Novel lactic acid bacteria, novel lactic acid bacteria-containing composition, and method for producing same

Abstract

The present invention addresses the problem of providing bacteria which belong to Lactobacillus helveticus and which have an excellent IFN production inducing effect. Provided are bacteria that activate a plasmacytoid dendritic cells (pDC) and / or a myeloid dendritic cells (mDC) and that are capable of inducing I-type IFN production, said bacteria being classified as Lactobacillus helveticus and inducing I-type IFN production which is at least 1.5 times that of Lactobacillus helveticus SBT2171 (accession number: Ferm BP-5445).

Description

Novel lactic acid bacteria, composition containing novel lactic acid bacteria, and method for producing the same

[0001] The present invention relates to a novel bacterium belonging to the genus Lactobacillus, a composition containing the novel bacterium, a culture or treated cell product of the novel bacterium, and a method for producing the same.

[0002] Immunity is a biological defense mechanism present in many animals, responsible for eliminating pathogens such as bacteria and viruses that invade from the outside, as well as tumor cells that develop within the body. The immune system consists of two systems, the innate immune system and the adaptive immune system, which are responsible for the body's defense functions. The innate immune system is primarily responsible for the primary response to bacterial and viral infections, and dendritic cells are a powerful and important component of the system.

[0003] Dendritic cells are highly plastic cells and exist in many subspecies. However, plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) are known to induce type I interferon (IFN), which exhibits growth inhibitory activity against viruses, through activation of Toll-like receptors (TLR).

[0004] pDCs are the main type I IFN-producing cells in the body. Induction of type I IFN production by pDCs requires stimulation of endosomal TLRs, such as TLR7 / TLR9.

[0005] mDCs induce the production of type I IFN through activation of TLR3 and other receptors. In addition, mDCs release inflammatory cytokines, primarily interleukin-12 (IL-12) and tumor necrosis factor-α (TNF-α), upon bacterial infection, inducing the activation of helper T cells (CD4+ T cells).

[0006] However, recent research has revealed the existence of lactic acid bacteria that activate the innate immune system. Patent Document 1 describes that Lactobacillus lactis has the effect of activating pDC and inducing IFN production. Patent Document 2 describes that Lactobacillus helveticus SBT2171 (FERM BP-5445) has the effect of increasing pDC.

[0007] Patent No. 6170190 Patent No. 6705628

[0008] As described in Patent Document 1, Lactobacillus lactis is known to have an IFN production-inducing effect. However, in recent years, with increasing awareness of preventing viral infections such as COVID-19, bacteria that exhibit a more excellent IFN production-inducing effect are desired. Furthermore, SBT2171 described in Patent Document 2 is a bacterial strain selected based on its pDC proliferation-promoting effect, rather than its IFN production-inducing effect. Regarding Lactobacillus helveticus, insufficient research has been conducted in terms of its IFN production-inducing effect.

[0009] In view of the above circumstances, an object of the present invention is to provide a bacterium belonging to Lactobacillus helveticus that has an excellent IFN production-inducing activity.

[0010] As a result of extensive research efforts, the present inventors have discovered a strain of Lactobacillus helveticus that exhibits a more effective type I IFN production induction effect than Lactobacillus lactis JCM5805 specifically described in Patent Document 1 and Lactobacillus helveticus SBT2171 described in Patent Document 2, and have completed the present invention. The present invention, which solves the above-mentioned problems, is as follows.

[0011] [1] A bacterium that can activate plasmacytoid dendritic cells (pDC) and / or myeloid dendritic cells (mDC) and induce type I IFN production, and is classified as Lactobacillus helveticus, which induces 1.5 times or more type I IFN production compared to Lactobacillus helveticus SBT2171 (accession number: Ferm BP-5445).

[0012] [2] Lactobacillus helveticus MCC2430 (Accession number: NITE BP-03882).

[0013] [3] A composition comprising at least one selected from the bacterium according to [1] or [2], a culture of the bacterium, and a treated product of the bacterium.

[0014] [4] The composition according to [3], wherein the composition is a composition for inducing type I IFN production.

[0015] [5] The composition according to [3], wherein the composition is at least one selected from the group consisting of an antiviral composition, an immunostimulating composition, a composition for treating or preventing an immune disease, an anticancer composition, an anti-inflammatory composition, and a composition for preventing a decrease in bone strength.

[0016] [6] The composition according to any one of [3] to [5], which is a food, drink, or pharmaceutical product.

[0017] [7] A method for producing a composition, comprising a step of adding at least one selected from the bacterium according to [1] or [2], a culture of the bacterium, and a treated cell product of the bacterium to a raw material.

[0018] [8] The method according to [7], further comprising a step of fermenting a raw material with the bacterium, wherein the composition is a fermented composition.

[0019] The bacterium of the present invention exhibits an excellent type I IFN production-inducing effect by activating pDC and / or mDC. The composition of the present invention induces type I IFN production in a subject who ingests it, and exhibits antiviral effects, immunostimulating effects, therapeutic or preventive effects for immune diseases, anticancer effects, anti-inflammatory effects, and effects for preventing a decrease in bone strength. The production method of the present invention makes it possible to produce a composition that exhibits the above-mentioned effects.

[0020] 1 is a bar graph showing the concentration of IFNα in the culture supernatant measured by flow cytometry using an IFNα measurement kit. The test was performed three times, and the average value is shown in the graph. The error bars in the graph indicate standard deviation. The p value in the figure was obtained by Student's t-test. Note that for each strain, a 10-fold increase in the amount of sterilized form relative to the dendritic cells was added, but for JCM5805, an additional test was performed in which a 100-fold increase in the amount of sterilized form was added. The label "JCM5805_100" in the figure indicates the test results for JCM5805 in which a 100-fold increase in the amount of sterilized form relative to the dendritic cells was added. This is a bar graph showing the concentration of IFNβ in the culture supernatant measured by ELISA using an IFNβ measurement kit. The test was performed three times, and the average value is shown in the graph. The error bars in the graph indicate standard deviation. The p value in the figure was obtained by Student's t-test. For each strain, a sterilized form was added in an amount 10 times the number of cells compared to the number of dendritic cells, but an additional test was conducted for JCM5805 in which a 100-fold amount of sterilized form was added. The "JCM5805_100" symbol in the figure indicates the test results for JCM5805 in which a 100-fold amount of sterilized form was added compared to the number of dendritic cells.

[0021] Preferred embodiments of the present invention will be described below. However, the present invention is not limited to the following preferred embodiments and can be freely modified within the scope of the present invention. In this specification, percentages are expressed by mass unless otherwise specified.

[0022] <Bacteria capable of inducing type I IFN production> The present invention relates to bacteria classified as Lactobacillus helveticus that can activate plasmacytoid dendritic cells (pDCs) and / or myeloid dendritic cells (mDCs) and induce type I IFN production.

[0023] More specifically, the bacterium of the present invention is a bacterium classified as Lactobacillus helveticus that induces 1.5 times or more type I IFN production compared to Lactobacillus helveticus SBT2171 (accession number: Ferm BP-5445).

