Composition for activating dendritic cells

Abstract

The objective of this invention is to provide a dendritic cell activation composition and a dendritic cell activation method that are highly safe and can be easily consumed. [Solution] The dendritic cell activation composition according to the present invention contains an extracellular polysaccharide produced from the Streptococcus thermophilus JCM17834T strain. According to the present invention, a dendritic cell activation composition that is safer and easier to ingest than conventional methods can be provided. Furthermore, by administering this dendritic cell activation composition to a target that requires it, the dendritic cells of that target can be activated.

Description

[Technical Field]

[0001] This invention relates to a composition for activating dendritic cells. [Background technology]

[0002] To date, drugs effective against diseases such as cancer, AIDS, and herpesvirus have been proposed (see, for example, Patent Documents 1, 2, and 3). Some of these drugs, when used in combination with dendritic cell activators in patients who need them, improve dendritic cell activity, thereby treating or alleviating the symptoms of the diseases listed above. Furthermore, it has been found that there is a close relationship between "dendritic cell activation" and "maintaining or improving health." Therefore, developing drugs that can activate dendritic cells is becoming increasingly important in today's society where health awareness is on the rise.

[0003] However, taking such drugs solely for the purpose of activating dendritic cells can have some side effects. Therefore, for example, in humans, they may not always be safe, especially for infants and the elderly. In other words, the target population for taking such drugs is limited by the age and health condition of the individual. Therefore, there is a need for a dendritic cell activating composition that has fewer side effects than the aforementioned drugs and can be easily taken by a wide range of people regardless of age or other factors. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Special Publication No. 2003-502008 [Patent Document 2] Special Publication No. 2015-513401 [Patent Document 3] Special Publication No. 2021-531765 [Overview of the project] [Problems that the invention aims to solve]

[0005] The objective of this invention is to provide a dendritic cell activation composition and a dendritic cell activation method that are highly safe and can be easily consumed. [Means for solving the problem]

[0006] To solve the above problems, the inventors conducted diligent research and found that by administering a composition containing extracellular polysaccharides produced from Streptococcus thermophilus JCM17834T to subjects such as humans, it is possible to activate the dendritic cells of those subjects, particularly plasmacytoid dendritic cells, thus completing the present invention.

[0007] In other words, the present invention is as follows:

[0008] (1) A dendritic cell activation composition containing extracellular polysaccharides produced from Streptococcus thermophilus JCM17834T.

[0009] (2) The dendritic cell activating composition according to (1), wherein the dendritic cells are plasmacytoid dendritic cells.

[0010] (3) The dendritic cell activation composition according to (2), wherein the dendritic cell activation is an increase in the CD86 expression level of the plasmacytoid dendritic cells in peripheral blood mononuclear cells.

[0011] (4) The dendritic cell activation composition according to (2) or (3), wherein the dendritic cell activation is an increase in the CD40 expression level of the plasmacytoid dendritic cells in peripheral blood mononuclear cells.

[0012] (5) The dendritic cell activation composition according to (2) or (3), wherein the dendritic cell activation is an increase in the HLA-DR expression level of the plasmacytoid dendritic cells in peripheral blood mononuclear cells.

[0013] (6) The composition for activating dendritic cells according to (2), wherein said dendritic cell activation is an increase in the CD40 expression level and HLA-DR expression level of said plasmacytoid dendritic cells in peripheral blood mononuclear cells.

[0014] (7) The composition for activating dendritic cells according to (2), wherein said dendritic cell activation is an increase in the CD86 expression level, CD40 expression level and HLA-DR expression level of said plasmacytoid dendritic cells in peripheral blood mononuclear cells.

[0015] (8) A method for activating dendritic cells, which uses an exopolysaccharide produced by Streptococcus thermophilus JCM17834T.

[0016] (9) The method for activating dendritic cells according to (8), wherein said dendritic cells are plasmacytoid dendritic cells.

[0017] The present invention also includes the following inventions.

[0018] (10) A method for activating dendritic cells in a subject, which comprises administering a composition containing an exopolysaccharide produced by Streptococcus thermophilus JCM17834T to a subject in need thereof.

[0019] (11) Use of an exopolysaccharide produced by Streptococcus thermophilus JCM17834T for activating dendritic cells.

[0020] (12) An exopolysaccharide produced by Streptococcus thermophilus JCM17834T, which is used as an active ingredient of a composition for activating dendritic cells.

[0021] (13) Use of an exopolysaccharide produced by Streptococcus thermophilus JCM17834T for the manufacture of a composition that activates dendritic cells.

