Immune-activating composition and immune activation method

Abstract

To provide a novel immune-activating composition.SOLUTION: An immune-activating composition contains bacteria having immune-activating capability, and a plant extract having an action of suppressing immune-activating capability of the bacteria.SELECTED DRAWING: None

Description

[Technical Field]

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

[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 with 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 with 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, 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. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2012 / 091081 [Patent Document 2] Japanese Patent Application Laid-Open No. 2016-005452 [Patent Document 3] Japanese Patent Application Publication No. 2017-085975 Summary of the Invention

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

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

[0008] The present invention provides, for example, the following inventions. [1] Bacteria with immunostimulatory properties and plant extracts with inhibitory effects on the immunostimulatory properties of the above bacteria An immunostimulatory composition comprising: [2] Bacteria with immunostimulatory properties and plant extracts with an inhibitory effect on the immunostimulatory properties of the above bacteria A method for immunostimulation, comprising administering or ingesting the above to a subject. [3] Bacteria with immunostimulatory properties and plant extracts with an effect of suppressing the immunostimulatory properties of the above bacteria A method for suppressing the immunostimulatory effect induced by bacteria having immunostimulatory ability, comprising administering or ingesting the above to a subject. [4] A suppressor of immunostimulatory activity induced by bacteria having immunostimulatory activity, comprising a plant extract having an effect of suppressing the immunostimulatory activity of bacteria having immunostimulatory activity. [5] Bacteria with immunostimulatory properties and plant extracts with an effect of suppressing the immunostimulatory properties of the above bacteria 10. An agent for suppressing immunostimulatory activity induced by bacteria having immunostimulatory activity, comprising: [6] An immunostimulatory composition induced by bacteria having immunostimulatory ability, comprising a plant extract having an effect of suppressing the immunostimulatory ability of bacteria having immunostimulatory ability. [7] For producing an immunostimulatory composition, Use of bacteria having immunostimulatory activity and a plant extract having an effect of suppressing the immunostimulatory activity of the bacteria. [8] Use of a plant extract having an effect of suppressing the immunostimulating ability of a bacterium having immunostimulating ability for producing an immunostimulating composition induced by the bacterium. [9] Use of a plant extract having an effect of suppressing the immunostimulating ability of a bacterium having immunostimulating ability for producing an immunostimulating composition to be administered or ingested in combination with the bacterium.

[10] Use of bacteria having immunostimulatory ability for producing a composition for immunostimulation, which is administered or ingested in combination with a plant extract having an effect of suppressing the immunostimulatory ability of bacteria having immunostimulatory ability.

[11] A plant extract having an effect of suppressing the immunostimulatory activity of bacteria having immunostimulatory activity, for use in immunostimulation induced by the bacteria.

[12] A bacterium having immunostimulatory activity for use in a therapeutic method for immunostimulation, which is administered or ingested in combination with a plant extract having an effect of suppressing the immunostimulatory activity of the bacterium.

[13] A plant extract having an effect of suppressing the immunostimulatory activity of bacteria having immunostimulatory activity, for use in a therapeutic method for immunostimulation, wherein the plant extract is administered or ingested in combination with bacteria having immunostimulatory activity.

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

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

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

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

[17] The composition, method, inhibitor, use, plant extract or bacterium described in

[16] , wherein the lactic acid bacteria are Lactococcus bacteria.

[18] The composition, method, inhibitor, use, plant extract or bacterium described in

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

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

[10] and

[16] to

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

[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 immunostimulating composition.

[21] A composition, method, inhibitor, use, plant extract or bacterium described in any of [1] to

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

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

[22] , wherein the immune cells are pDCs.

[24] A composition, method, inhibitor, use, plant extract or bacterium described in any of [1] to

[23] , wherein the immunostimulatory activity suppression effect suppresses the induction of interferon α production in the immune cells to 0.10 times or more but less than 1.00 times compared to an immunostimulatory composition containing only bacteria having immunostimulatory activity.

[25] The composition, method, inhibitor, use, plant extract or bacterium described in any of [1] to

[24] , wherein the plant extract is one or more plant extracts selected from the group consisting of kale extract, Garcinia extract, Rahmania extract, and Bay extract.

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

[27] A method for producing an immunostimulating composition, comprising adding to a bacterium having immunostimulating ability a plant extract having an effect of suppressing the immunostimulating ability of the bacterium. DETAILED DESCRIPTION OF THE INVENTION

[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. 2024-072362, 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 possible combinations of one element or two or more elements constituting the group, and may be, for example, one of the elements constituting the group, or a combination of any two, three, four, five, six, seven, eight, nine or more elements constituting the group.

[0014] [Immunostimulatory composition] A first embodiment of the present disclosure relates to a composition for immunostimulation. The composition for immunostimulation includes bacteria having immunostimulatory activity and a plant extract having an effect of suppressing immunostimulatory activity. The composition for immunostimulation according to one embodiment includes bacteria having immunostimulatory activity, 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 the 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 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 the effect of increasing the amount of cytokine expression by dendritic cells when compared to when the immunostimulatory bacteria are not administered, ingested, etc. The immunostimulatory effect (immunostimulatory ability) according to one embodiment may include plasmacytoid dendritic cell activation effect (pDC activation effect) (plasmacytoid dendritic cell activation ability (pDC activation ability)), or may be 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 the effect of increasing (ability to increase) the amount of cytokine expression by pDC when compared to when the immunostimulatory bacteria are not administered, ingested, etc.

[0017] Plasmacytoid dendritic cells (pDCs), 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 other pathogens. Representative type I interferons are interferon α (IFN-α) and interferon β (IFN-β), and not only these but also type II interferon, interferon λ (IFN-λ), is thought to have immunostimulatory effects. 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, National Patent Microorganisms Depositary (NPMD, Japan) under accession number FERM BP-10914.

[0018] In one embodiment, the immunostimulatory effect (immunostimulatory ability), dendritic cell activation effect (dendritic cell activation ability), or pDC activation effect (pDC activation ability) may include an effect of promoting the production of at least one IFN (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 immunostimulating activity 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 immunostimulating activity. 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 immunostimulating activity. The function of immune cells may be evaluated, for example, using as an index an activation marker molecule (protein, etc.) that indicates that immune cells are activated, or a molecule (protein, etc.; e.g., IFN, etc.) produced when immune cells are activated.

