 0.03 or more and 10 or less, 0.03 or more and 5 or less, 0.03 or more and 1.0 or less, 0.03 or more and 0.75 or less, 0.03 or more and 0.5 or less, or 0.03 or more and 0.3 or less.

[0098] The immunostimulatory composition according to one embodiment may be a food (including a beverage), a drug, or a quasi-drug. The food, drug, or quasi-drug can each be produced according to a conventional method. The content of the immunostimulatory bacteria and plant extract in the food, drug, or quasi-drug is not particularly limited and can be freely set depending on the purpose.

[0099] When the immunostimulating composition according to one embodiment is a food, drug, or quasi-drug, the food, drug, or quasi-drug may contain, in addition to the immunostimulating bacteria and plant extract, ingredients typically used in foods, drugs, or quasi-drugs. The food, drug, or quasi-drug according to one embodiment may contain bases, carriers, additives, etc. typically used in foods, drugs, or quasi-drugs. Examples of additives include excipients, oils, powders, buffers, solubilizers, antioxidants, surfactants, thickeners, preservatives, pH adjusters, chelating agents, stabilizers, irritation reducers, antiseptics, pigments, colorants, fragrances, gloss-imparting agents, gelling agents, alcohols, water-soluble polymers, film-forming agents, resins, etc. The bases, carriers, and various additives described above may be used singly or in combination, as needed.

[0100] When the immunostimulating composition according to one embodiment is a food product, examples of the food product include health foods, functional foods, nutritional compositions, dietary supplements, supplements, health foods, foods for specified health uses, foods with nutrient functions, and foods with functional claims. Such food compositions can be labeled, for example, as supporting the maintenance of immune function in healthy individuals (immune care), or suppressing immune function decline for individuals concerned about a decline in immune function. The immunostimulating composition according to one embodiment can also be used as a food additive.

[0101] Examples of such foods include seasonings, processed meat products, processed agricultural products, beverages (lactic acid bacteria drinks, soft drinks, alcoholic drinks, carbonated drinks, dairy drinks, fruit juice drinks, tea, coffee, energy drinks, etc.), powdered drinks (powdered juice, powdered soup, powdered milk, etc.), concentrated drinks, confectioneries (candy (throat lozenges), cookies, biscuits, gum, gummy candies, chewable tablets, tablets, chocolate, etc.), bread, cereal, etc. In addition, in the case of foods for specified health uses, foods with nutrient functions, foods with functional claims, etc., the foods may be in the form of capsules, granules, powders, syrups, lozenges, etc.

[0102] When the immunostimulating composition according to one embodiment is a pharmaceutical or quasi-drug, the pharmaceutical or quasi-drug may be in the form of, for example, a liquid, suspension, capsule, granule, pill, powder, tablet, syrup, lozenge, etc. Possible indications for the pharmaceutical include allergies, obesity, heart failure, etc.

[0103] The immunostimulatory composition according to one embodiment is preferably administered or ingested into the body. The mode of administration or ingestion may be oral administration or ingestion or parenteral administration or ingestion, with oral administration or ingestion being preferred. The immunostimulatory composition according to one embodiment may be administered or ingested only once or multiple times, and, if sustained immunostimulation is desired, is preferably administered or ingested continuously or intermittently over a certain period of time. The food composition according to one embodiment may be ingested, for example, 1 to 5 times a day, once every 2 days, once every 3 days, once every 4 days, or once a week for one week or more, two weeks or more, one month or more, three months or more, six months or more, one year or more, three years or more, five years or more, or ten years or more.

[0104] The immunostimulatory composition according to one embodiment is an immunostimulatory composition induced by bacteria having immunostimulatory ability, and comprises a plant extract having an effect of enhancing the immunostimulatory ability of the bacteria. "Immunostimulation induced by bacteria having immunostimulatory ability" refers to immunostimulation induced depending on the ability of bacteria having immunostimulatory ability to activate immune cells, which is inherent in the bacteria.