[0022] (14) An exopolysaccharide produced by Streptococcus thermophilus JCM17834T, which activates dendritic cells. [Effects of the Invention]

[0023] According to the present invention, a composition for activating dendritic cells that is safer and can be ingested more easily than conventional methods can be provided. Further, dendritic cells in a subject can be activated by allowing the subject in need of the composition for activating dendritic cells to ingest the composition. Furthermore, by using an extracellular polysaccharide produced from *Streptococcus thermophilus* JCM17834T, a safer and simpler method for activating dendritic cells can be provided. [Brief Description of Drawings]

[0024] [Figure 1] It is a graph showing the expression intensity of CD86 in pDC in the JCM17834T-derived EPS non-addition group and the JCM17834T-derived EPS addition group in Example 1. In the figure, "non-addition group" refers to the "JCM17834T-derived EPS non-addition group", and "JCM17834T" refers to the "JCM17834T-derived EPS addition group". [Figure 2] It is a graph showing the expression intensity of HLA-DR in pDC in the JCM17834T-derived EPS non-addition group and the JCM17834T-derived EPS addition group in Example 1. In the figure, "non-addition group" refers to the "JCM17834T-derived EPS non-addition group", and "JCM17834T" refers to the "JCM17834T-derived EPS addition group". [Figure 3] It is a graph showing the expression intensity of CD40 in pDC in the JCM17834T-derived EPS non-addition group and the JCM17834T-derived EPS addition group in Example 1. In the figure, "non-addition group" refers to the "JCM17834T-derived EPS non-addition group", and "JCM17834T" refers to the "JCM17834T-derived EPS addition group". [Mode for Carrying Out the Invention]

[0025] The composition according to the embodiment of the present invention contains extracellular polysaccharides produced from bacteria belonging to the Streptococcus thermophilus, and has the function of activating dendritic cells, particularly plasmacytoid dendritic cells. It is preferable that this composition be fermented milk. The definitions of each term in the present invention are described in detail below.

[0026] 1. Streptococcus thermophilus The bacteria belonging to Streptococcus thermophilus according to the embodiments of the present invention are a general term for all bacteria that have been taxonomically recognized as belonging to the genus Streptococcus thermophilus. Preferably, the bacteria belonging to Streptococcus thermophilus are bacteria belonging to the genus Streptococcus that have the ability to produce extracellular polysaccharides. Although bacteria belonging to Streptococcus thermophilus may be classified as either plant-derived or animal-derived depending on their origin, in the present invention, either plant-derived or animal-derived lactic acid bacteria can be used. Preferably, the bacterial strain of Streptococcus thermophilus in the present invention is one or more bacterial strains belonging to Streptococcus thermophilus that can prepare fermented milk (e.g., yogurt) with a sufficient history of safe consumption. Such bacteria belonging to Streptococcus thermophilus are preferably those that have a 16S rRNA gene with 90% or more sequence identity with the nucleotide sequence of Sequence ID No. 1 below (i.e., all or characteristic parts of the 16S rRNA gene sequence (V1 region, V2 region, V3 region, or all or part of the V1, V2, and V3 regions, or part including the V1, V2, and V3 regions, etc.)), more preferably those that have a 16S rRNA gene with 95% or more sequence identity with the nucleotide sequence of Sequence ID No. 1, even more preferably those that have a 16S rRNA gene with 98% or more sequence identity with the nucleotide sequence of Sequence ID No. 1, and 16S rRNA gene with 99% or more sequence identity with the nucleotide sequence of Sequence ID No. 1. It is even more preferable that the bacterium is a member of Streptococcus thermophilus that possesses an rRNA gene, and it is particularly preferable that the bacterium is a member of Streptococcus thermophilus that possesses a 16S rRNA gene having 99.5% or more sequence identity with the base sequence of Sequence ID No. 1.The sequence number 1 shown in the following sequence listing is the sequence of the V1-V3 region of the 16S rRNA gene of Streptococcus thermophilus JCM17834T (corresponding to the nucleotide sequence from position 1 to 588 in the full-length sequence of the 16S rRNA gene) (hereinafter sometimes referred to as the "nucleotide sequence").

[0027] (Sequence Listing) <16S rRNA gene (SEQ ID NO: 1)> CCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCGGCACTGAATCCCGGAAAGGATCCAACACCTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTT CGCTCCCACGCTTTCGAGCCTCAGCGTCAGTTACAGACCAGAGAGCCGCTTTCGCCACCGGTGTTCCTCCATATATCTACGCATTTCACCGCTACACATGGAATTCCACTCTCCCCTTCTGCACTCAAGTTTGACAGTTTCCAAAG CGAACTATGGTTGAGCCACAGCCTTTAACTTCAGACTTATCAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTCGGGACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTCCCTTTCTGGTAAGCTA CCGTCACAGTGTGAACTTTCCACTCTCACACCCGTTCTTGACTTACAACAGAGCTTTACGATCCGAAAACCTTCTTCACTCACGCGGCGTTGCTCGGTCAGGGTTGCCCCCATTGCCGAAGATTCCCTACTGCTGCCTCCCGTAGGA

[0028] In embodiments of the present invention, sequence identity refers, unless otherwise specified, to the ratio of matching bases shared between two sequences when they are optimally aligned. Analysis of base sequence identity can be performed using algorithms or programs well known to those skilled in the art (e.g., BLASTN, BLASTP, BLASTX, ClustalW). When using a program, parameters can be appropriately set by those skilled in the art, or the default parameters of each program may be used. Specific methods for these analysis methods are also well known to those skilled in the art. Commercially available genetic information processing software may be used for calculating identity.

[0029] Furthermore, the bacterium belonging to Streptococcus thermophilus according to the embodiment of the present invention is preferably Streptococcus thermophilus JCM17834T (hereinafter sometimes referred to as "JCM17834T").