[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 according to the following procedures (i) to (iv). (i) Mouse-derived bone marrow cells from which red blood cells had been removed were added to RPMI medium prepared so that the final concentrations of each component listed in the <Culture of Medium> below were the same as those listed in the <Culture of Medium> below, at a concentration of 1 × 10 6 The cells are suspended at a concentration of 1000 cells / mL to prepare a cell suspension. <Culture composition> 10% by volume FBS 100U / mL penicillin / streptomycin 1mM sodium pyruvate 2.5mM HEPES ·1 mass% non-essential amino acids (NEAA) for MEM 50 μM β-mercaptoethanol 100ng / mL Flt-3L (ii) 1 mL of the prepared cell suspension is seeded into each well and cultured in a CO2 incubator at 37°C and 5% CO2 by volume for 1 week to induce pDCs. (iii) 2 × 10 bone marrow cells containing induced pDCs 5 The bacteria are suspended at a concentration of bacteria / mL, and 200 μL of each are seeded onto a 96-well plate, and 2 μL of a composition containing bacteria adjusted to a concentration of 1 mg / mL with PBS is added to each well. (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 the production of at least one IFN selected from the group consisting of IFN-α, IFN-β, and IFN-λ, 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 enzymes, 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 it may be determined that the bacteria have immunostimulatory activity if the function of immune cells is enhanced when the cells are cultured in the presence of the bacteria having immunostimulatory activity compared to when the 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 combining the bacteria at a final concentration of 10 μg / mL with pDCs collected from mouse bone marrow and cultured for 7 days in a cell culture medium containing 100 ng / mL Flt3-L at a final concentration of 2 × 10 5 By co-culturing with bone marrow cells at 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, Pediococcus, Streptococcus, Enterococcus, Lactobacillus, Bacillus, and Heyndrickxia (formerly classified as Bacillus).

[0026] The Lactobacillus bacteria of the present invention include bacteria that were classified into the genus Lactobacillus before the reclassification of the genus Lactobacillus. For example, with the reclassification of the Lactobacillus genus, the following new species have been added: Acetilactobacillus, Agrilactobacillus, Amylolactobacillus, Apilactobacillus, Bombilactobacillus, Companilactobacillus, Dellaglioa, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, Lacticaseibacillus, Lactiplantibacillus, and Lapidilactobacillus. This includes bacteria classified into the genera Lapidilactobacillus, Latilactobacillus, Lentilactobacillus, Levilactobacillus, Ligilactobacillus, Limosilactobacillus, Liquorilactobacillus, Loigolactobacillus, Paralactobacillus, Paucilactobacillus, Schleiferilactobacillus, and Secundilactobacillus.

[0027] Among the above, preferred bacteria are those of the genus Oenococcus, Bifidobacterium, Lentilactobacillus, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, Lactobacillus, Lactiplantibacillus, Lacticaseibacillus, and Limosilactobacillus, with Lactococcus being 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] Examples of the Bifidobacterium genus include, but are not limited to, Bifidobacterium animalis subsp. lactis and Bifidobacterium longum subsp. infantis. Specific examples of Bifidobacterium genus include Bifidobacterium animalis subsp. lactis JCM10602, Bifidobacterium longum subsp. infantis JCM1222, and Bifidobacterium longum subsp. longum BB536.

[0030] The Weissella genus bacteria include, but are not limited to, Weissella paramesenteroides and Weissella viridescens. Specific examples of Weissella genus bacteria include Weissella paramesenteroides JCM9890 and Weissella viridescens JCM1174.

[0031] The 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] Examples of the Lactococcus bacteria include, but are not limited to, Lactococcus lactis, Lactococcus lactis subsp. lactis, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. hordniae, Lactococcus curvatus, and Lactococcus plantarum.

[0033] Specific examples of the above-mentioned Lactococcus genus 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 27861, Lactococcus lactis subsp. 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 are not particularly limited, but include, for example, Leuconostoc carnosum and Leuconostoc lactis, etc. Specific examples of Leuconostoc genus bacteria include Leuconostoc carnosum JCM9695 and Leuconostoc lactis NBRC12455, etc.

[0035] Examples of the Pediococcus bacteria include, but are not limited to, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus cellicola, Pediococcus claussenii, Pediococcus damnosus, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, and Pediococcus stilesii. Specific examples of bacteria of the genus Pediococcus include Pediococcus acidilactici JCM8797, Pediococcus acidilactici K15, and Pediococcus damnosus JCM5886.

[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 include, for example, 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, Lacticaseibacillus paracasei subsp. paracasei, Lacticaseibacillus delbrueckii, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fructivorans, Lactobacillus hilgardii, and Lactobacillus rhamnosus. rhamnosus, Lacticaseibacillus rhamnosus, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus parakefiri, Lentilactobacillus parakefiri, Lactobacillus plantarum, Lactiplantibacillus plantarum, Lactobacillus plantarum subsp. plantarum plantarum), Lactiplantibacillus plantarum subsp. plantarumExamples include Lactobacillus plantarum, Lactobacillus pentosus, Lactiplantibacillus pentosus, Lactobacillus helveticus, Limosilactobacillus reuteri subsp. reuteri, Lactobacillus crispatus, Lactobacillus 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 Lactiplantibacillus plantarum) L-137, Lactobacillus pentosus (newly classified as Lactiplantibacillus pentosus) ONRICb0240, Lactobacillus paraplantarum (newly classified as Lactiplantibacillus paraplantarum) JCM1149, Lactobacillus acidophilus JCM1021, Lactobacillus acidophilus JCM1132, Lactobacillus helveticus JCM1003, 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, for example, Bacillus coagulans SANK70258 strain, etc.

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

[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. Gluconacetobacter is preferred, Gluconacetobacter hansenii is more preferred, and Gluconacetobacter hansenii GK-1 is even more preferred.

[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 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, Bacteroides uniformis JCM13286, and Bacteroides uniformis JCM13287 and Bacteroides uniformis JCM13288.

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

[0047] Specific examples of bacteria 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 bacteria 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 celereculescens 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 a preferred embodiment, the bacterium may be at least one bacterium selected from the group consisting of Lactobacillus rhamnosus CRL1505 or a mutant thereof, Gluconacetobacter hansenii or a mutant thereof, Lactobacillus acidophilus (GK-1) L-92 or a mutant thereof, Lactobacillus plantarum L-137 or a mutant thereof, Lactobacillus bulgaricus OLL1073R-1 or a mutant thereof, and Lactococcus lactis subsp. lactis JCM5805 or a mutant thereof.