[0024] Bacteria classified as Lactobacillus helveticus, which produce 1.5 times or more type I IFN induced by activating pDC and / or mDC compared to SBT2171, are included in the scope of the present invention. The conditions for the test to measure the amount of type I IFN produced to confirm whether or not it belongs to the present invention are not particularly limited. In this specification, for the sake of convenience, the test to confirm whether or not it belongs to the present invention is referred to as a "confirmation test."

[0025] An example of a confirmation test is a test system in which bacterial cells are added to a mixed culture system in which pDCs and / or mDCs are cultured, and the concentration of type I IFN in the culture supernatant is measured. If the measured concentration of type I IFN is 1.5 times or more higher than when SBT2171 bacterial cells are added under the same conditions, it can be determined that the bacterium being the subject of the confirmation test falls within the scope of the present invention.

[0026] The culture system to which the bacterial cells are added may be a culture system in which either pDC or mDC is cultured alone, or may be a mixed culture system of pDC and mDC.

[0027] The scope of the present invention includes any of the following: a form in which pDC is activated to induce the production of type I IFN but mDC is not activated; a form in which mDC is activated to induce the production of type I IFN but pDC is not activated; and a form in which both pDC and mDC are activated to induce the production of type I IFN.

[0028] The bacterial cells added to the culture system of pDC and / or mDC may be live or killed. When killed bacteria are used, they are preferably heat-sterilized.

[0029] There is no limitation on the number of bacterial cells added to a culture system of pDCs and / or mDCs. For example, 10 to 100 times the number of pDCs and / or mDCs can be added. Specifically, a preferred example is a system in which 10 times the number of bacterial cells is added to a culture system of pDCs and / or mDCs.

[0030] The tissue from which the pDCs and / or mDCs used in the confirmation test are derived is not limited. For example, those derived from peripheral blood mononuclear cells or bone marrow cells may be used. The animal species from which they are derived is also not limited, and examples include mammals such as humans, monkeys, mice, rats, rabbits, dogs, and cats. Preferably, pDCs and / or mDCs derived from humans are used.

[0031] The method for measuring the concentration of type I IFN in the culture supernatant of pDC and / or mDC is not particularly limited, and flow cytometry or ELISA can be used. Commercially available kits can also be used for the measurement. Examples include an IFNα measurement kit (Cytometric Bead Array Kit, Becton Dickinson) and an IFNβ measurement kit (Human IFN-beta DuoSet, R&D Systems).

[0032] Mammalian type I IFNs consist of at least eight different classes, including IFNα, IFNβ, IFNδ, IFNε, IFNκ, IFNω, IFNτ, and IFNζ. IFNα, IFNβ, IFNε, IFNκ, and IFNω have been found in humans. The scope of the present invention includes forms that induce the production of any interferon that falls under type I IFN.

[0033] In a preferred embodiment of the present invention, the production of IFNα and / or IFNβ is induced. The scope of the present invention includes an embodiment that induces the production of IFNα but not the production of IFNβ, an embodiment that induces the production of IFNβ but not the production of IFNα, and an embodiment that induces the production of both IFNα and IFNβ. In a preferred embodiment of the present invention, the production of both IFNα and IFNβ is induced.

[0034] The type I IFN measured in the confirmatory test may be any type I IFN interferon, but is preferably IFNα or IFNβ. In this case, the measurement target may be either IFNα or IFNβ, or both IFNα and IFNβ.

[0035] The time from adding the bacterial cells to a pDC and / or mDC culture system until measuring the concentration of type I IFN in the culture supernatant is preferably 12 to 48 hours, preferably 20 to 28 hours, and more preferably 24 hours.

[0036] The confirmatory test may be performed once or multiple times. When the confirmatory test is performed multiple times, the average value of the type I IFN concentration measured in each confirmatory test may be used for comparison with SBT2171. In this case, the test is preferably performed three or more times.

[0037] As described above, the bacterium of the present invention induces type I IFN production that is 1.5-fold or more compared to SBT2171. In a preferred embodiment, the bacterium induces type I IFN production that is 1.8-fold or more, more preferably 2-fold or more, and even more preferably 2.2-fold or more compared to SBT2171.

[0038] In a preferred embodiment, the compound induces 1.5-fold or more IFNα and / or IFNβ production compared to SBT2171. In a preferred embodiment, the compound induces 1.8-fold or more, more preferably 2-fold or more, and even more preferably 2.2-fold or more IFNα and / or IFNβ production compared to SBT2171.

[0039] In a specific embodiment, the production of IFNα is induced at least 1.5 times, preferably at least 1.8 times, more preferably at least 2 times, even more preferably at least 2.2 times, and even more preferably at least 2.3 times, compared to SBT2171.

[0040] In another specific embodiment, the IFNβ production is induced at least 1.5-fold, preferably at least 1.8-fold, more preferably at least 2-fold, even more preferably at least 2.2-fold, even more preferably at least 2.4-fold, even more preferably at least 2.6-fold, even more preferably at least 2.7-fold higher than that of SBT2171.

[0041] In a more specific embodiment, the antibody induces IFNα production that is at least 1.5-fold, preferably at least 1.8-fold, more preferably at least 2-fold, even more preferably at least 2.2-fold, and even more preferably at least 2.3-fold higher than that of SBT2171, and the antibody induces IFNβ production that is at least 1.5-fold, preferably at least 1.8-fold, more preferably at least 2-fold, even more preferably at least 2.2-fold, even more preferably at least 2.4-fold, even more preferably at least 2.6-fold, and even more preferably at least 2.7-fold higher than that of SBT2171.

[0042] The bacteria of the present invention may be in the form of a strain that has been isolated and cloned as a genetically homogeneous bacterial population, or may be in the form of an uncloned, genetically heterogeneous bacterial population.

[0043] Even if the bacterial population is genetically heterogeneous, it is naturally within the scope of the present invention as long as it exhibits the type I IFN production-inducing effect described above as a characteristic of the entire population.

[0044] Among the bacteria of the present invention, an example of a strain that has been isolated and cloned as a genetically homogeneous bacterial population is Lactobacillus helveticus MCC2430 (Accession No.: NITE BP-03882). This strain will be described later.

[0045] The bacterium of the present invention can be easily grown by culturing it. The culturing method is not particularly limited as long as it allows Lactobacillus helveticus to grow, and methods commonly used for culturing lactic acid bacteria can be used with appropriate modifications as necessary. For example, the culture temperature may be 25 to 50°C, preferably 35 to 42°C. Furthermore, the culture is preferably carried out under anaerobic conditions, for example, while aerating with anaerobic gas such as carbon dioxide. Culture may also be carried out under microaerobic conditions, such as liquid static culture.

[0046] The medium for culturing the bacterium of the present invention is not particularly limited, and media commonly used for culturing lactic acid bacteria can be used with appropriate modifications as needed. Specifically, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysates, and blackstrap molasses can be used as carbon sources depending on the assimilation potential. Nitrogen sources include ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, and ammonium nitrate, as well as nitrates. Inorganic salts include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate. Organic components such as peptone, soybean flour, defatted soybean meal, meat extract, and yeast extract may also be used. As a prepared medium, for example, MRS medium can be suitably used.

[0047] The bacterium of the present invention can be provided as a biologically pure culture.