[0030] In the present invention, the "JCM17834T strain" may include strains equivalent to the JCM17834T strain, as long as the effects of the present invention are obtained. Here, equivalent strains refer to derivative strains, mutant strains, or progeny strains of these derivative or mutant strains of the JCM17834T strain, and which have the same mycological properties as JCM17834T. Preferably, the "JCM17834T strain" in the present invention is a strain that has the same mycological properties as JCM17834T and also has the ability to activate dendritic cells.

[0031] Furthermore, "Streptococcus thermophilus JCM17834T" can be obtained from the RIKEN BioResource Center, Microbial Materials Development Laboratory (3-1-1 Takanodai, Tsukuba City, Ibaraki Prefecture) under accession number: JCM17834T (see, for example, "https: / / www.jcm.riken.jp / cgi-bin / jcm / jcm_number?JCM=17834").

[0032] 2. Exopolysaccharide The extracellular polysaccharide (hereinafter sometimes referred to as "EPS") according to the embodiment of the present invention is produced from bacteria belonging to the Streptococcus thermophilus mentioned above. Preferably, the extracellular polysaccharide according to the embodiment of the present invention is produced from Streptococcus thermophilus JCM17834T.

[0033] Incidentally, the extracellular polysaccharides according to the embodiments of the present invention are preferably polysaccharides produced from the Streptococcus thermophilus JCM17834T strain, its derivatives, mutants, or progeny strains of these derivatives or mutants, and more preferably the constituent sugars are partially or entirely the same polysaccharides as those of JCM17834T.

[0034] The extracellular polysaccharides referred to herein are not particularly limited as long as they have the desired effect, but are preferably those produced by fermenting raw milk (preferably raw materials containing milk as defined in the Order Concerning Standards for Ingredients of Milk and Dairy Products) with the lactic acid bacteria described above. The extracellular polysaccharides used in the embodiments of the present invention may be one type or a combination of two or more types. Furthermore, extracellular polysaccharides produced by fermenting plants themselves (such as seeds) or plant-derived raw materials with the lactic acid bacteria described above may be excluded. Furthermore, extracellular polysaccharides produced by fermenting culture media or substrates that do not contain one or more of glucose, galactose, or lactose with the lactic acid bacteria described above may be excluded.

[0035] 3. Dendritic cells Dendritic cells (sometimes referred to as "DCs") are a type of antigen-presenting cell that plays a central role in the body's immune response. They are widely distributed in tissues that come into contact with the outside world, such as the skin and mucous membranes, as well as in the lymphatic system, including lymph nodes and the spleen. One of the main types of DCs is the plasmacytoid dendritic cell.

[0036] Plasmacytoid dendritic cells (sometimes referred to as "pDCs") are a type of dendritic cell found in peripheral blood mononuclear cells (sometimes referred to as "PBMCs") and play an important role in the immune system. pDCs, in particular, contribute to protection against viral infection and the regulation of the immune response. pDCs have the ability to produce large amounts of type I interferons such as IFN-α and IFN-β, and this interferon production promotes the suppression of viral replication and the activation of immune cells.

[0037] The compositions according to embodiments of the present invention activate pDCs, as described in the examples below. Here, pDC activation refers to a change in the state in which pDCs enhance the immune response, and specifically refers to an increase in the expression intensity (e.g., Geometric Mean fluorescence intensity, gMFI) of cell surface markers such as CD86, HLA-DR, and CD40. For this reason, it is known that checking the expression intensity of cell surface markers such as CD86, HLA-DR, and CD40 is a method to confirm the degree of pDC activation. Here, an increase in the expression intensity of cell surface markers such as CD86, HLA-DR, and CD40 means that the average expression level of each cell surface marker such as CD86, HLA-DR, and CD40 is significantly higher compared to the baseline value without the addition of a specific stimulus (e.g., the addition of EPS produced from bacteria belonging to Streptococcus thermophilus). Furthermore, an increase in the average expression level of cell surface markers such as CD86, HLA-DR, and CD40 means that the expression amount of the cell surface marker has increased due to the specific stimulus. The degree of pDC activation according to the embodiment of the present invention is confirmed by checking the expression intensity of CD86, HLA-DR, CD40, etc., based on the method described in the examples below.

[0038] CD86 is a co-stimulatory molecule that indicates the activated state of pDCs and promotes T cell activation. When pDCs are activated by the composition according to the embodiment of the present invention, the expression level of CD86 increases. In this case, the CD86 expression level is preferably 1.1 times, more preferably 1.3 times, and even more preferably 1.4 times compared to the baseline value without any specific stimulus.

[0039] HLA-DR is an antigen-presenting molecule that indicates the activation state of pDCs, and CD4 + An immune response is induced through antigen presentation to T cells. When pDCs are activated by the composition according to the embodiment of the present invention, the expression level of HLA-DR increases. In this case, the expression level of HLA-DR is preferably 1.5 times, more preferably 1.6 times, even more preferably 2 times, particularly preferably 2.5 times, and most preferably 3 times, compared to the baseline value without any specific stimulation.