[0105] The immunostimulating composition according to one embodiment is a composition for immunostimulation or a composition for immunostimulation with enhanced immunostimulating activity, which is prepared by adding a plant extract having an effect of enhancing the immunostimulating activity of a bacterium having immunostimulating activity to the bacterium. The immunostimulating composition or the immunostimulating composition with enhanced immunostimulating activity is preferable from the viewpoint that the addition of the plant extract enhances the immunostimulating activity of the bacterium.

[0106] [Enhancer of immunostimulatory effect induced by bacteria having immunostimulatory ability] A second embodiment of the present disclosure relates to an enhancer of immunostimulatory effect induced by bacteria having immunostimulatory ability (hereinafter also simply referred to as "enhancer"). One embodiment of the enhancer comprises bacteria having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacteria. Another embodiment of the enhancer is an enhancer of immunostimulatory effect induced by bacteria having immunostimulatory ability, and comprises a plant extract having an effect of enhancing the immunostimulatory ability of the bacteria. The enhancer according to this embodiment has an effect of enhancing the immunostimulatory effect induced by the bacteria having immunostimulatory ability. In other words, the enhancer according to this embodiment has an enhanced immunostimulatory effect compared to an immunostimulatory composition containing only bacteria having immunostimulatory ability.

[0107] In one embodiment, the enhancer has an ability to induce interferon α production in immune cells that is, for example, 1.01 times or more, 1.02 times or more, 1.03 times or more, 1.05 times or more, 1.08 times or more, 1.10 times or more, 1.13 times or more, 1.15 times or more, 1.18 times or more, 1.20 times or more, 1.23 times or more, 1.25 times or more, or 1.30 times or more, as a lower limit, compared to an immunostimulating composition containing only bacteria having immunostimulating ability. Preferably, the saturation concentration is 28-fold or more, 1.30-fold or more, 1.40-fold or more, 1.50-fold or more, 1.60-fold or more, 1.70-fold or more, 1.80-fold or more, 1.90-fold or more, or 2.0-fold or more, with the upper limit being preferably 4.0-fold or less, 3.8-fold or less, 3.5-fold or less, 3.3-fold or less, 3.0-fold or less, 2.8-fold or less, 2.5-fold or less, 2.3-fold or less, 2.0-fold or less, 1.8-fold or less, or 1.50-fold or less. These upper and lower limits can be arbitrarily combined and may be the same as the ranges described for the bacterial immunostimulatory activity enhancing effect of the plant extract according to this embodiment.

[0108] As other specific aspects of the enhancer according to this embodiment, aspects of the immunostimulating composition according to this embodiment can be appropriately applied.

[0109] [Immunostimulating method, method for enhancing immunostimulating effect induced by bacteria having immunostimulating ability] A third embodiment of the present disclosure relates to an immunostimulating method and a method for enhancing immunostimulating effect induced by bacteria having immunostimulating ability (hereinafter also simply referred to as "enhancing method"). The immunostimulating method and enhancing method include administering or ingesting to a subject bacteria having immunostimulating ability and a plant extract having an immunostimulating ability enhancing effect. The enhancing method according to this embodiment enhances immunostimulating effect compared to when only bacteria having immunostimulating ability are administered or ingested.

[0110] The subject of the above method can be a human or a non-human mammal, preferably a human. Examples of non-human mammals include a mouse, rat, guinea pig, hamster, rabbit, cat, dog, sheep, pig, cow, horse, goat, and monkey.

[0111] The subject can be a subject in need of immunostimulation, and the subject in need of immunostimulation is not particularly limited, but examples include subjects infected with a virus, subjects wishing to prevent viral infection, subjects with a cold, subjects wishing to prevent colds, and subjects aged 65 or older.

[0112] The bacteria having immunostimulatory activity and the plant extract having the effect of enhancing the immunostimulatory activity of the bacteria can be administered or ingested by a subject, for example, in the form of the composition of the first embodiment described above. Generally, pharmaceutical compositions are expressed as "administered to a subject," and food compositions are expressed as "ingested by a subject." These expressions are synonymous, meaning that the composition is taken into the body of the subject. Therefore, the two expressions are interchangeable and there is no essential difference between them.