[0048] <Lactobacillus helveticus MCC2430> The present invention also relates to Lactobacillus helveticus MCC2430 (Accession No.: NITE BP-03882), hereinafter simply referred to as MCC2430.

[0049] Based on the following mycological properties and characteristics, MCC2430 of the present invention was deposited as a novel strain at the National Institute of Technology and Evaluation (NPMD) Patent Microorganisms Depositary (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) on April 13, 2023, as Lactobacillus helveticus (Accession Number: NITE BP-03882). This strain is publicly available from the above-mentioned collection institution.

[0050] The MCC2430 of the present invention is not limited to the above-mentioned deposited strain, but may be a strain substantially equivalent to the deposited strain.

[0051] The substantially equivalent strains include strains that fall into at least one of the following (1) to (3): (1) Strains determined to be identical by the Randomly Amplified Polymorphic DNA (RAPD) method or the Pulsed-field gel electrophoresis (PFGE) method (FAO / WHO (2006) Probiotics in Food: Health and Nutritional Properties and Guidelines for Evaluation. Report of a Joint FAO / WHO Expert Consultation on Evaluation of Health and Nutritional Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria, Cordoba, Argentina, 1-4 October 2001 [and] Report of a Joint FAO / WHO Working Group on Drafting Guidelines for the Evaluation of Probiotics in Food, London, Ontario, Canada, 30 April-1 May 2002. FAO Food and Nutrition (Paper 85, Food and Agriculture Organization of the United Nations, World Health Organization, Rome. Page 43) (2) A strain that contains only genes derived from the deposited strain, does not contain any genes of foreign origin, and has a DNA identity of 95% or more. (3) A strain bred from the deposited strain, or a strain that has undergone genetic engineering modification, mutation, or natural mutation, and that has the same level of type I IFN production-inducing activity as the deposited strain when tested by the above-mentioned confirmation test. Furthermore, a strain that has excellent fermentation properties and / or fermentability within the temperature range of normal lactic acid bacteria fermentation is preferred.

[0052] MCC2430 of the present invention can be easily grown by culturing the strain, for example, under the culture conditions and in the medium as described above.

[0053] The MCC2430 of the present invention can be provided as a biologically pure culture.

[0054] <Use of the present bacterium helveticus> The above-described bacterium of the present invention and MCC2430 of the present invention exert a probiotic effect when administered to a subject. For convenience of explanation, the bacterium of the present invention and MCC2430 of the present invention will hereinafter be collectively referred to as the "present bacterium helveticus."

[0055] The present helveticus bacteria can be used in the form of bacteria themselves, a culture, or a processed bacterial cell form. That is, after culturing, the culture obtained may be used as is, or may be diluted or concentrated before use, or bacterial cells recovered from the culture may be used. The present helveticus bacteria may be live or dead, or may contain both live and dead bacteria. Furthermore, the present helveticus bacteria may be a processed bacterial cell form.

[0056] Killed bacteria include those killed by heating, freeze-drying, etc. Other methods for preparing killed bacteria include spray drying, retort sterilization, freeze-drying, UHT sterilization, pressure sterilization, high-pressure steam sterilization, dry heat sterilization, circulating steam disinfection, electromagnetic wave sterilization, electron beam sterilization, high-frequency sterilization, radiation sterilization, ultraviolet sterilization, ethylene oxide gas sterilization, hydrogen peroxide gas plasma sterilization, and chemical sterilization (alcohol sterilization, formalin fixation, electrolyzed water treatment).

[0057] Examples of processed bacterial cells include immobilized bacterial cells immobilized with acrylamide, carrageenan, or the like, cell lysates in which the cell walls and cell membranes of the present Bacillus helveticus cells are partially or completely disrupted, the supernatant obtained by centrifugation thereof (water-soluble fraction), a fraction obtained by partially purifying the supernatant by ammonium sulfate treatment, or a concentrated version of the supernatant. The cell lysate may be obtained by disrupting live or dead cells, or may be obtained by subjecting cells to disruption followed by heating, freeze-drying, or the like.

[0058] Disruption can be performed using methods and equipment known in the art, such as physical disruption, enzyme dissolution treatment, chemical treatment, autolysis treatment, etc. Physical disruption can be performed either in the form of a bacterial cell suspension or in the form of a bacterial cell powder. Examples of physical disruption include disruption by stirring using an ultrasonic homogenizer, homogenizer, ball mill, bead mill, dyno mill, planetary mill, etc.; disruption by pressure using a jet mill, French press, cell disrupter, etc.; and disruption by damaging the bacterial cells through filter filtration. Enzyme dissolution treatment can destroy the cellular structure of the bacterial cells using, for example, enzymes such as lysozyme. Chemical treatment can destroy the cellular structure of the bacterial cells using surfactants such as soybean phospholipids and glycerin fatty acid esters. Autolysis treatment can dissolve the bacterial cells using some of the lactic acid bacteria's own enzymes.

[0059] At least one selected from the present bacterium Helveticus, a culture of the present bacterium Helveticus, and a treated product of the present bacterium Helveticus cells can be used as an active ingredient. Hereinafter, for the sake of convenience, "at least one selected from the present bacterium Helveticus, a culture of the present bacterium Helveticus, and a treated product of the present bacterium Helveticus cells" may be referred to as "the present ingredient."

[0060] The present component may be used for humans or non-human animals (preferably mammals), and is preferably used for humans, pets, and livestock, more preferably for humans. Furthermore, the target population of the present component is not particularly limited as long as it is desired to have a probiotic effect, and examples include infants, children, adults, healthy individuals, middle-aged and elderly individuals, elderly individuals, and individuals with poor intestinal environments. Of these, the present technology is preferably used for infants, adults, and elderly individuals. Furthermore, the present component has few side effects and is highly safe, so it can be taken continuously over a long period of time.

[0061] The subject ingredients may also be useful in treating conditions or diseases that can be prevented, ameliorated or treated by probiotics.

[0062] The present component has the effect of inducing type I IFN production by activating pDCs and / or mDCs in the body of a subject. Therefore, the present component can be used to induce type I IFN production. More specifically, the present component can be used to induce IFNα and / or IFNβ production. Note that this use is not limited to therapeutic use and may be non-therapeutic use. In other words, the present invention also relates to a method for inducing type I IFN production, which comprises administering the present component to a subject. More specifically, the present invention also relates to a method for inducing IFNα and / or IFNβ production, which comprises administering the present component to a subject in need of type I IFN production induction. The present component also relates to a non-therapeutic administration method, which comprises administering the present component to a subject in need of type I IFN production induction. The present component can also be used to manufacture a type I IFN production inducer.

[0063] The subject in need of type I IFN production induction is not particularly limited as long as the subject desires the type I IFN production induction effect, and examples include subjects in need of prevention or treatment of viral infections, subjects in need of immune activation, subjects in need of treatment or prevention of immune diseases, subjects in need of treatment or prevention of cancer, subjects in need of treatment or prevention of inflammation, and subjects in need of prevention of decreased bone strength.

[0064] The type I IFN, whose production is induced by the present component, has the following functions: (1) increasing the viral resistance of cells by suppressing viral replication; (2) increasing the expression of MHC class I molecules in virus-uninfected cells, thereby protecting them from attack by NK cells; and (3) activating NK cells to eliminate virus-infected cells.