[0040] CD40 is a co-stimulatory molecule that indicates the activated state of pDCs and modulates the immune response through interaction with T cells and B cells. When pDCs are activated by the composition according to the embodiment of the present invention, the expression level of CD40 increases. In this case, the CD40 expression level is preferably 1.3 times, more preferably 1.4 times, even more preferably 2 times, and particularly preferably 2.5 times compared to the baseline value without any specific stimulus.

[0041] 4. Composition The term "composition" as used herein includes preparations such as liquid foods, supplements, and food additives, food and beverages (excluding plants and animals themselves), and food and beverage compositions (including processed food and beverages), and other substances that can be ingested by animals (including humans). Food and beverages may also include fermented milk (yogurt). Furthermore, "composition" may also refer to pharmaceuticals or quasi-drugs.

[0042] As described above, the compositions according to the embodiments of the present invention contain extracellular polysaccharides produced from bacteria belonging to the Streptococcus thermophilus. Extracellular polysaccharides are a type of metabolite produced by the culture and fermentation of these lactic acid bacteria. For example, Streptococcus thermophilus JCM17834T is known to produce extracellular polysaccharides during its culture, and the compositions according to the embodiments of the present invention contain these extracellular polysaccharides as active ingredients. In the compositions according to the embodiments of the present invention, the lower limit of the daily intake of these extracellular polysaccharides is 10 μg, preferably 15 μg, more preferably 25 μg, even more preferably 100 μg, even more preferably 200 μg, particularly preferably 500 μg, even more preferably 1.0 mg, even more preferably 2.0 mg, and even more preferably 3.0 mg. Furthermore, the upper limit is not particularly limited, but for example, it is 200 mg, preferably 100 mg, more preferably 70 mg, even more preferably 30 mg, even more preferably 8.0 mg, particularly preferably 7.0 mg, and most particularly preferably 5.0 mg. In the composition according to the embodiment of the present invention, the range of the daily intake of the extracellular polysaccharide can be freely set by combining the above-mentioned upper and lower limits.

[0043] In the production of the composition according to the embodiment of the present invention, the stage of incorporating the extracellular polysaccharide can be appropriately selected. The stage of incorporation is not particularly limited as long as the properties of the extracellular polysaccharide are not significantly impaired. For example, a culture containing the extracellular polysaccharide obtained by culturing bacteria belonging to Streptococcus thermophilus that produce extracellular polysaccharides, or a crude or purified product thereof, can be mixed with the raw materials and incorporated.

[0044] The compositions according to embodiments of the present invention can be in the form of single-serving unit packaging, and can also be in the form of containing effective bacterial cells per unit package.

[0045] Furthermore, for example, it is preferable that the extracellular polysaccharide be included in each unit package so that it is ingested in the range of 1 μg / g to 1000 μg / g, more preferably in the range of 10 μg / g to 800 μg / g, even more preferably in the range of 50 μg / g to 500 μg / g, even more preferably in the range of 100 μg / g to 500 μg / g, and even more preferably in the range of 200 μg / g to 400 μg / g, and 250 μg / g to 350 μg / g. Note that these upper and lower limits can be freely combined.

[0046] When packaging a composition according to an embodiment of the present invention per unit package, known packaging can be used. For example, there are no particular restrictions on the type of packaging, such as paper, plastic, glass, nylon, stainless steel, aluminum, iron, copper, silver, or bamboo. However, considering that lactic acid bacteria are facultative anaerobic bacteria, it is preferable to use a form that does not come into contact with air or oxygen. For example, it is preferable to include a step in the manufacturing process or packaging process of the composition according to an embodiment of the present invention to eliminate the possibility of contact with oxygen, and it is preferable to select a packaging material that does not allow oxygen to permeate into the packaging during storage after packaging.

[0047] The composition according to the embodiment of the present invention is not particularly limited in terms of the route of intake, but is preferably taken orally. Furthermore, the form in which the composition is taken orally is not particularly limited, but is particularly preferred to be taken orally as "fermented milk".

[0048] Furthermore, the form of consumption of the compositions according to the embodiments of the present invention is not limited to the "fermented milk" described above, but may also be, for example, chocolate, jelly, ice cream, frozen dessert, beverage, chewable tablets, tablets, gummies, cheese, spread, health foods (nutrient functional foods, foods for specified health uses, nutritional supplements, functional foods, supplements, etc.), pharmaceuticals, quasi-drugs, etc. In the present invention, "functional foods" includes health foods to which health claims based on the food standards of Codex Alimentarius (FAO / WHO Joint Food Standards Committee) apply. In the manufacture of these compositions, the stage at which the active ingredient, the bacterial cells, are incorporated can be appropriately selected. In addition, the laws of each country in which the food is implemented shall take precedence regarding the display of functional foods.

[0049] 7. Fermented milk As described above, the compositions according to the embodiments of the present invention are preferably consumed as "fermented milk." Fermented milk according to the embodiments of the present invention refers to milk (animal milk) that has been fermented, and includes, but is not limited to, "fermented milk," "lactic acid bacteria beverage," "milk beverage," and "natural cheese" as defined in the Ministerial Ordinance on Standards for Ingredients of Milk and Dairy Products (Milk and Dairy Products Ordinance). For example, fermented milk refers to, but is not limited to, "fermented milk" as defined in the Milk and Dairy Products Ordinance, that is, milk such as raw milk, cow's milk, special milk, raw goat's milk, pasteurized goat's milk, raw sheep's milk, adjusted milk, low-fat milk, non-fat milk, and processed milk, or milk containing an equivalent or greater amount of non-fat milk solids, which is fermented with lactic acid bacteria or yeast and made into a solid (hard type), paste (soft type), or liquid (drink type), or frozen, but is not limited to these.