[0113] [Method for producing a composition] A fourth embodiment of the present disclosure relates to a method for producing a composition for immunostimulation. This production method is a method for producing a composition for immunostimulation or a composition for immunostimulation with enhanced immunostimulatory activity, by adding a plant extract having an effect of enhancing the immunostimulatory activity of a bacterium having immunostimulatory activity to the bacterium. The immunostimulatory composition or the immunostimulatory composition with enhanced immunostimulatory activity is preferable from the viewpoint that the addition of the plant extract enhances the immunostimulatory activity of the bacterium.

[0114] [Use of bacteria and plant extract for producing a composition] A fifth embodiment of the present disclosure relates to the use of bacteria and a plant extract having the effect of enhancing the immunostimulatory ability of the bacteria for producing a composition. One aspect of use is the use of bacteria having immunostimulatory ability and a plant extract having the effect of enhancing the immunostimulatory ability of the bacteria for producing a composition for immunostimulation. Another aspect of use is the use of a plant extract having the effect of enhancing the immunostimulatory ability of the bacteria for producing a composition for immunostimulation that is induced by the bacteria having immunostimulatory ability. Yet another aspect of use is the use of a plant extract having the effect of enhancing the immunostimulatory ability of the bacteria for producing a composition for immunostimulation that is administered or ingested in combination with the bacteria having immunostimulatory ability. Another aspect of use is the use of bacteria having immunostimulatory ability for producing a composition for immunostimulation that is administered or ingested in combination with the bacteria having immunostimulatory ability. Specific aspects of the composition for immunostimulation, bacteria, and plant extract according to this embodiment are as described above.

[0115] [Plant Extract for Use] A sixth embodiment of the present disclosure relates to a plant extract having an effect of enhancing the immunostimulatory activity of a bacterium having immunostimulatory activity, for use in immunostimulation induced by the bacterium. Specific aspects of the bacterium and plant extract according to this embodiment are as described above.

[0116] [Bacteria or Plant Extract for Use in a Therapeutic Method] A seventh embodiment of the present disclosure relates to the use of bacteria or plant extracts for use in an immunostimulatory therapeutic method. In one aspect, the bacteria have immunostimulatory activity for use in an immunostimulatory therapeutic method, and are administered or ingested in combination with a plant extract having an effect of enhancing the immunostimulatory activity of the bacteria. In another aspect, the plant extract has an effect of enhancing the immunostimulatory activity of bacteria having immunostimulatory activity, and is administered or ingested in combination with the bacteria having immunostimulatory activity for use in an immunostimulatory therapeutic method. Specific aspects of the bacteria and plant extracts according to this embodiment are as described above. In this disclosure, use in a "therapeutic" method refers to use in treating the body of a subject. Use in a therapeutic method according to the present disclosure may be, for example, use for the purpose of or accompanying medical treatment. Use in a therapeutic method according to the present disclosure may involve, for example, a medical professional administering or ingesting a substance to a human or animal, or instructing a human or animal to administer or ingest a substance. The use in the therapeutic method according to the present disclosure may be, for example, for therapeutic or prophylactic purposes, for therapeutic purposes, or for prophylactic purposes involving ingestion of the food composition, or for therapeutic purposes. The use in the therapeutic method according to the present disclosure may be, for example, for use in unhealthy individuals.

[0117] [Use of bacteria or plant extract in non-therapeutic methods] The eighth embodiment of the present disclosure relates to the use of bacteria or plant extracts for use in non-therapeutic methods of immunostimulation. In one aspect, the use is of bacteria having immunostimulatory ability in non-therapeutic methods of immunostimulation, where the bacteria are administered or ingested in combination with a plant extract having an effect of enhancing the immunostimulatory ability of the bacteria. In another aspect, the use is of a plant extract having an effect of enhancing the immunostimulatory ability of the bacteria in non-therapeutic methods of immunostimulation, where the bacteria are administered or ingested in combination with the bacteria having immunostimulatory ability. Specific aspects of the bacteria and plant extracts according to this embodiment are as described above. In the present disclosure, use in a "non-therapeutic" method refers to use of a substance that does not fall under the category of use in a therapeutic method. Use in a non-therapeutic method according to the present disclosure may, for example, be use that is not intended for and / or does not involve medical treatment. Use in non-therapeutic methods according to the present disclosure may not involve, for example, a medical professional administering or having a substance be taken by a human or animal, and / or instructing a human or animal to administer or take a substance. Use in non-therapeutic methods according to the present disclosure may be, for example, use for preventive or health promotion purposes, and may be use for preventive or health promotion purposes involving the administration or intake of a pharmaceutical composition or quasi-drug. Use in non-therapeutic methods according to the present disclosure may be, for example, use in healthy individuals.