[0056] Of the above lactic acid bacteria and acetic acid bacteria, JCM strains can be obtained from the Microbial Materials Development Laboratory, RIKEN BioResource Center (3-1-1 Takanodai, Tsukuba, Ibaraki Prefecture, Japan); NBRC strains can be obtained from the Biological Genetic Resources Division, National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu, Chiba Prefecture, Japan); NRIC strains can be obtained from the Tokyo University of Agriculture and Technology Culture Collection (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo, Japan); and ATCC strains 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 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 Engineering Division of the RIKEN BioResource Center, as described above. However, in the present invention, the same strain as the JCM strain can be used that is preserved in a collection institution other than the RIKEN BioResource Center. 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, Chiba Prefecture), the Tokyo University of Agriculture, Culture Collection (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), and the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia, USA). 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 strain and has immunostimulatory ability equivalent to or greater than that of the above-mentioned strain. Whether a mutant strain has immunostimulatory ability equivalent to or greater than that of the above-mentioned 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 treating the bacterium with a mutagen such as ultraviolet (UV) or a DNA alkylating agent, or by introducing mutations into the strains using known genetic engineering techniques 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 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 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 8More than 1×10 14 pcs or less, 1×10 8 More than 1×10 13 pcs or less, 1×10 8 More than 1×10 12 pcs or less, 1×10 9 More than 1×10 14 pcs or less, 1×10 9 More than 1×10 13 pcs or less, 1×10 10 More than 1×10 14 pcs or less, 1×10 10 More than 1×10 13 10 or less, or 1 x 10 10 More than 1×10 12 The number of bacteria 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 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 bacteria 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 respectively. For example, with respect to the total mass of the immunostimulating composition of the present embodiment, the dry mass of the bacterium 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 50% 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 5% 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 6% by mass or less, 0.02% by mass or more and 50% by mass or less, 0.0.02% 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 and above. Less than 15% by mass, 0.10% to 100% by mass but not fully expressed, 0.10% to 95% by mass, 0.10% to 90% by mass, 0.10% to 80% by mass, 0.10% to 70% by mass, 0.10% to 60% by mass, 0.10% to 50% by mass, 0.10% to 30% by mass, 0.10% to 25% by mass, 0.10% to 20% by mass, 0.10% to 15% by mass, 0.30% to 100% by mass but not fully expressed, 0.30% to 95% by mass, 0.30% to 90% by mass 0.30% to 80% mass, 0.30% to 70% mass, 0.30% to 60% mass, 0.30% to 50% mass, 0.30% to 30% mass, 0.30% to 25% mass, 0.30% to 20% mass, 0.30% to 15% mass, 1.0% to 100% mass (not fully expressed), 1.0% to 95% mass, 1.0% to 90% mass, 1.0% to 80% mass, 1.0% to 70% mass, 1.0% to 60% mass, 1.0% to 50% mass Below 1%, 1.0% to 30% of mass, 1.0% to 25% of mass, 1.0% to 20% of mass, 1.0% to 15% of mass, 1.5% to 100% of mass not fully expressed, 1.5% to 95% of mass, 1.5% to 90% of mass, 1.5% to 80% of mass, 1.5% to 70% of mass, 1.5% to 60% of mass, 1.5% to 50% of mass, 1.5% to 30% of mass, 1.5% to 25% of mass, 1.5% to 20% of mass, 1.5% to 15% of mass, 2.0% or more, 100% or less; 2.0% or more, 95% or less; 2.0% or more, 90% or less; 2.0% or more, 80% or less; 2.0% or more, 70% or less; 2.0% or more, 60% or less; 2.0% or more, 50% or less; 2.0% or more, 30% or less; 2.0% or more, 25% or less; 2.0% or more, 20% or less; 2.0% or more, 15% or less; 2.5% or more, 100% or less; 2.5% or more, 95% or less; 2.5% or more, 90% or less; 2.5% or more, 80% or less; 2.5% or less... 70% or less by mass, 2.5% or more by mass but less than 60% by mass, 2.5% or more by mass but less than 50% by mass, 2.5% or more by mass but less than 30% by mass, 2.5% or more by mass but less than 25% by mass, 2.5% or more by mass but less than 20% by mass, 2.5% or more by mass but less than 15% by mass, 3.0% or more by mass but less than 100% by mass, 3.0% or more by mass but less than 95% by mass, 3.0% or more by mass but less than 90% by mass, 3.0% or more by mass but less than 80% by mass, 3.0% or more by mass but less than 70% by mass, 3.0% or more by mass but less than 60% by mass, 3.0% or more by mass but less than 50% by mass, 3.0% or more by mass but less than 30% by mass, 3.0% or more by mass but less than 25% by mass. The following are categories: 3.0% to 20% mass, 3.0% to 15% mass, 5.0% to 100% mass (not fully expressed), 5.0% to 95% mass, 5.0% to 90% mass, 5.0% to 80% mass, 5.0% to 70% mass, 5.0% to 60% mass, 5.0% to 50% mass, 5.0% to 30% mass, 5.0% to 25% mass, 5.0% to 20% mass, 5.0% to 15% mass, 7.0% to 100% mass (not fully expressed), 7.0% to 95% mass, 7. 0% quality or higher, 90% quality or lower; 7.0% quality or higher, 80% quality or lower; 7.0% quality or higher, 70% quality or lower; 7.0% quality or higher, 60% quality or lower; 7.0% quality or higher, 50% quality or lower; 7.0% quality or higher, 30% quality or lower; 7.0% quality or higher, 25% quality or lower; 7.0% quality or higher, 20% quality or lower; 7.0% quality or higher, 15% quality or lower; 10.0% quality or higher, 100% quality or lower; 10.0% quality or higher, 95% quality or lower; 10.0% quality or higher, 90% quality or lower; 10.0% quality or higher, 80% quality or lower; 10.0% quality or higher, 70% quality or lower; 10.0% quality or higher, 60% quality or lower; 10.The dry mass of the bacterium according to this embodiment may be 0% to 50% by mass, 10.0% to 30% by mass, 10.0% to 25% by mass, 10.0% to 20% by mass, or 10.0% to 15% by mass. 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 determined 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, 1.0×10 4 Cells / mL or more, 1.0×10 5 Cells / mL or more, 1.0×10 6 Cells / mL or more, 1.0×10 7 cells / mL or 4.0 x 10 7 cells / mL or greater, and may be 1.0 x 10 11 Cells / mL or less, 1.0×10 10 Cells / mL or less, 3.0×10 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 1.0×10 11 Cells / mL or less, 1.0×10 3 cells / mL or more 1.0×10 10 Cells / mL or less, 1.0×10 3 cells / mL or more 3.0×10 9 Cells / mL or less, 1.0×10 3 cells / mL or more 1.0×10 9 Cells / mL or less, 1.0×10 4 cells / mL or more 1.0×10 11 Cells / mL or less, 1.0×10 4 cells / mL or more 1.0×10 10 Cells / mL or less, 1.0×10 4 cells / mL or more 3.0×10 9Cells / mL or less, 1.0×10 4 cells / mL or more 1.0×10 9 Cells / mL or less, 1.0×10 5 cells / mL or more 1.0×10 11 Cells / mL or less, 1.0×10 5 cells / mL or more 1.0×10 10 Cells / mL or less, 1.0×10 5 cells / mL or more 3.0×10 9 Cells / mL or less, 1.0×10 5 cells / mL or more 1.0×10 9 Cells / mL or less, 1.0×10 6 cells / mL or more 1.0×10 11 Cells / mL or less, 1.0×10 6 cells / mL or more 1.0×10 10 Cells / mL or less, 1.0×10 6 cells / mL or more 3.0×10 9 Cells / mL or less, 1.0×10 7 cells / mL or more 1.0×10 11 Cells / mL or less, 1.0×10 7 cells / mL or more 1.0×10 10 Cells / mL or less, 1.0×10 7 cells / mL or more 1.0×10 9 Cells / mL or less, 4.0×10 7 cells / mL or more 1.0×10 11 Cells / mL or less, 4.0×10 7 cells / mL or more 1.0×10 10 Cells / mL or less, 4.0×10 7 cells / mL or more 1.0×10 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, 3.0×108 pcs 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 x 10 or more, 14 pcs or less, 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 pcs or less, 1.0×10 7 pcs or more 1.0×10 13 pcs or less, 1.0×10 7 pcs or more 1.0×10 12 pcs or less, 1.0×10 8 pcs or more 1.0×10 14 pcs or less, 1.0×10 8 pcs or more 1.0×10 13 pcs or less, 1.0×10 8 pcs or more 1.0×10 12 pcs or less, 3.0×10 8 pcs or more 1.0×10 14 pcs or less, 3.0×10 8 pcs or more 1.0×10 13 pcs or less, 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 pcs or less, 1.0×10 9 pcs or more 1.0×10 14 pcs or less, 1.0×10 9 pcs or more 1.0×10 13 pcs or less, 1.0×10 9 pcs or more 1.0×10 12 Less than or equal to 5.0×109 pcs 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 pcs or less, 1.0×10 10 pcs or more 1.0×10 14 pcs or less, 1.0×10 10 pcs or more 1.0×10 13 pcs or less, 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 pcs or more 5.0×10 13 Less than or equal to 5.0×10 10 pcs or more 1.0×10 12 pcs or less, 1.0×10 11 pcs or more 1.0×10 14 pcs or less, 1.0×10 11 pcs or more 1.0×10 13 pcs or less, 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 suppressing the immunostimulatory activity of immunostimulatory bacteria" refers to a plant extract that has an effect of suppressing the immunostimulatory activity of immunostimulatory bacteria, and may be a plant extract that, when co-cultured with the immunostimulatory bacteria and immune cells, has a lower 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 suppressing the immunostimulatory ability of bacteria is not particularly limited, and if the function of immune cells in the presence of bacteria having immunostimulatory ability is suppressed 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 suppressing 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 suppressing the immunostimulatory ability of bacteria having immunostimulatory ability can be confirmed by a decrease in 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') Mouse-derived bone marrow cells from which red blood cells had been removed were added to RPMI medium prepared so that the components listed below (Culture medium composition) were added to the final concentrations listed below (Culture medium composition) at a concentration of 1 × 10 6 The cells are suspended at a concentration of 1000 cells / mL to prepare a cell suspension. <Culture composition> 10% by volume FBS 100U / mL penicillin / streptomycin 1mM sodium pyruvate 2.5mM HEPES ·1 mass% non-essential amino acids (NEAA) for MEM 50 μM β-mercaptoethanol 100ng / mL Flt-3L (ii') 1 mL of the prepared cell suspension is seeded into each well and cultured in a CO2 incubator at 37°C and 5% CO2 by volume for 1 week to induce pDCs. (iii') 2 × 10 bone marrow cells containing induced pDCs 5 The bacteria are suspended at a concentration of bacteria / mL, and 200 μL of each is seeded into a 96-well plate. 2 μL of a bacterial suspension adjusted to a concentration of 1 mg / mL with PBS is added to each well, and then a plant extract solution adjusted to a predetermined concentration 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.