[0050] Furthermore, commercially available products containing the bacteria described in the embodiments of the present invention may be used for convenience.

[0051] In the fermented milk according to the embodiment of the present invention, the concentration range of non-fat milk solids is preferably, for example, within the range of 4.0% by mass or more and 12.0% by mass or less, more preferably within the range of 6.0% by mass or more and 10.0% by mass or less, and still more preferably within the range of 7.0% by mass or more and 9.0% by mass or less. Further, the concentration of milk fat is preferably, for example, within the range of 0.2% by mass or more and 4.0% by mass or less, more preferably within the range of 0.3% by mass or more and 3.0% by mass or less, and still more preferably within the range of 0.4% by mass or more and 2.0% by mass or less.

[0052] In the fermented milk according to the embodiment of the present invention, the exopolysaccharide produced by bacteria belonging to Streptococcus thermophilus per unit package is 10 6 CFU / g or more and 10 12 CFU / g or less, it is preferable to contain the exopolysaccharide so as to be ingested within the range, 10 6 CFU / g or more and 10 11 X CFU / g or less, it is more preferable to contain the exopolysaccharide so as to be ingested within the range, 10 7 CFU / g or more and 10 10 CFU / g or less, it is more preferable to contain the exopolysaccharide so as to be ingested within the range, 10 8 CFU / g or more and 10 9 CFU / g or less, it is more preferable to contain the exopolysaccharide so as to be ingested within the range. Note that the upper limit values and lower limit values can be freely combined.

[0053] A typical example of fermented milk is yogurt. In the present invention, the term "yogurt" encompasses the yogurt defined by the FAO / WHO. Such yogurt includes, for example, plain yogurt, hard yogurt (set-type yogurt), soft yogurt, drink yogurt and the like. As the yogurt according to the embodiment of the present invention, hard yogurt is particularly preferable from the viewpoints of satiety and feeling of fullness. Further, when the subject is an elderly person, hard yogurt is particularly preferable also from the viewpoint of preventing aspiration.

[0054] An example of a method for preparing fermented milk according to an embodiment of the present invention is "a method in which raw milk is sterilized and cooled, a lactic acid bacteria starter used by those skilled in the art is added to the raw milk, and the raw milk containing the lactic acid bacteria starter is fermented at a fermentation temperature and for a fermentation time such that a predetermined lactic acid acidity is achieved." In this method, the lactic acid acidity is preferably in the range of 0.6 to 1.2, and more preferably in the range of 0.6 to 0.8. The fermentation temperature is preferably in the range of 40°C to 45°C. The fermentation time is preferably in the range of 2 hours to 12 hours, and more preferably in the range of 3 hours to 8 hours.

[0055] In the fermented milk according to the embodiment of the present invention, fermentation microorganisms such as lactic acid bacteria other than those belonging to Streptococcus thermophilus, bifidobacteria, and yeast may be added as a lactic acid bacteria starter or for the purpose of imparting functionality, or they may not be included.

[0056] The fermented milk according to the embodiment of the present invention can be easily consumed. Furthermore, fermented milk has a long history of safe consumption and is free from concerns about side effects, so consumers can consume it with peace of mind.

[0057] By packaging the fermented milk according to the embodiment of the present invention in an appropriate amount for a single serving, it becomes possible to consume the fermented milk according to the embodiment of the present invention appropriately and easily, which is preferable from the standpoint of usability. Although the appropriate amount for a single serving varies from person to person, in the case of fermented milk with a non-fat milk solids content of 8.0% by mass, it is preferable that the amount per serving is in the range of 10 mL to 1000 mL, more preferably in the range of 30 mL to 500 mL, even more preferably in the range of 50 mL to 200 mL, and preferably 80 mL to 120 mL. Alternatively, it is preferable that the amount per serving is in the range of 10 g to 1000 g, more preferably in the range of 30 g to 500 g, more preferably in the range of 50 g to 200 g, more preferably in the range of 80 g to 150 g, and even more preferably in the range of 100 g to 120 g.

[0058] In embodiments of the present invention, the "individual packaging form" encompasses all forms, such as common packaging forms including containers with lids, bottles with caps, individual bags, pouches, and tubes. In embodiments of the present invention, the uses of the fermented milk according to the embodiments of the present invention can be clearly indicated by including a description of the uses, effects, and methods of consumption of the fermented milk according to the embodiments of the present invention in each individual package, or in a package including multiple individual packages, and / or by enclosing a document containing such a description, and / or by displaying a separate pamphlet or other document containing such a description.