[0118] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to the following examples.

[0119] The plant extracts shown in Table 1 were examined for their enhancing or suppressing effects on IFN production by pDC induced by JCM5805.

[0120] <1. Preparation of Lactic Acid Bacteria Suspension> JCM5805 bacteria were inoculated into 10 mL of MRS medium (Oxoid) and cultured at 30°C for 24 hours. Then, 1 mL of the cultured lactic acid bacteria cell suspension was inoculated into 100 mL of MRS medium and cultured at 30°C for 24 hours. The lactic acid bacteria were then collected, washed three times with sterile water, heat-sterilized at 80°C for 30 minutes, and freeze-dried to obtain heat-killed bacteria. The heat-killed bacteria were then adjusted to a concentration of 1 mg / mL with PBS (Takara Bio Inc.) to prepare a lactic acid bacteria suspension.

[0121] 2. Preparation of Plant Extract Solutions The pineapple fruit extract was "Brightening Pine (registered trademark)" manufactured by Maruzen Pharmaceuticals, the mangosteen peel extract was "Mangosteen Aqua" manufactured by Nippon Shinyaku, the maca extract was "MACAXS (registered trademark)" manufactured by TOWA, the rice germ and fermented soybean extract was "Senspure (registered trademark)" manufactured by Toyo Hakko, the St. John's Wort (Hypericum perforatum) extract was "St. John's Wort" manufactured by FANCL, the chrysanthemum flower extract was "Chrysanthemum Flower Extract P" manufactured by Oryza Oil & Fat Chemical, and the herb (mixed herb of chamomile, Houttuynia cordata, hawthorn, and grape leaf) extract was "Peach Juice" manufactured by Arley. The plant extracts used were "AG Herb MIX (registered trademark)" manufactured by Sakurai Foods, "Organic Carob Powder" manufactured by Sakurai Foods for the carob extract, "Ginger Extract Powder S" manufactured by Ikeda Tohka Kogyo for the ginger extract, a product manufactured by Kirin for the matured hop extract, a product manufactured by Kimji Ginseng for the matured red ginseng extract, "Koi Aona (registered trademark)" manufactured by Nippun for the kale extract, "Garcinia Powder S" manufactured by Nippon Shinyaku for the garcinia peel extract, "Venetron (registered trademark)" manufactured by Tokiwa Plant Science Research for the rafuma leaf extract, and "Lorresh (registered trademark)" manufactured by Tokiwa Plant Science Research for the bay leaf extract. These plant extracts were adjusted to 400 μg / ml with PBS to prepare plant extract solutions.

[0122] 3. Test Procedure The test was carried out according to the following procedures (i) to (iv): (i) Bone marrow cells derived from female BALB / c mice were cultured at a concentration of 1 x 10 in RPMI medium (Sigma Corporation) prepared so that the components listed below (Culture medium composition) were added to the final concentrations listed below (Culture medium composition). ６(ii) 1 mL of the prepared cell suspension was seeded on the cells, and the cells were suspending at a concentration of 10 cells / mL to prepare a cell suspension. (Culture medium composition) 10% by volume FBS 100 U / mL penicillin / streptomycin 1 mM sodium pyruvate 2.5 mM HEPES 1% by mass non-essential amino acids for MEM (NEAA) 50 μM β-mercaptoethanol 100 ng / mL Flt-3L ２ Incubated at 37°C and 5% CO ２ (iii) The bone marrow cells containing the induced pDCs were cultured at 2 × 10 ５ The cells were suspended at a concentration of 100 cells / mL and seeded in 200 μL aliquots onto a 96-well plate. 2 μL of the JCM5805 suspension (described above in 1.) was added, and simultaneously 5 μL of the plant extract solution (described above in 2.) was added. Similarly, the plant extract solution alone was added to a 96-well plate seeded with bone marrow cells containing pDC. Similarly, the JCM5805 suspension alone was added to a 96-well plate seeded with bone marrow cells containing pDC. (iv) After 24 hours, the culture supernatant was collected, and the IFN-α concentration was measured using an IFN-α measurement kit (PBL Assay Science Ltd.).