[0065] That is, the present component, which has the effect of inducing the production of type I IFN, can be used for the prevention or treatment of viral infections. The use is not limited to therapeutic use, and may be non-therapeutic use. In other words, the present invention also relates to a method for preventing or treating viral infections, which comprises administering the present component to a subject in need of such prevention or treatment. The present component also relates to a non-therapeutic administration method, which comprises administering the present component to a subject in need of such prevention or treatment. The present component can also be used for the manufacture of an agent for the prevention or treatment of viral infections.

[0066] The preventive or therapeutic effect of the present component on viral infections can be confirmed, for example, by a reduction in the incidence of viral infections following ingestion of a composition containing the present component, but the confirmation method is not limited to this.

[0067] The subject in need of prevention or treatment of a viral infection is not particularly limited as long as the subject desires the effect of prevention or treatment of a viral infection, and examples thereof include subjects infected with a virus.

[0068] Type I IFN is involved in the functional expression of macrophages and T cells and also promotes the activation of NK cells, neutrophils, and the like. Type I IFN plays an important role in the innate immune system, and inducing its production can achieve immunostimulatory effects. That is, the present component, which has the effect of inducing type I IFN production, can be used for immunostimulation. Note that this use is not limited to therapeutic use, and may also be non-therapeutic use. In other words, the present invention also relates to an immunostimulatory method comprising administering the present component to a subject in need of immunostimulation. The present invention also relates to a non-therapeutic administration method comprising administering the present component to a subject in need of immunostimulation. The present component can also be used for the manufacture of an immunostimulator.

[0069] The immunostimulatory effect of the present component can be confirmed, for example, by the activation of NK cells and macrophages in peripheral blood upon ingestion of a composition containing the present component, but the confirmation method is not limited to this.

[0070] The subject in need of immunostimulation is not particularly limited as long as the subject desires an immunostimulatory effect, and examples thereof include subjects whose immune cell function has declined due to aging or other reasons.

[0071] Type I IFN has also been recognized as useful in the treatment of autoimmune diseases such as multiple sclerosis, and is being used clinically. Its therapeutic effects on various autoimmune diseases, including colitis, encephalomyelitis, arthritis, and neonatal inflammation, have also been confirmed. That is, the present component, which has the effect of inducing type I IFN production, can be used for the treatment or prevention of immune diseases. This use is not limited to therapeutic use, and may also be non-therapeutic use. In other words, the present invention also relates to a method for treating or preventing immune diseases, which includes administering the present component to a subject in need of such treatment or prevention. It also relates to a non-therapeutic administration method, which includes administering the present component to a subject in need of such treatment or prevention. The present component can also be used for the manufacture of an agent for treating or preventing immune diseases.

[0072] The therapeutic or preventive effect of the present component on immune diseases can be confirmed, for example, by the suppression of T cell activation in peripheral blood by ingestion of a composition containing the present component, but the confirmation method is not limited to this.

[0073] Subjects who require treatment or prevention of immune diseases are not particularly limited as long as they desire the effects of treatment or prevention of immune diseases, and examples thereof include subjects with high levels of autoantibodies in peripheral blood.

[0074] Type I IFN has the effects of inhibiting cell proliferation and activating NK cells, etc., and has been shown to have anti-cancer effects through indirect action via immune responses or direct action on tumor cells, and is also being used clinically. That is, the present component, which has the effect of inducing type I IFN production, can be used for the treatment or prevention of cancer. This use is not limited to therapeutic use, and may also be non-therapeutic use. In other words, the present invention also relates to a method for treating or preventing cancer, which includes administering the present component to a subject in need of cancer treatment or prevention. It also relates to a non-therapeutic administration method, which includes administering the present component to a subject in need of cancer treatment or prevention. The present component can also be used for the manufacture of an agent for treating or preventing cancer.

[0075] The cancer treatment or prevention effect of the present component can be confirmed, for example, by the activation of NK cells in peripheral blood or the extension of progression-free survival (the period from the start of treatment to the progression of cancer or the death of the subject) upon ingestion of a composition containing the present component, but the confirmation method is not limited to these.

[0076] Subjects in need of cancer treatment or prevention are not particularly limited as long as they desire the effects of cancer treatment or prevention, and examples include subjects suffering from renal cell carcinoma.

[0077] Type I IFN mediates the inhibition of proinflammatory gene products through its ability to induce the immunosuppressive cytokine interleukin-10 (IL-10). It also induces other immunosuppressive mediators, such as suppressor of cytokine signaling 1 (SOCS-1) and tristetraprolin (TTP), which act to restore immune system homeostasis through a variety of mechanisms. Thus, the present component, which has the activity of inducing type I IFN production, can be used for the treatment or prevention of inflammation. This use is not limited to therapeutic use, but may also be non-therapeutic use. In other words, the present invention also relates to a method for treating or preventing inflammation, comprising administering the present component to a subject in need of such treatment or prevention. It also relates to a non-therapeutic administration method, comprising administering the present component to a subject in need of such treatment or prevention. The present component can also be used for the manufacture of an anti-inflammatory agent.

[0078] The therapeutic or preventive effect of the present component on inflammation can be confirmed, for example, by an increase in the proportion of regulatory T cells in peripheral blood or the production of IL-10 following ingestion of a composition containing the present component, but the confirmation method is not limited to this.

[0079] Subjects who require treatment or prevention of inflammation are not particularly limited as long as they desire the effects of treating or preventing inflammation, and examples include subjects who are at high risk of chronic inflammation due to aging, obesity, etc.

[0080] Osteoclasts play a key role in bone metabolism, but excessive proliferation leads to a decrease in bone strength. It has been shown that type I IFN inhibits osteoclast formation and contributes to the prevention of bone strength loss. In other words, the present component, which has the effect of inducing type I IFN production, can be used to prevent bone strength loss. The use is not limited to therapeutic use, and may also be non-therapeutic use. In other words, the present invention also relates to a method for preventing bone strength loss, which comprises administering the present component to a subject in need of such prevention. The present invention also relates to a non-therapeutic administration method, which comprises administering the present component to a subject in need of such prevention. The present component can also be used for the manufacture of an agent for preventing bone strength loss.

[0081] The preventive effect of the present component on bone strength loss can be confirmed, for example, by suppressing bone density loss through ingestion of a composition containing the present component, but the confirmation method is not limited to this.

[0082] There are no particular limitations on subjects who need to be prevented from losing bone strength, as long as they desire the preventive effect of losing bone strength, and examples include subjects aged 50 or over who are prone to bone strength loss.

[0083] The administration or ingestion of the present component is preferably for at least one week, more preferably for at least four weeks, and is preferably daily. The amount of the present component used is not particularly limited because it is highly safe, but for example, 1 x 10 ６ ~1 x 10 １２ cfu / kg body weight / day is preferred, and 1 x 10 ７ ~1 x 10 １１ cfu / kg body weight / day is more preferred, and 1 x 10 ８ ~1 x 10 １０ cfu / kg body weight / day is even more preferred.

[0084] The dosage per individual (body weight) is 10 ７ ~10 １４ cfu / day is preferred, 10 ８ ~10 １３ cfu / day is more preferred, 10 ９ ~10 １２cfu / day is more preferred.