[0059] The compositions according to the embodiments of the present invention are preferably ingested continuously for 3 days or more, preferably for 5 days or more, preferably for 1 week or more, preferably for 2 weeks or more, preferably for 4 weeks or more, more preferably for 6 weeks or more, even more preferably for 8 weeks or more, even more preferably for 10 weeks or more, even more preferably for 12 weeks or more, even more preferably for 24 weeks or more, and particularly preferably for 36 weeks or more, from the viewpoint of enhancing the dendritic cell activation effect. The compositions according to the embodiments of the present invention have a sufficient history of safe consumption and can be ingested safely, so there is no particular upper limit on the duration of intake, and it can be continued indefinitely. However, if an upper limit were to be set, it would be, for example, 60 weeks or less. This upper limit could also be, for example, 120 weeks or less, 100 weeks or less, or 80 weeks or less. Oral ingestion is preferred as the mode of intake.

[0060] When packaging a composition according to an embodiment of the present invention on a unit-package basis, known packaging materials can be used. For example, there are no particular limitations, such as paper, plastic, glass, nylon, stainless steel, aluminum, iron, copper, silver, or bamboo. However, considering that lactic acid bacteria are facultative anaerobic bacteria, it is preferable to use a form that does not come into contact with air or oxygen. For example, it is preferable to include steps in the manufacturing process and packaging process of the composition according to the embodiment of the present invention to eliminate the possibility of contact with oxygen, and it is preferable to select a packaging material that does not allow oxygen to permeate into the packaging during storage after packaging.

[0061] The composition according to the embodiment of the present invention is not particularly limited in terms of the route of intake, but is preferably taken orally. Furthermore, the form in which the composition is taken orally is not particularly limited, but is particularly preferred to be taken orally as "fermented milk".

[0062] Furthermore, the form of intake of the composition according to the embodiment of the present invention is not limited to the "fermented milk" described above, but can take the form of food and beverage compositions, pharmaceuticals, therapeutic foods, supplements, food additives, and other formulations.

[0063] In embodiments of the present invention, the formulation is an oral preparation prepared by conventional methods using additives that are permissible for formulation. This formulation can take the form of solid preparations such as tablets, powders, granules, capsules, pills, and sustained-release preparations, or liquid preparations such as solutions, suspensions, and emulsions. Examples of additives that are permissible for formulation include excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, flavorings, buffers, antioxidants, and pH adjusters. Specifically, food additives include seasonings such as processed seasonings, flavorings, and cooking mixes.

[0064] Furthermore, in embodiments of the present invention, food and beverages and food and beverage compositions are processed for consumption by humans or animals and are not particularly limited to any form that can be taken orally, such as a solution, suspension, emulsion, powder, or solid molded product. Examples of food and beverages and food and beverage compositions include, specifically, dairy products such as milk beverages (including processed milk), yogurts, lactic acid bacteria beverages, fermented milk, ice creams, creams, and cheeses; powdered beverages such as soft drinks, fruit juices, vegetable drinks, soy milk beverages, coffee beverages, tea beverages, jelly drinks, cocoa, smoothies, sports powders, nutritionally fortified powders, beauty powders, powdered soups, steamed bun mixes, concentrated beverages, and alcoholic beverages; wheat flour products such as bread, pasta, noodles, cake mixes, fried chicken batter, and breadcrumbs; chocolate, gum, candy, This includes confectionery such as cookies, gummies, snacks, Japanese sweets, jellies, puddings and other desserts; retort foods such as curry, pasta sauce, pot-au-feu, stew, and Japanese-style dishes; oils and fats such as processed oils, butter, margarine, spreads, and mayonnaise; instant foods such as freeze-dried foods; processed agricultural products such as canned agricultural products, jams and marmalades, pickles, boiled beans, cereals, and rice porridge; processed marine products; processed livestock products; frozen foods such as pizza, doria, gratin, prepared dishes, and fried foods; liquid foods, and even animal feed, tablets, and cosmetics for oral use.

[0065] Furthermore, in embodiments of the present invention, examples of food and beverages and food and beverage compositions include alcoholic beverages such as whiskey, bourbon, spirits, liqueurs, wine, fruit wine, sake, Chinese liquor, shochu, beer, non-alcoholic beer with an alcohol content of 1% or less, sparkling sake, other miscellaneous alcoholic beverages, and chuhai; processed foods such as processed egg products, processed seafood and meat products (including offal such as liver) (including delicacies), soups such as miso soup, seasonings such as miso, soy sauce, furikake, and other seasoning condiments, and liquid foods such as concentrated liquid foods. Mineral water includes both sparkling and non-sparkling mineral water.

[0066] In the embodiments of the present invention, the foods and beverages and food and beverage compositions include classifications such as functional foods, health nutrition foods, health foods, foods for specified health uses, foods with functional claims, nutrient function foods, foods for the sick, infant formula, infant formula for pregnant or lactating women, or foods and beverages with disease risk reduction claims. Here, disease risk reduction claims refer to claims for foods and beverages that have the potential to reduce disease risk, and are claims established or approved based on or referencing the standards set by the FAO / WHO Joint Food Standards Committee (Codex Alimentarius Commission). When making claims regarding the functionality of food, the laws of each country regarding functional foods and food safety must be followed (for example, the system for health foods, the system for functional health foods, the Dietary Supplements Health Education Act, Structure / Function Claim, European Food Safety Authority guidance, etc.). Furthermore, when making claims regarding the functionality of food, the claims must be made after being approved by the laws of each country regarding functional foods and food safety. Furthermore, when making claims regarding the functionality of food products, the claims must be made in accordance with the functional food laws and food safety laws of each country.