[0123] The amount of IFN-α produced in the presence of each substance is shown in the table below relative to the amount of IFN-α produced when JCM5805 suspension was used alone, which was set at 1. The concentration of each substance in the table means the final concentration.

[0124]

[0125] When any of the substances was used alone, IFN-α production by pDC was not induced. Kale extract, Garcinia cambogia peel extract, Rafuma leaf extract, and Bay leaf extract suppressed JCM5805-induced IFN-α production by pDC, while pineapple fruit extract, mangosteen peel extract, maca extract, fermented rice germ and soybean extract, St. John's wort extract, chrysanthemum flower extract, herb extract, carob pod extract, ginger extract, hop extract, and aged red ginseng extract enhanced JCM5805-induced IFN-α production by pDC.

Claims

1. A composition for immunostimulation comprising a bacterium having immunostimulatory activity and a plant extract having an effect of enhancing the immunostimulatory activity of the bacterium.

2. The immunostimulating composition according to claim 1, wherein the bacteria having immunostimulating ability are lactic acid bacteria and / or acetic acid bacteria.

3. The immunostimulating composition according to claim 2, wherein the lactic acid bacteria are Lactococcus bacteria.

4. The immunostimulatory composition according to claim 3, wherein the Lactococcus bacterium is Lactococcus lactis subsp. lactis.

5. The immunostimulating composition according to claim 1, wherein the content of the bacteria having immunostimulating ability is 0.001% by mass or more and less than 100% by mass relative to the total amount of the immunostimulating composition.

6. The immunostimulating composition according to claim 1, wherein the content of the plant extract is 0.000001% by mass or more and 90.0% by mass or less relative to the total amount of the immunostimulating composition.

7. The immunostimulating composition according to claim 1, wherein the ratio of the content (mass%) of the bacteria having immunostimulating ability to the content (mass%) of the plant extract (content of plant extract) / (content of bacteria having immunostimulating ability) is 0.001 or more and 50,000 or less.

8. The immunostimulating composition according to any one of claims 1 to 7, wherein the immunostimulating ability is the ability to induce interferon α production in immune cells.

9. The immunostimulatory composition according to claim 8, wherein the immune cells are plasmacytoid dendritic cells.

10. The immunostimulating composition according to any one of claims 1 to 7, wherein the immunostimulating effect is an effect of enhancing the immunostimulating effect by 1.10 times or more.

11. The immunostimulating composition according to claim 8, wherein the plant extract is one or more plant extracts selected from the group consisting of pineapple extract, mangosteen extract, maca extract, fermented rice germ and soybean extract, St. John's wort extract, chrysanthemum extract, herb extract, carob extract, ginger extract, hop extract, and red ginseng extract.

12. A method for immunostimulation, comprising administering or having a subject ingest a bacterium having immunostimulatory ability and a plant extract having an effect of enhancing the immunostimulatory ability of the bacterium.

13. A method for enhancing the immunostimulatory effect induced by bacteria having immunostimulatory ability, comprising administering or having a subject ingest bacteria having immunostimulatory ability and a plant extract having the effect of enhancing the immunostimulatory ability of the bacteria.

14. The method according to claim 12 or 13, wherein the immunostimulatory activity is the ability to induce interferon α production in immune cells.

15. An agent for enhancing the immunostimulating effect induced by bacteria having immunostimulating ability, comprising a plant extract having an effect of enhancing the immunostimulating ability of bacteria having immunostimulating ability.

16. The enhancer according to claim 15, wherein the immunostimulatory activity is the ability to induce interferon α production in immune cells.

Images