[0085] The amount of the component used is preferably 0.01 to 100 mL / kg body weight / day, and more preferably 0.1 to 10 mL / kg body weight / day. In this specification, "cfu" stands for "colony forming unit." When the bacteria are killed, "cfu" can be substituted for "cells."

[0086] <Composition> The present component can be used as is. Alternatively, it can be mixed with a conventional carrier or diluent that is physiologically, pharmaceutically, or food- and beverage-acceptable. That is, the present invention also relates to a composition containing at least one selected from the bacterium of the present invention, a culture of the bacterium, and a processed bacterial cell product of the bacterium, as well as MCC2430 of the present invention, a culture of the strain, and a processed bacterial cell product of the strain. Hereinafter, for convenience of explanation, the composition will be referred to as the composition of the present invention.

[0087] The present components contained in the composition of the present invention exert various pharmacological effects as described above. That is, the composition of the present invention can be provided as a composition for inducing type I IFN production. Furthermore, the composition of the present invention can be provided as at least one type selected from the group consisting of an antiviral composition, an immunostimulating composition, a composition for treating or preventing immune diseases, an anticancer composition, an anti-inflammatory composition, and a composition for preventing a decrease in bone strength.

[0088] The subjects to which the compositions of the present invention are administered are not particularly limited, and include various subjects as described above. That is, the compositions of the present invention can be provided to, for example, those who desire the type I IFN production-inducing effect. Furthermore, the compositions of the present invention can be provided to, for example, subjects who desire a preventive or therapeutic effect against viral infections, subjects who desire an immunostimulating effect, subjects who desire a therapeutic or preventive effect against immune diseases, subjects who desire a therapeutic or preventive effect against cancer, subjects who desire a therapeutic or preventive effect against inflammation, subjects who desire a preventive effect against decreased bone strength, etc.

[0089] The present component can be contained in a composition as an active ingredient, and because it is highly safe, it can be used in a wide range of products, such as pharmaceuticals, foods, and beverages. These products can be produced by using optional components suitable for each application and by known production methods suitable for each application. In this specification, the term "composition" includes food and beverage compositions, pharmaceutical compositions, feed, etc.

[0090] Hereinafter, embodiments of the composition of the present invention when provided as a pharmaceutical product and when provided as a food or drink product will be described in detail.

[0091] <Pharmaceutical Composition> The composition of the present invention can be provided as a pharmaceutical composition. The pharmaceutical composition of the present invention is not particularly limited as long as it contains the present component. The pharmaceutical composition of the present invention may be in a form in which the present component is used as is, or in a form in which the present component is formulated by blending it with a physiologically acceptable liquid or solid pharmaceutical carrier.

[0092] The pharmaceutical composition of the present invention can be safely administered to patients suffering from various diseases. The present invention is expected to cause few side effects even when administered continuously over a long period of time. Furthermore, the present invention can be safely administered to infants and children. Therefore, the present invention is also suitable for preventing, ameliorating, and / or treating diseases or their symptoms in infants and children.

[0093] The present invention may be used for therapeutic or non-therapeutic purposes. "Non-therapeutic purposes" is a concept that does not include medical procedures, i.e., treatment of the human body through therapy. Examples include health promotion and cosmetic procedures. "Improvement" refers to the improvement of a disease, symptom, or condition; the prevention or delay of the worsening of a disease, symptom, or condition; or the reversal, prevention, or delay of the progression of a disease or symptom. "Prevention" refers to the prevention or delay of the onset of a disease or symptom in a subject, or the reduction of the risk of a disease or symptom in a subject.

[0094] The dosage form of the pharmaceutical composition of the present invention is not particularly limited, and specific examples include tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, syrups, suppositories, injections, ointments, patches, eye drops, nasal drops, etc. In addition, additives commonly used as pharmaceutical carriers, such as excipients, binders, disintegrants, lubricants, stabilizers, flavorings, diluents, surfactants, or solvents for injections, can be used in the formulation.

[0095] Furthermore, when formulating, the pharmaceutical composition of the present invention can contain ingredients such as excipients, pH adjusters, colorants, and flavoring agents that are commonly used in formulations. Furthermore, as long as the effects of the present invention are not impaired, the pharmaceutical composition of the present invention can also contain ingredients that have the effect of preventing, ameliorating, and / or treating diseases or symptoms related to the present invention that are known or will be discovered in the future. Additionally, formulation can be carried out by a known method as appropriate depending on the dosage form. When formulating, a pharmaceutical carrier may be added as appropriate.

[0096] The content of the Bacillus helveticus in the pharmaceutical composition of the present invention is appropriately determined depending on the dosage form, dosage method, age and sex of the patient, type of disease, severity of disease, and other conditions, but is usually 1 x 10 ６ ~1 x 10 １２ cfu / g or 1 x 10 ６ ~1 x 10 １２ cfu / mL, preferably in the range of 1 x 10 ７ ~1 x 10 １１ cfu / g or 1 x 10 ７ ~1 x 10 １１ When the present Bacillus helveticus is a killed bacterium, cfu / g or cfu / mL can be replaced with cells / g or cells / mL.

[0097] The administration timing of the pharmaceutical composition of the present invention is not particularly limited, and can be appropriately selected according to the treatment method for the target symptom or disease. It may also be administered prophylactically or for maintenance therapy. The administration form is preferably determined depending on the formulation, the patient's age, sex, other conditions, the severity of the patient's symptoms, etc. In either case, the pharmaceutical composition of the present technology can be administered once a day or multiple times a day, or may be administered once every few days or weeks.

[0098] <Food and drink composition> Furthermore, the composition of the present invention can be used as a food and drink composition. The food and drink composition of the present invention can be produced by adding the present component to a known food or drink, or can be produced as a new food and drink composition by mixing the present component into the raw materials of the food or drink.

[0099] The food and beverage composition of the present invention is not particularly limited as long as it contains the present component. Examples of food and beverage compositions include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit juice drinks, and lactic acid bacteria drinks (including concentrated concentrates and powders for preparation of these beverages); frozen desserts such as ice cream, sherbet, and shaved ice; confectioneries such as candy, chewing gum, candy, gum, chocolate, tablet candy, snacks, biscuits, jelly, jam, cream, and baked goods; dairy products such as processed milk, milk drinks, fermented milk, drinkable yogurt, and butter; bread; enteral nutritional foods, liquid foods such as porridge, baby food, infant formula, sports drinks; and other functional foods. The food and beverage composition may also be a supplement, such as a tablet supplement. In the case of a supplement, the present Helveticus Bacterium can be ingested without being affected by the daily dietary intake and calorie intake of other foods.

[0100] The food and beverage composition of the present invention may contain various ingredients as long as the effects of the present invention are not impaired. For example, the food and beverage composition of the present invention may contain ingredients such as various proteins such as whey protein, casein protein, soybean protein, or pea protein (pea protein), or mixtures or hydrolysates thereof; amino acids such as leucine, valine, isoleucine, or glutamine; carbohydrates such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, dextrin, and starch; human milk oligosaccharides such as 2'-fucosyllactose, 3'-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, lacto-N-tetraose, and lacto-N-neotetraose; vitamins such as vitamin B6 or vitamin C; creatine; citric acid; or fish oil.