[0067] Incidentally, it is preferable to display a description of the composition according to the embodiment of the present invention, such as its use, efficacy, function, type of active ingredient, and method of use. The term "display" as used herein includes all displays that inform consumers of the above-mentioned effects. This display only needs to be one that can evoke or infer the above-mentioned content, and may include all displays regardless of the purpose of the display, the content of the display, or the object or medium on which it is displayed. Specifically, the term "display" as used herein includes displaying the above-mentioned explanation on the product packaging or container, displaying or distributing the above-mentioned explanation on advertisements, price lists or transaction documents related to the product, providing information containing these by electromagnetic means (such as the internet), and sales talk. The term "display" as used herein also includes displays that recommend the consumption of the composition according to the embodiment of the present invention for a specific target, so-called "notifications (including notifications from meal management apps, nutrition management apps, healthcare apps, etc.)."

[0068] Furthermore, it is preferable that products packaged with a composition according to an embodiment of the present invention be labeled with phrases such as, for example, "activates dendritic cells," "assists (supports) dendritic cells," "maintains dendritic cell activity," "reduces the risk of decreased dendritic cell activity," "acts on dendritic cells," "easily activates dendritic cells," "easily maintains dendritic cell activity," "for dendritic cell activation," "for maintaining dendritic cell activity," "supports the maintenance of dendritic cell activity," "helps enhance dendritic cell activity," "activates dendritic cells, which are the command center of the immune system," and "maintains or improves people's health by acting on dendritic cells." It is also possible to indicate a period such as "temporary" or "long-term" at the beginning of each phrase as appropriate.

[0069] Furthermore, it is preferable that products packaged with a composition according to an embodiment of the present invention be labeled with statements such as, for example, "activates plasmacytoid dendritic cells," "assists (supports) plasmacytoid dendritic cells," "maintains the activity of plasmacytoid dendritic cells," "reduces the risk of decreased plasmacytoid dendritic cell activity," "acts on plasmacytoid dendritic cells," "easily activates plasmacytoid dendritic cells," "easily maintains the activity of plasmacytoid dendritic cells," "for activating plasmacytoid dendritic cells," "for maintaining the activity of plasmacytoid dendritic cells," "supports the maintenance of plasmacytoid dendritic cell activity," "helps enhance the activity of plasmacytoid dendritic cells," "activates plasmacytoid dendritic cells, which are the command center of the immune system," and "maintains or improves people's health by acting on plasmacytoid dendritic cells." It is also possible to appropriately indicate a period such as "temporary" or "long-term" at the beginning of each statement.

[0070] Furthermore, the wording used to display such information is not limited to the examples given above; any wording that has a similar meaning is acceptable.

[0071] In embodiments of the present invention, the composition may contain other ingestible components, various additives, pharmaceutical raw materials, etc., as components other than lactic acid bacteria.

[0072] The above-mentioned composition can also be made into special-use foods, complete nutritional foods, nutritional supplements, foods for specified health uses, foods with functional claims, processed foods, etc. Furthermore, the composition can be incorporated into beverages other than drinkable yogurt, liquid foods, etc. The composition can also be used after drying, concentration, or other processing.

[0073] The compositions or fermented milk according to embodiments of the present invention are suitable for ingestion or administration to subjects for whom dendritic cell activation is desirable. There are no particular limitations on the subjects referred to herein, but if we were to give specific examples, they would include subjects who wish to maintain their health, subjects who are aware of having a weak constitution, subjects who are aware of nonspecific symptoms, and subjects who wish to obtain the effect of dendritic cell activation (including healthy individuals and sick individuals).

[0074] The present invention will be described in more detail below based on examples. These examples are not intended to limit the present invention. [Examples]

[0075] (Evaluation study of pDC activity of EPS derived from Streptococcus thermophilus JCM17834T using human peripheral blood mononuclear cells) (1) Preparation of EPS derived from Streptococcus thermophilus JCM17834T EPS derived from JCM17834T was purified from a culture obtained by culturing JCM17834T in a 10% skim milk powder medium supplemented with 0.5% yeast extract. Specifically, cells to which JCM17834T was added were cultured at 37°C for 18 hours under normal pressure, either with or without the addition of bacteria. Trichloroacetic acid was added to the culture to a final concentration of 10% by mass to remove denatured proteins and obtain a protein-depleted culture. Then, three times the volume of cold ethanol was added to the obtained protein-depleted culture supernatant. The protein-depleted culture with cold ethanol added was then allowed to stand overnight at 4°C under normal pressure to obtain a first precipitate containing EPS derived from JCM17834T. Then, the solution containing the first precipitate, obtained by dissolving this first precipitate in MilliQ water, was dialyzed using a dialysis membrane (molecular weight cutoff 6000-8000 Da). After dialyzing, nucleic acids and proteins in the solution containing the first precipitate were enzymatically degraded using "DNase (Sigma-Aldrich K.K.)", "RNase (Sigma-Aldrich K.K.)", and "Proteinase K (Sigma-Aldrich K.K.)". Subsequently, the solution containing the first precipitate after enzymatic degradation was subjected to ethanol precipitation again using three times the volume of ethanol of the supernatant to obtain a second precipitate. Then, the solution containing this second precipitate, obtained by dissolving it in MilliQ water, was dialyzed again using a dialysis membrane (molecular weight cutoff 6000-8000 Da). The solution containing the second precipitate after dialyzing was freeze-dried to purify the EPS derived from JCM17834T.