[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, 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, carob extract, etc.), and extracts of plants belonging to the Asteraceae family. The extract may be an extract of a plant belonging to the Zingiberaceae family (e.g., ginger extract), an extract of a plant belonging to the Cannabaceae family (e.g., hop extract), an extract of a plant belonging to the Araliaceae family (e.g., red ginseng extract), an extract of a plant belonging to the Clusiaceae family (e.g., Garcinia extract), an extract of a plant belonging to the Lamiaceae family (e.g., Rahma extract), or an extract of a plant belonging to the Lauraceae family (e.g., Bay extract).

[0072] Preferred plant extracts having immunosuppressive effects are plant extracts belonging to the Bromeliaceae family, plant extracts belonging to the Hypericaceae family, plant extracts belonging to the Brassicaceae family (e.g., kale extract, etc.), plant extracts belonging to the Poaceae family, plant extracts belonging to the Fabaceae family, plant extracts belonging to the Asteraceae family, plant extracts belonging to the Zingiberaceae family, plant extracts belonging to the Cannabaceae family, plant extracts belonging to the Araliaceae family, plant extracts belonging to the Garcinia family (Clusiaceae) (e.g., Garcinia extract, etc.), plant extracts belonging to the Lamiaceae family (Lamiaceae) (e.g., Rahma extract, etc.), and plant extracts belonging to the Lauraceae family (Lauraceae) (e.g., Bay extract, etc.).