[0101] The food and beverage composition of the present invention can be sold as a food and beverage composition or food and beverage labeled for use in inducing type I IFN production, suppressing viral infection, antiviral use, immunostimulation, treating or preventing immune diseases, anticancer use, anti-inflammatory use, or preventing bone strength loss. The food and beverage composition of the present invention can also be sold as a food and beverage composition or food and beverage labeled for use in preventing or treating symptoms or conditions that can be prevented or treated by inducing type I IFN production. The food and beverage composition or food and beverage of the present invention can also be labeled for use in "those who wish to suppress viral infection," "those who wish to maintain immune function," "helps maintain immune function in healthy individuals by acting on plasmacytoid dendritic cells," "those who wish to suppress allergies," "relieves nasal discomfort caused by allergens," "relieves eye discomfort caused by allergens," "those who wish to suppress inflammation," "those who wish to maintain bone components," "those who wish to maintain strong bones," "helps maintain bone health by maintaining bone components," and the like. Needless to say, any other wording describing a secondary effect resulting from the induction of type I IFN production can also be used.

[0102] The food and drink composition of the present invention can be provided and sold as a food and drink composition or food and drink labeled with uses (including health uses) such as probiotics. The food and drink composition or food and drink can be provided and sold with labels indicating that the intended consumers are "those who wish to live with Lactobacillus bacteria," "those who wish to live with lactic acid bacteria," "those who wish to improve their intestinal environment," "those who wish to regulate their stomach condition," "those who wish to create a good intestinal environment," "improve the intestinal environment," etc.

[0103] The term "labeling" as used herein refers to any act intended to inform consumers of the above-mentioned uses. Any labeling that can evoke or infer the above-mentioned uses falls under the category of "labeling" in the present invention, regardless of the purpose, content, object, or medium of labeling. However, labeling in a manner that allows consumers to directly recognize the above-mentioned uses is preferred. Specific examples include the act of describing the above-mentioned uses on the food and beverage composition or food and beverage product or its packaging; the act of transferring, delivering, displaying for transfer or delivery, or importing the product or its packaging on which the above-mentioned uses are described; the act of displaying or distributing the above-mentioned uses in advertisements, price lists, or transaction documents related to the products; or the act of describing the above-mentioned uses in information containing these contents and providing them via electromagnetic means (e.g., the Internet). Labeling on packaging, containers, sales promotion materials such as catalogs, pamphlets, and POP displays, as well as other documents, is particularly preferred.

[0104] Furthermore, the labeling is preferably one approved by the government or the like (e.g., labeling approved under various government systems and made in a manner based on such approval). Examples include labeling as a health food, more specifically, a health food, a functional food, an enteral nutritional food, a special-use food, a nutrient-functional food, a quasi-drug, etc. Other examples include labeling approved by the Consumer Affairs Agency, such as a food for specified health uses, a nutrient-functional food, a food with functional claims, and labeling approved under similar systems. Examples of the latter include labeling as a food for specified health uses, a conditional food for specified health uses, a label indicating an effect on the structure or function of the body, a disease risk reduction label, and a label indicating functionality based on scientific evidence. More specifically, examples include labeling as a food for specified health uses (especially a label indicating health uses) as defined in the Cabinet Office Ordinance on the Permission for Special-Use Labeling under the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009), and similar labeling.

[0105] The content of the present Lactobacillus helveticus in the food and beverage composition of the present invention is appropriately determined depending on the form of the food and beverage composition. ６ ~1 x 10 １２ cfu / g or 1 x 10 ６ ~1 x 10 １２ cfu / mL, preferably in the range of 1 x 10 ７ ~1 x 10 １１ cfu / g or 1 x 10 ７ ~1 x 10 １１ More preferably, it is in the range of cfu / mL.

[0106] The food and beverage composition of the present invention can include powdered infant formula (also known as infant formula). Powdered infant formula includes powdered infant formula for infants aged 0-12 months, follow-up formula for infants aged 6-9 months and older and young children (up to 3 years of age), powdered infant formula for low birth weight infants weighing less than 2500g at birth (low birth weight infants), and various therapeutic formulas used to treat infants with pathological conditions such as cow's milk allergy and lactose intolerance. Furthermore, the composition can be used as a food with health claims or a food for the sick. The health claims system, based on domestic and international trends and consistency with the existing system of foods for specified health uses, covers not only regular foods but also foods in the form of tablets, capsules, etc., and is divided into two categories: foods for specified health uses (individually approved) and foods with nutrient functions (standardized). In addition to powdered milk, infant formula in the present invention can also be in the form of liquid milk.

[0107] <Production Method> A method for producing the composition of the present invention is described below. The method for producing the composition of the present invention includes a step of adding the present component to a raw material. In the present invention, the step of adding the present component may be carried out at any stage in the production process. The method may also include a step of adding the present Lactobacillus helveticus to a raw material and fermenting the raw material. In this case, the composition of the present invention can be provided as a fermented composition.

[0108] The method for producing the composition of the present invention will be exemplified below, but is not limited thereto.

[0109] An example of the production method is a method for producing a composition comprising at least one of the following steps (a) and (b): (a) mixing the present component with a prebiotic; and (b) mixing the present component with a milk component. In this case, a composition containing the present component may be used as the present component. Furthermore, the composition may further contain other probiotics such as Lactobacillus bacteria, lactic acid bacteria, or Bifidobacterium bacteria.

[0110] The step of mixing the prebiotic and / or milk component may occur at any stage of the manufacturing process. For example, the milk component may be mixed at the same time as step (a), or before or after step (a). The milk component is preferably in powder form, and more preferably mixed with the powder of the Lactobacillus helveticus.

[0111] Another example of a method for producing the composition of the present invention includes the following steps (A) and (B): (A) culturing the Bacillus helveticus in a medium to obtain a culture or product containing the Bacillus helveticus; and (B) drying the culture or product to obtain a bacterial powder or a dried product. The drying is preferably performed by freeze-drying or spray-drying.

[0112] The milk components include, but are not limited to, cow's milk, buffalo milk, sheep's milk, goat's milk, horse's milk, skim milk, concentrated skim milk, skim milk powder, concentrated milk, whole milk powder, cream, butter, buttermilk, condensed milk, milk oligosaccharides, and milk proteins, and one or more selected from the group consisting of these can be used. The milk proteins include, but are not limited to, whey, casein, and hydrolysates thereof, and one or more selected from the group consisting of these can be used. Of the milk components, components derived from cow's milk are preferred.

[0113] The milk oligosaccharide is not limited to bovine milk oligosaccharides, but may be, for example, human milk oligosaccharides, including, but not limited to, 2'-fucosyllactose, 3'-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, lacto-N-tetraose, and lacto-N-neotetraose, and one or more of these may be used.

[0114] The hydrolysate can be produced by hydrolyzing the milk component (preferably milk protein), such as whey hydrolysate or casein hydrolysate. Examples of such hydrolysis include acid / alkali hydrolysis and enzymatic hydrolysis. Among these, enzymatic hydrolysis is preferred, and protease is preferred as the enzyme. Examples of such protease include protease, trypepsin, chymotrypsin, plasmin, pepsin, papain, peptidase, and aminopeptidase. The pH during the hydrolysis reaction can be adjusted appropriately to the optimum pH for the enzyme used, for example, at a pH of 2 to 6. The temperature during the hydrolysis reaction is not particularly limited, and is usually preferably in the range of 30 to 60°C, and the reaction time is preferably 2 to 10 hours.