[0076] (2) Evaluation of pDC activity using EPS derived from JCM17834T PBMC (Precision for Medicine) is placed in a 96-well microwell plate (Iwaki) in a 5x10 ratio. 5Cells were seeded at a rate of cells / well. Subsequently, JCM17834T-derived EPS was added to each well of a 96-well microwell plate seeded with PBMCs, so that the final concentration of JCM17834T-derived EPS was 200 μg / mL. The cells in each well were then cultured at 37°C and 5% CO2 for 24 hours. As a Negative Control, EPS purification was performed from a medium without JCM17834T, and the purified product was added to the same well plate so that the final concentration was 200 μg / mL. The cells in each well were then cultured at atmospheric pressure at 37°C and 5% CO2 for 24 hours, as described above. After that, each culture was collected, and the solution containing each culture was centrifuged at 1300 rpm for 3 minutes at 4°C. The supernatant obtained by centrifugation was removed, and the mixture was washed twice with PBS (FACS buf.) containing 2% FCS. Human BD Fc Block® (manufactured by BD) was added to the remaining precipitate and allowed to stand at room temperature for 10 minutes. Subsequently, the following cell-staining dyes were added to the precipitate: FVD780 (eBiosciense), Anti-Human Lineage Cocktail 1 (CD3, CD14, CD16, CD19, CD20, CD56) FITC (BD), Anti-HLA-DR APC (BD), Anti-CD11c BV421 (BD), Anti-CD123 BV510 (BD), Anti-CD304 PE (BD), Anti-CD86 PE-Cy7 (registered trademark) (BD), and Anti-CD40 PerCP-Cy5.5 (registered trademark) (Biolegend). The samples were incubated at 4°C for 30 minutes in the dark to stain the cells in the precipitate and obtain evaluation samples. These evaluation samples were then washed twice with FACS buf. Subsequently, Fixation and Permeabilization Solution (BD) was added to the washed evaluation sample and incubated at 4°C for 20 minutes in the dark to fix the stained cells in the evaluation sample. Then, the evaluation sample was washed twice with 1× Permeabilization buffer (BD), suspended in FACS buffer, and measured using BD FACSLyric (BD).Cells that were "Lineage Cocktail 1 negative, HLA-DR positive, CD11c negative, CD123 positive, and CD304 positive" were defined as "pDCs," and the expression intensities (Geometric Mean fluorescence intensity, gMFI) of CD86, HLA-DR, and CD40 were analyzed. PBMCs purchased from Precision for Medicine were used, and the evaluation was performed three times (n=3). The culture medium used was RPMI 1640 (Gibco, Thermo Fisher SCIENTIFIC) supplemented with 10% FCS (BioWest) and 1% Penicillin-Streptomycin (Gibco, Thermo Fisher SCIENTIFIC).

[0077] In the group treated with JCM17834T-derived EPS, the expression levels of the pDC activation markers CD86, HLA-DR, and CD40 were significantly higher in all PBMC lots compared to the group without EPS (see Figures 1 to 3). Therefore, it was revealed that JCM17834T-derived EPS increases the expression levels of CD86, HLA-DR, and CD40 in pDCs. [Industrial applicability]

[0078] The composition according to the present invention can activate DCs, particularly pDCs. Therefore, ingesting the composition according to the present invention can contribute to maintaining or improving the health of humans and other organisms. [Accession Number]

[0079] JCM17834T

Claims

1. Contains extracellular polysaccharides produced by Streptococcus thermophilus strain JCM17834T. A composition for activating dendritic cells.

2. The dendritic cells are plasmacytoid dendritic cells. The dendritic cell activation composition according to claim 1.

3. The aforementioned dendritic cell activation is an increase in CD86 expression in the plasmacytoid dendritic cells in peripheral blood mononuclear cells. The dendritic cell activation composition according to claim 2.

4. The aforementioned dendritic cell activation is an increase in CD40 expression levels of the plasmacytoid dendritic cells in peripheral blood mononuclear cells. The dendritic cell activation composition according to claim 2.

5. The aforementioned dendritic cell activation is an increase in HLA-DR expression levels of the plasmacytoid dendritic cells in peripheral blood mononuclear cells. The dendritic cell activation composition according to claim 2.

6. The aforementioned dendritic cell activation is an increase in the CD40 expression level and HLA-DR expression level of the plasmacytoid dendritic cells in peripheral blood mononuclear cells. The dendritic cell activation composition according to claim 2.

7. The aforementioned dendritic cell activation is an increase in the expression levels of CD86, CD40, and HLA-DR in the plasmacytoid dendritic cells in peripheral blood mononuclear cells. The dendritic cell activation composition according to claim 2.

8. A method for activating dendritic cells using extracellular polysaccharides produced from the Streptococcus thermophilus JCM17834T strain.

9. The dendritic cells are plasmacytoid dendritic cells. The method for activating dendritic cells according to claim 8.

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