[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 serrata). 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] Preferred plant extracts having immunosuppressive activity include extracts of plants belonging to the genus Ananas, extracts of plants belonging to the genus Garcinia, extracts of plants belonging to the genus Lepidium, extracts of plants belonging to the genus Oryza, extracts of plants belonging to the genus Glycine, extracts of plants belonging to the genus Hypericum, extracts of plants belonging to the genus Chrysanthemum, extracts of plants belonging to the genus Ceratonia, extracts of plants belonging to the genus Zingiber, extracts of plants belonging to the genus Humulus, extracts of plants belonging to the genus Panax, extracts of plants belonging to the genus Brassica (e.g., kale extract, etc.), extracts of plants belonging to Garcinia (e.g., Garcinia extract), extracts of plants belonging to the genus Melissa (e.g., Rhaematous rhododendron extract, etc.), and extracts of plants belonging to the genus Laurus (e.g., bay laurel extract, etc.).

[0075] Examples of plant extracts 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] The plant extract having an immunosuppressive effect may preferably be a plant extract used as an agent for improving skin barrier function, a plant extract used as an agent for recovering from fatigue (e.g., kale extract, etc.), a plant extract used as an anti-aging agent (e.g., kale extract, etc.), a plant extract used as an agent for improving menopausal disorders, a plant extract used as an agent for preventing arteriosclerosis (e.g., kale extract, etc.), a plant extract used as an anti-inflammatory agent (e.g., Garcinia extract and Bay extract, etc.), a plant extract used as an immunostimulant (e.g., kale extract, etc.), a plant extract used as an antibacterial agent, a plant extract used as an agent for reducing body fat (e.g., Garcinia extract, etc.), a plant extract used as an agent for improving brain function, etc.

[0077] Examples of plant extracts 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] As the plant extract having an immunosuppressive effect, preferred are plant extracts used as bittering agents, plant extracts used as antioxidants (e.g., kale extract, Garcinia extract, etc.), plant extracts used as sweeteners (e.g., Garcinia extract, etc.), and plant extracts used as anti-glycation agents (e.g., herb extracts, etc.), and more preferred are plant extracts used as antioxidants (e.g., kale extract, Garcinia extract, etc.) and plant extracts used as sweeteners (e.g., Garcinia extract, 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, Rafuma extract, and bay extract, etc., and is preferably one or more plant extracts selected from the group consisting of herb extract, kale extract, Garcinia extract, Rafuma extract, and bay extract, and more preferably one or more plant extracts selected from the group consisting of kale extract, Garcinia extract, Rafuma extract, and bay extract.

[0080] The pineapple extract is preferably a pineapple extract, which may be in liquid or solid form such as powder, and may be a fat-soluble component extracted by heating and refluxing pineapple in an ethanol solution of 80% to 100% by volume, 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 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 can be in liquid form or solid form such as powder. The maca extract can 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 can be used.

[0083] The fermented extract of rice germ and soybeans can be in liquid form or in solid form such as powder. The extract can be prepared by fermenting rice germ and soybeans with lactic acid bacteria and then extracting them 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. can be used.

[0084] St. John's wort (Hypericum perforatum) extract can be used in liquid or solid form such as powder, and 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 liquid or solid form such as powder, and may be 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.

[0086] The herbal extract can be in liquid form or in solid form such as 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 liquid form or in solid form such as 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 acetic acid ethyl ester; 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). They can also be used as extracts obtained by supercritical extraction using supercritical carbon dioxide or the like. From the perspective of odor removal, the extraction temperature with 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 liquid or solid form such as 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 can be in liquid or solid form such as powder, and can 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 can 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 Laboratory can be used.

[0095] The inhibitory effect of the plant extract according to this embodiment on the immunostimulatory activity 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 preferably at least 0.10-fold, at least 0.15-fold, at least 0.20-fold, at least 0.30-fold, or at least 0.40-fold higher than that in the absence of the plant extract, as a lower limit, and preferably at most 1.0-fold, at most 0.95-fold, at most 0.90-fold, at most 0.85-fold, at most 0.80-fold, at most 0.75-fold, at most 0.70-fold, at most 0.65-fold, at most 0.60-fold, at most 0.55-fold, at most 0.50-fold, at most 0.45-fold, at most 0.40-fold, at most 0.35-fold, at most 0.30-fold, at most 0.25-fold, or at most 0.20-fold higher than that in the absence of the plant extract. These upper and lower limits can be arbitrarily combined. The effect of the plant extract according to this embodiment in suppressing the immunostimulatory activity of bacteria is, for example, when the IFN-α concentration measured by the above steps (i') to (iv') in the presence of the plant extract is greater than or equal to 0.10 times but less than 1.0 times, greater than or equal to 0.10 times but less than 0.95 times, greater than or equal to 0.10 times but less than 0.90 times, greater than or equal to 0.10 times but less than 0.85 times, or greater than or equal to 0.10 times but less than 0.10 times, compared to the case in the absence of the plant extract. 0.80 times or more, 0.10 times or more and 0.75 times or less, 0.10 times or more and 0.70 times or less, 0.10 times or more and 0.65 times or less, 0.10 times or more and 0.60 times or less, 0.10 times or more and 0.55 0.10 times or more and 0.50 times or less, 0.10 times or more and 0.45 times or less, 0.10 times or more and 0.40 times or less, 0.10 times or more and 0.35 times or less, 0.10 times or more and 0.30 times or less, 0.1 0x or more and 0.25x or less, 0.10x or more and 0.20x or less, 0.15x or more and less than 1.0x, 0.15x or more and 0.95x or less, 0.15x or more and 0.90x or less, 0.15x or more and 0.85x or less, 0.15x or more and 0.80x or less, 0.15x or more and 0.75x or less, 0.15x or more and 0.70x or less, 0.15x or more and 0.65x or less, 0.15x or more and 0.60x or less, 0. 15x or more and 0.55x or less, 0.15x or more and 0.50x or less, 0.15x or more and 0.45x or less, 0.15x or more and 0.40x or less, 0.15x or more and 0.35x or less, 0.15x or more and 0.30x or less, 0.15x or more and 0.25x or less, 0.15x or more and 0.20x or less, 0.20x or more and less than 1.0x, 0.20x or more and 0.95x or less, 0.20x or more and 0.90x or less, 0.20 times to 0.85 times, 0.20 times to 0.80 times, 0.20 times to 0.75 times, 0.20 times to 0.70 times, 0.20 times to 0.65 times , 0.20 times to 0.60 times, 0.20 times to 0.55 times, 0.20 times to 0.50 times, 0.20 times to 0.45 times, 0.20 times to 0.40 0.20 times or more and 0.35 times or less, 0.20 times or more and 0.30 times or less, 0.20 times or more and 0.25 times or less, 0.30 times or more and less than 1.0 times, 0.30 times or more and 0. 95x or less, 0.30x or more and 0.90x or less, 0.30x or more and 0.85x or less, 0.30x or more and 0.80x or less, 0.30x or more and 0.75x or less, 0.30x or more It is preferably 0.70 times or less, 0.30 times or more and 0.65 times or less, 0.30 times or more and 0.60 times or less, 0.30 times or more and 0.55 times or less, 0.30 times or more and 0.50 times or less, 0.30 times or more and 0.45 times or less, 0.30 times or more and 0.40 times or less, 0.30 times or more and 0.35 times or less, 0.40 times or more and 0.95 times or less, 0.40 times or more and 0.90 times or less, 0.40 times or more and 0.85 times or less, 0.40 times or more and 0.80 times or less, 0.40 times or more and 0.75 times or less, 0.40 times or more and 0.70 times or less, 0.40 times or more and 0.65 times or less, 0.40 times or more and 0.60 times or less, 0.40 times or more and 0.55 times or less, 0.40 times or more and 0.50 times or less, or 0.40 times or more and 0.45 times or less. .