[0115] The form of the present Bacillus helveticus or a culture containing the same is not particularly limited and may be either liquid or solid, but a dried form (e.g., bacterial powder or dried culture) is preferred from the viewpoint of ease of handling during production and storage. Examples of the culture include, as described above, the bacterial cells themselves from which the culture medium has been separated, a culture containing the bacterial cells, and a culture from which the bacterial cells have been removed.

[0116] The drying method is not particularly limited, but examples thereof include spray drying, retort sterilization, freeze drying, UHT sterilization, pressure sterilization, high-pressure steam sterilization, dry heat sterilization, circulating steam disinfection, electromagnetic wave sterilization, electron beam sterilization, high-frequency sterilization, radiation sterilization, ultraviolet sterilization, ethylene oxide gas sterilization, hydrogen peroxide gas plasma sterilization, and chemical sterilization (alcohol sterilization, formalin fixation, electrolyzed water treatment).

[0117] The cells may be disrupted, and the disrupted product may be cells obtained by disrupting live cells or dead cells, or cells that have been disrupted and then subjected to heating, freeze-drying, etc. Of these, from the viewpoint of increasing the viable cell rate, it is preferable to subject the culture to freeze-drying, and from the viewpoint of increasing production efficiency, it is preferable to subject the culture to spray-drying.

[0118] Furthermore, when producing supplements or tablets as the composition of the present invention, the method may include a step of mixing a culture containing the present Lactobacillus helveticus with an excipient or the like to obtain a mixture, and a step of molding the mixture into a predetermined shape. Examples of the molding step include tableting, and an example of tableting is compressing a mixture of powder or granules to obtain a tablet.

[0119] One example of the manufacturing method of the present invention is a method for producing a fermented food or drink (preferably fermented milk) containing the present Bacillus helveticus. The bacterial powder obtained by the above-mentioned manufacturing method is added to a fermented milk raw material, and a step of obtaining fermented milk using the present Bacillus helveticus is carried out. Alternatively, a step of obtaining fermented milk containing the present Bacillus helveticus is carried out by mixing the bacterial powder with fermented milk. The fermentation conditions and blending amounts are adjusted so that the bacterial cell amount of the present Bacillus helveticus in the composition is a predetermined bacterial cell amount. This allows for the provision of a fermented food or drink (preferably fermented milk or a fermented product) containing the present Bacillus helveticus.

[0120] The present invention will be described in more detail below with reference to examples. Note that the examples described below are representative examples of the present invention and should not be construed as narrowing the scope of the present invention.

[0121] (1) Preparation of heat-sterilized lactic acid bacteria strains Four lactic acid bacteria strains listed in Table 1 were statically cultured in MRS medium or M17 medium at 30°C or 37°C for 16 hours. After the culture, the bacteria were collected, washed twice with sterilized water, and then sterilized by autoclaving at 90°C for 15 minutes. Then, 1 x 10 ８ cells / mL or 1 x 10 ９ The concentration was adjusted with sterilized water to 1000 cells / mL to obtain a heat-sterilized sample for use in the test.

[0122]

[0123] (2) Preparation of dendritic cells The dendritic cells (DCs) used in the test were isolated by negative selection from human peripheral blood mononuclear cells (PBMCs) purchased from STEMCELL Technologies, Inc. using the EasySep Human Pan-DC Pre-Enrichment Kit (STEMCELL Technologies, Inc.). The dendritic cells were suspended in RPMI-1640 medium supplemented with antibiotics and heat-inactivated fetal bovine serum (FBS) at a final concentration of 10%, and then cultured at a concentration of 1 x 10. ６ The suspension was adjusted to 1000 cells / mL. The prepared suspension contained a mixture of plasmacytoid dendritic cells (pDCs) and myeloid dendritic cells (mDCs).

[0124] (3) Measurement of type I IFN production by dendritic cells stimulated with bacteria. The dendritic cell suspension prepared in (2) was added with bacteria of various strains prepared in (1), and the cells were incubated with CO ２ Incubated at 37°C and 5% CO ２ The cells were cultured under the following conditions. The sterilized cells of the above four strains were added in an amount 10 times the number of cells relative to the number of dendritic cells. For JCM5805, an additional test was also conducted in which the strain was added in an amount 100 times the number of cells relative to the number of dendritic cells.

[0125] The culture supernatant was collected 24 hours after the start of culture, and the IFNα concentration was measured by flow cytometry using an IFNα measurement kit (Cytometric Bead Array Kit, Becton Dickinson). The IFNβ concentration was measured by ELISA using an IFNβ measurement kit (Human IFN-beta DuoSet, R&D Systems).

[0126] (4) Results The measurement results of the INFα and INFβ production levels of dendritic cells stimulated by the sterilized form of each lactic acid bacteria strain are shown in Figures 1 and 2. Lactobacillus helveticus MCC2430 had the highest ability to induce the production of INFα and INFβ. As described in Japanese Patent No. 6705628 (Patent Document 2), Lactobacillus helveticus SBT2171 is known to contribute to the increase of pDC, and Lactobacillus lactis subsp. lactis JCM5805 is known to activate pDC and promote IFNα production, as described in Japanese Patent No. 6170190 (Patent Document 1). MCC2430 was shown to have significantly higher type I IFN production induction ability than these known strains.

[0127] [Production Examples] Production examples of the pharmaceutical composition and food and drink composition of the present invention are shown below, but the compositions of the present technology are not limited to these.

[0128] [Preparation Example 1] Lactobacillus helveticus MCC2430 is added to MRS liquid medium, and after anaerobic cultivation at 37 ° C for 16 hours, the culture solution is concentrated and freeze-dried to obtain freeze-dried bacterial powder (bacterial powder).The bacterial powder and whey protein concentrate (WPC) are uniformly mixed to obtain a composition.20 g of this composition is dissolved in 200 g of water to obtain a composition containing Lactobacillus helveticus MCC2430 of the present invention.

[0129] [Preparation Example 2] Lactobacillus helveticus MCC2430 is added to MRS liquid medium, and after anaerobic culture at 37 ℃ for 16 hours, the cells are washed three times by centrifugation and resuspended in distilled water, and then suspended in distilled water to 10 mg (cell weight equivalent) / ml, and heat sterilized at 100 ℃ for 15 minutes to obtain a solution containing heat-sterilized cells.The solution containing heat-sterilized cells is concentrated and freeze-dried to obtain the freeze-dried powder (bacterial powder) of the bacteria.The bacterial powder and oligosaccharide are mixed uniformly to obtain a composition. Examples of oligosaccharides that can be used include human milk oligosaccharides such as 2'-fucosyllactose, 3'-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, lacto-N-tetraose, and lacto-N-neotetraose, as well as isomaltooligosaccharides, lactulose, raffinose, fructooligosaccharides, galactooligosaccharides, and soybean oligosaccharides.

[0130] [Production Example 3] Lactobacillus helveticus MCC2430 is added to MRS liquid medium, and after culturing at 37 ° C for 16 hours, the culture solution is concentrated and freeze-dried to obtain freeze-dried bacterial powder (bacterial powder).The bacterial powder and the dried powder of milk protein concentrate (MPC480, manufactured by Fonterra, protein content 80% by mass, casein protein: whey protein = approximately 8:2) are uniformly mixed to obtain a composition.20 g of this composition is dissolved in 200 g of water to obtain a composition containing Lactobacillus helveticus MCC2430 of the present invention.

[0131] [Preparation Example 4] Lactobacillus helveticus MCC2430 is added to MRS liquid medium, and after culturing at 37 ℃ for 16 hours, the culture solution is concentrated and freeze-dried to obtain the freeze-dried powder (bacteria powder) of bacteria containing Lactobacillus helveticus MCC2430 of the present invention.Then, put bacteria powder and crystalline cellulose into agitation granulator and mix.Then, add purified water to granulate, dry the granulated product, obtain the granulated product (pharmaceutical composition) that contains bacterial fermentation components and excipients.In this way, obtain the granulated product that contains Lactobacillus helveticus MCC2430 of the present invention.

[0132] [Production Example 5] The method for producing fermented milk containing Lactobacillus helveticus MCC2430 is described below. First, the raw milk ingredients, water, and other ingredients, if necessary, are mixed, and then preferably homogenized and heat-sterilized. The homogenization and heat-sterilization can be carried out by conventional methods. A lactic acid bacteria starter is added (inoculated) to the heat-sterilized sterilized milk preparation, and the mixture is fermented at a predetermined fermentation temperature to obtain a fermented product. Curd is formed by fermentation. As the lactic acid bacteria starter, for example, lactic acid bacteria commonly used in yogurt production, such as Lactobacillus bulgaricus, Lactococcus lactis, and Streptococcus thermophilus, can be used. When the pH reaches the target value, the formed curd is broken down by stirring and cooled to 10°C or below to obtain a fermented product. Cooling to 10°C or below reduces the activity of lactic acid bacteria, thereby suppressing acid production.

[0133] Next, the fermented product obtained in the fermentation step is heat-treated to obtain a heated fermented product (heat-treated fermented product). By moderately heating the fermented product, acid production by lactic acid bacteria in the heated fermented product can be suppressed. This can prevent a decrease in pH during the subsequent manufacturing process and / or storage. Next, Lactobacillus helveticus MCC2430 is added to the heated fermented product obtained in the heat-treatment step. The amount of Lactobacillus helveticus MCC2430 added is 1 x 10 relative to the heated fermented product. ７ ~1 x 10 １１ cfu / mL is preferred, 1 x 10 ８ ~1 x 10 １０ When Lactobacillus helveticus MCC2430 is a killed bacterium, cfu / mL can be replaced with individual cells / mL.

[0134] After heating, Lactobacillus helveticus MCC2430 is added to the fermented product, followed by concentration. The concentration step can be carried out using any known concentration method. For example, centrifugation or membrane separation can be used. In the centrifugation method, whey is removed from the concentrate to obtain concentrated fermented milk with an increased solid content. In this manner, fermented milk containing Lactobacillus helveticus MCC2430 of the present invention can be produced.

[0135] By ingesting or administering the fermented milk containing Lactobacillus helveticus MCC2430 of the present invention obtained as described above, it is expected that the effects of inducing type I IFN production, suppressing viral infection, antiviral effects, immunostimulating effects, treating or preventing immune diseases, anticancer effects, anti-inflammatory effects, and preventing bone strength loss can be obtained.

[0136] [Production Example 6] The method for producing modified milk powder containing Lactobacillus helveticus MCC2430 is as follows. 10 kg of desalted milk whey protein powder (Mirai Co., Ltd.), 6 kg of milk casein powder (Fonterra Co., Ltd.), 48 kg of lactose (Mirai Co., Ltd.), 920 g of mineral mixture (Tomita Pharmaceutical Co., Ltd.), 32 g of vitamin mixture (Tanabe Pharmaceutical Co., Ltd.), 500 g of lactulose (Morinaga Milk Industry Co., Ltd.), 500 g of raffinose (Nippon Beet Sugar Co., Ltd.), and 900 g of galactooligosaccharide liquid sugar (Yakult Pharmaceutical Co., Ltd.) were dissolved in 300 kg of hot water, and the mixture was further heated and dissolved at 90 ° C for 10 minutes. 28 kg of modified fat (Taiyo Yushi Co., Ltd.) was added and homogenized. The mixture was then sterilized, concentrated, and spray-dried to prepare approximately 95 kg of modified milk powder. To this, Lactobacillus helveticus MCC2430 bacterial powder (1.8 × 10 １１ cfu / g, manufactured by Morinaga Milk Industry Co., Ltd.) was added to prepare approximately 95 kg of Lactobacillus-oligosaccharide formulated powdered milk. The prepared powdered milk was dissolved in water to prepare a formula with a total solids concentration of 14% (w / v), which is the standard formula concentration. The number of Lactobacillus bacteria in the formula was 2.7 x 10 ９cfu / 100 mL can be obtained. The above-mentioned modified milk powder can further contain human milk oligosaccharides such as 2'-fucosyllactose, 3'-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucohexaose I, lacto-N-tetraose, and lacto-N-neotetraose, as well as probiotics such as Lactobacillus bacteria other than Lactobacillus helveticus MCC2430, lactic acid bacteria, and Bifidobacterium bacteria.

[0137] By ingesting or administering the modified milk powder containing Lactobacillus helveticus MCC2430 of the present invention obtained as described above, it is expected that the effects of inducing type I IFN production, suppressing viral infection, antiviral effects, immunostimulating effects, treating or preventing immune diseases, anticancer effects, anti-inflammatory effects, and preventing bone strength loss can be obtained.

[0138] The present invention can be applied to foods, beverages, and pharmaceuticals containing lactic acid bacteria.

Claims

1. A bacterium that can activate plasmacytoid dendritic cells (pDC) and / or myeloid dendritic cells (mDC) and induce type I IFN production, and is classified as Lactobacillus helveticus, which induces 1.5 times or more type I IFN production compared to Lactobacillus helveticus SBT2171 (Accession Number: Ferm BP-5445).

2. Lactobacillus helveticus MCC2430 (Accession Number: NITE BP-03882).

3. A composition comprising at least one selected from the bacterium according to claim 1 or 2, a culture of said bacterium, and a treated product of said bacterium.

4. The composition according to claim 3, wherein the composition is a composition for inducing type I IFN production.

5. The composition according to claim 3, wherein the composition is at least one selected from the group consisting of antiviral compositions, immunostimulating compositions, compositions for treating or preventing immune diseases, anticancer compositions, anti-inflammatory compositions, and compositions for preventing a decrease in bone strength.

6. The composition according to claim 3, which is a food, drink or medicine.

7. A method for producing a composition, comprising the step of adding at least one selected from the bacterium according to claim 1 or 2, a culture of said bacterium, and a treated cell product of said bacterium to a raw material.

8. The method of claim 7, further comprising the step of fermenting a raw material with said bacteria, and said composition is a fermented composition.