[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. 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 or more and 90.0% by mass or less, 0.000001% by mass or more and 85.0% by mass or less, 0.000001% by mass or more and 80.0% by mass or less, 0.000001% by mass or more and 75.0% by mass or less, 0.000001% by mass or more and 50.0% by mass or less, 0.000001% by mass or more and 25.0% by mass or less, 0.000001% by mass or more and 10.0% by mass or less, 0.000001% by mass or more and 5.0% by mass or less, 0.000001% by mass or more and 1.0% by mass or less, 0.000001% by mass or more and 0.80% by mass or less, or 0.000001% by mass or more and 0.50% by mass or less, relative to the total amount of the immunostimulating composition. % 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, 0.00000 1 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 more and 80. 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, 0.0.00001% or more but less than 10.0% of mass, 0.00001% or more but less than 5.0% of mass, 0.00001% or more but less than 1.0% of mass, 0.00001% or more but less than 0.80% of mass, 0.00001% or more but less than 0.50% of mass, 0.00001% or more but less than 0.30% of mass, 0.00001% or more but less than 0.25% of mass, 0.00001% or more but less than 0.20% of mass, 0.00001% or more but less than 0.10% of mass, 0.00001% or more but less than 0.05% of mass, 0.00001% or more but less than 0.02% of mass, 0.00001% or more of mass 0.01% or less, 0.001% or more but less than 90.0% of mass, 0.001% or more but less than 85.0% of mass, 0.001% or more but less than 80.0% of mass, 0.001% or more but less than 75.0% of mass, 0.001% or more but less than 50.0% of mass, 0.001% or more but less than 25.0% of mass, 0.001% or more but less than 10.0% of mass, 0.001% or more but less than 5.0% of mass, 0.001% or more but less than 1.0% of mass, 0.001% or more but less than 0.80% of mass, 0.001% or more but less than 0.50% of mass, 0.001% or more but less than 0.30% of mass, 0 0.001% or more and less than 0.25% of mass, 0.001% or more and less than 0.20% of mass, 0.001% or more and less than 0.10% of mass, 0.001% or more and less than 0.05% of mass, 0.001% or more and less than 0.02% of mass, 0.001% or more and less than 0.01% of mass, 0.01% or more and less than 90.0% of mass, 0.01% or more and less than 85.0% of mass, 0.01% or more and less than 80.0% of mass, 0.01% or more and less than 75.0% of mass, 0.01% or more and less than 50.0% of mass, 0.01% or more and less than 25.0% of mass, 0.01% or more and more than 10.0% of mass. Below, 0.01% of mass and below 5.0% of mass, 0.01% of mass and below 1.0% of mass, 0.01% of mass and below 0.80% of mass, 0.01% of mass and below 0.50% of mass, 0.01% of mass and below 0.30% of mass, 0.01% of mass and below 0.25% of mass, 0.01% of mass and below 0.20% of mass, 0.01% of mass and below 0.10% of mass, 0.01% of mass and below 0.05% of mass, 0.01% of mass and below 0.02% of mass, 0.02% of mass and below 90.0% of mass, 0.02% of mass and below 85.0% of mass, 0.02% of mass and below 80.0% of mass, 0.0.02% or more, 75.0% or less; 0.02% or more, 50.0% or less; 0.02% or more, 25.0% or less; 0.02% or more, 10.0% or less; 0.02% or more, 5.0% or less; 0.02% or more, 1.0% or less; 0.02% or more, 0.8% or less; 0.02% or more, 0.5% or less; 0.02% or more, 0.3% or less; 0.02% or more, 0.25% or less; 0.02% or more, 0.2% or less; 0.02% or more, 0.10% or less; 0.02% or more, 0.05% or less; 0.0 5% or more, 90.0% or less; 0.05% or more, 85.0% or less; 0.05% or more, 80.0% or less; 0.05% or more, 75.0% or less; 0.05% or more, 50.0% or less; 0.05% or more, 25.0% or less; 0.05% or more, 10.0% or less; 0.05% or more, 5.0% or less; 0.05% or more, 1.0% or less; 0.05% or more, 0.8% or less; 0.05% or more, 0.5% or less; 0.05% or more, 0.3% or less; 0.05% or more, 0.25% or less; 0.05% or more, 0.05% or less. Quantity % or more but less than 0.20% of mass, 0.05% or more but less than 0.10% of mass, 0.10% or more but less than 90.0% of mass, 0.10% or more but less than 85.0% of mass, 0.10% or more but less than 80.0% of mass, 0.10% or more but less than 75.0% of mass, 0.10% or more but less than 50.0% of mass, 0.10% or more but less than 25.0% of mass, 0.10% or more but less than 10.0% of mass, 0.10% or more but less than 5.0% of mass, 0.10% or more but less than 1.0% of mass, 0.10% or more but less than 0.80% of mass, 0.10% or more but less than 0.50% of mass, 0.10% of mass Above 0.30% by mass, 0.10% by mass and below 0.25% by mass, 0.10% by mass and below 0.20% by mass, 0.50% by mass and below 90.0% by mass, 0.50% by mass and below 85.0% by mass, 0.50% by mass and below 80.0% by mass, 0.50% by mass and below 75.0% by mass, 0.50% by mass and below 50.0% by mass, 0.50% by mass and below 25.0% by mass, 0.50% by mass and below 10.0% by mass, 0.50% by mass and below 5.0% by mass, 0.50% by mass and below 1.0% by mass, 0.50% by mass and below 0.80% by mass, 1% by mass and below 90%.0% or less, 1% or more but less than 85.0%, 1% or more but less than 80.0%, 1% or more but less than 75.0%, 1% or more but less than 50.0%, 1% or more but less than 25.0%, 1% or more but less than 10.0%, 1% or more but less than 5.0%, 2% or more but less than 90.0%, 2% or more but less than 85.0%, 2% or more but less than 80.0%, 2% or more but less than 75.0%, 2% or more but less than 50.0%, 2% or more but less than 25.0%, 2% or more but less than 10.0%, 2% or more but less than 5.0%

[0097] The ratio of the content (% by mass) of immunostimulatory bacteria to the content (% by mass) of plant extract in an immunostimulatory composition according to one embodiment ((content of immunostimulatory bacteria) / (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 composition for immunostimulation 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, 0.001 or more and 1,000 or less, 50 or more, 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, 0.00 3 to 50000, 0.003 to 10000, 0.003 to 1000, 0.003 to 500, 0.003 to 100, 0.003 to 50, 0.003 to 10 Below, 0.003 to 5, 0.003 to 1.0, 0.003 to 0.75, 0.003 to 0.5, 0.003 to 0.3, 0.005 to 50000, 0.005 10000 or more, 0.005 or more, 1000 or less, 0.005 or more, 500 or less, 0.005 or more, 100 or less, 0.005 or more, 50 or less, 0.005 or more, 10 or less, 0.005 or more, 5 or less, 0. 005 to 1.0, 0.005 to 0.75, 0.005 to 0.5, 0.005 to 0.3, 0.01 to 50000, 0.01 to 10000, 0.01 to 10 00 or less, 0.01 to 500, 0.01 to 100, 0.01 to 50, 0.01 to 10, 0.01 to 5, 0.01 to 1.0, 0.01 to 0.75, 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, 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, 0.015 0.5 or more, 0.015 or more and 0.3 or less, 0.02 or more and 50,000 or more, 0.02 or more and 10,000 or more, 0.02 or more and 1000 or less, 0.02 or more and 500 or less, 0.02 or more Above 100, 0.02 to 50, 0.02 to 10, 0.02 to 5, 0.02 to 1.0, 0.02 to 0.75, 0.02 to 0.5, 0.02 0.3 or more, 0.025 or more and 50,000 or less, 0.025 or more and 10,000 or more, 0.025 or more and 1,000 or more, 0.025 or more and 500 or less, 0.025 or more and 100 or less, 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, 0.025 to 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 as 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., they 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 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 at least one week, 2 weeks or more, 1 month or more, 3 months or more, 6 months or more, 1 year or more, 3 years or more, 5 years or more, or 10 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 suppressing 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 suppressed immunostimulating activity, which is prepared by adding a plant extract having an effect of suppressing the immunostimulating activity of a bacterium having immunostimulating activity to the bacterium. The immunostimulating composition or the immunostimulating composition with suppressed immunostimulating activity is preferable from the viewpoint that the addition of the plant extract suppresses the immunostimulating activity of the bacterium.

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

[0107] In one embodiment, the inhibitor has an ability to induce interferon-α production in immune cells, preferably at least 0.10-fold, at least 0.15-fold, at least 0.20-fold, at least 0.30-fold, or at least 0.40-fold, as a lower limit, and preferably at most 1.00-fold, at most 0.95-fold, at most 0.90-fold, at most 0.85-fold, at most 0.80-fold, at most 0.75-fold, at most 0.70-fold, at most 0.65-fold, at most 0.60-fold, at most 0.55-fold, at most 0.50-fold, at most 0.45-fold, at most 0.40-fold, at most 0.35-fold, at most 0.30-fold, at most 0.25-fold, or at most 0.20-fold, as an upper limit, compared to an immunostimulatory composition containing only bacteria having immunostimulatory ability. These upper and lower limits can be arbitrarily combined, and may be the same as the ranges described for the bacterial immunostimulatory ability inhibitory effect of the plant extract according to this embodiment.

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

[0109] [Method for immunostimulation, method for suppressing immunostimulatory activity induced by bacteria having immunostimulatory ability] A third embodiment of the present disclosure relates to an immunostimulatory method and a method for suppressing immunostimulatory activity induced by bacteria having immunostimulatory activity (hereinafter also referred to simply as "suppression method"). The immunostimulatory method and the suppression method involve administering to or ingesting to a subject bacteria having immunostimulatory activity and a plant extract having an effect of suppressing the immunostimulatory activity of the bacteria. The suppression method according to this embodiment suppresses immunostimulatory activity compared to when only bacteria having immunostimulatory activity 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 may 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 suppressing 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 the 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 suppressed immunostimulatory activity, by adding a plant extract that has an effect of suppressing the immunostimulatory activity of a bacterium having immunostimulatory activity to the bacterium. The immunostimulatory composition or the immunostimulatory composition with suppressed immunostimulatory activity is preferable from the viewpoint that the addition of the plant extract suppresses the immunostimulatory activity of the bacterium.

[0114] Use of bacterial and plant extracts to prepare compositions A fifth embodiment of the present disclosure relates to the use of bacteria and a plant extract having an effect of suppressing 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 an effect of suppressing the immunostimulatory ability of the bacteria for producing a composition for immunostimulation. Another aspect of use is the use of a plant extract having an effect of suppressing 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 an effect of suppressing 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 extracts for use A sixth embodiment of the present disclosure relates to a plant extract having an effect of suppressing 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] Bacterial or plant extracts for use in therapeutic methods A seventh embodiment of the present disclosure relates to the use of a bacterium or a plant extract for use in a therapeutic method of immunostimulation. In one aspect, the bacterium has immunostimulatory activity for use in a therapeutic method of immunostimulation, and is administered or ingested in combination with a plant extract having an activity of suppressing the immunostimulatory activity of the bacterium. In another aspect, the plant extract has an activity of suppressing the immunostimulatory activity of a bacterium having immunostimulatory activity for use in a therapeutic method of immunostimulation, and is administered or ingested in combination with the bacterium having immunostimulatory activity. Specific aspects of the bacterium and plant extract 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, for example, be use for the purpose of or in conjunction with medical treatment. Use in a therapeutic method according to the present disclosure may, for example, involve 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 bacterial or plant extracts in non-therapeutic methods An eighth embodiment of the present disclosure relates to the use of bacteria or plant extracts for use in non-therapeutic immunostimulatory methods. In one aspect, the use relates to the use of bacteria having immunostimulatory activity in non-therapeutic immunostimulatory methods, where the bacteria are administered or ingested in combination with a plant extract having an activity of suppressing the immunostimulatory activity of the bacteria. In another aspect, the use relates to the use of a plant extract having an activity of suppressing the immunostimulatory activity of the bacteria in non-therapeutic immunostimulatory methods, where the bacteria are administered or ingested in combination with the bacteria having immunostimulatory activity. Specific aspects of the bacteria and plant extracts according to this embodiment are as described above. In this disclosure, "use in a non-therapeutic" method refers to the 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 be, for example, a use that is not intended for and / or does not involve medical treatment. Use in a non-therapeutic method according to the present disclosure may not involve, for example, a medical professional administering or ingesting the substance to a human or animal and / or instructing a human or animal to administer or ingest the substance. The use in the non-therapeutic method according to the present disclosure may be, for example, use for preventive purposes or health promotion purposes, and may be use for preventive purposes or health promotion purposes involving administration or ingestion of a pharmaceutical composition or a quasi-drug. The use in the non-therapeutic method according to the present disclosure may be, for example, use in healthy individuals. [Example]

[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 pDCs induced by JCM5805.

[0120] <1. Preparation of lactic acid bacteria suspension> JCM5805 cells were inoculated into 10 mL of MRS medium (Oxoid) and cultured for 24 hours at 30°C. Then, 1 mL of the cultured lactic acid bacteria cell suspension was inoculated into 100 mL of MRS medium and cultured for 24 hours at 30°C. 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 cells. The heat-killed cells 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 solution The pineapple extract is "Brightening Pine (registered trademark)" manufactured by Maruzen Pharmaceuticals, the mangosteen peel extract is "Mangosteen Aqua" manufactured by Nippon Shinyaku, the maca extract is "MACAXS (registered trademark)" manufactured by TOWA, the rice germ and fermented soybean extract is "Senspure (registered trademark)" manufactured by Toyo Hakko, the St. John's Wort (Hypericum perforatum) extract is "St. John's Wort" manufactured by FANCL, the chrysanthemum flower extract is "Chrysanthemum Flower Extract P" manufactured by Oryza Oil & Fat Chemical, and the herb (mixed herb of chamomile, Houttuynia cordata, hawthorn, and grape leaf) extract is " The plant extracts used were "AG Herb MIX (registered trademark)," "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 Rahma 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 a plant extract solution.

[0122] <3. Test Procedure> The test was carried out according to the following steps (i) to (iv). (i) Bone marrow cells derived from female BALB / c mice were cultured in RPMI medium (Sigma) containing the final concentrations of the components listed below (Culture medium composition) at 1 × 10 6 The cells were suspended at a density of 100 cells / mL to prepare a cell suspension. (Medium composition) 10% by volume FBS 100U / mL penicillin / streptomycin 1mM sodium pyruvate 2.5mM HEPES ·1 mass% non-essential amino acids (NEAA) for MEM 50 μM β-mercaptoethanol 100ng / mL Flt-3L (ii) 1 mL of the prepared cell suspension was seeded into each well and cultured in a CO2 incubator at 37°C and 5% CO2 for 1 week to induce pDCs. (iii) 2 × 10 bone marrow cells containing induced pDCs 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 to each well, and 5 μL of the plant extract solution (described above in 2) was simultaneously added. Similarly, the plant extract solution alone was added to a 96-well plate seeded with bone marrow cells containing pDCs. Similarly, the JCM5805 suspension alone was added to a 96-well plate seeded with bone marrow cells containing pDCs. (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 concentrations of each substance in the table refer to the final concentration.

[0124] [Table 1]

[0125] When any of the substances was used alone, it did not induce IFN-α production by pDCs. Kale extract, Garcinia cambogia peel extract, Rafuma leaf extract, and Bay leaf extract suppressed JCM5805-induced IFN-α production by pDCs, 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 pDCs.

Claims

1. An immunostimulatory composition comprising a bacterium having immunostimulatory ability and a plant extract having an effect of suppressing the immunostimulatory ability 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 immunostimulatory 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 with respect 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. 2. The immunostimulating composition according to claim 1, wherein a ratio of the content (mass%) of the bacteria having immunostimulatory ability to the content (mass%) of the plant extract ((content of plant extract) / (content of bacteria having immunostimulatory 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 an 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 ability suppressing effect is an effect of suppressing the immunostimulating ability to 0.10 times or more but less than 1.00 times.

11. The immunostimulatory composition according to claim 8 , wherein the plant extract is one or more plant extracts selected from the group consisting of kale extract, Garcinia extract, Rahmania extract, and Bay extract.

12. Bacteria with immunostimulatory activity and plant extracts with inhibitory effect on the immunostimulatory activity of said bacteria A method for immunostimulation, comprising administering or ingesting the above to a subject.

13. Bacteria with immunostimulatory activity and plant extracts with inhibitory effect on the immunostimulatory activity of said bacteria A method for suppressing the immunostimulatory effect induced by bacteria having immunostimulatory ability, comprising administering or ingesting the above to a subject.

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

15. An agent for suppressing immunostimulatory activity induced by bacteria having immunostimulatory activity, comprising a plant extract having an effect of suppressing the immunostimulatory activity of bacteria having immunostimulatory activity.

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

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