Composition for cellular immune activation

Abstract

To provide an IL-12 production promoter containing honey as an active ingredient. [Solution] A composition for activating cellular immunity, containing sunflower honey as an active ingredient.

Description

Technical Field

[0001] The present invention relates to an interleukin-12 (hereinafter also referred to as "IL-12") production promoter, and more specifically, to an IL-12 production promoter containing honey.

Background Art

[0002] Honey is known to have various health and beauty effects in addition to being edible. For example, honey can be used for various purposes as an antibacterial agent (Patent Document 1), a composition for treating wounds, a composition for cosmetics (Patent Documents 2 and 3), and the like.

[0003] IL-12 is a type of cytokine involved in cellular immunity and is secreted from macrophages or dendritic cells, etc. When the concentration of IL-12 in the blood increases, it promotes the differentiation of naive helper T cells into type 1 helper T cells, and activates natural killer cells and cytotoxic T cells, thereby activating cellular immunity.

[0004] Regarding honey and IL-12, Non-Patent Document 1 describes that when orange honey and an antigen are simultaneously administered to rats, the concentration of IL-12 in the blood is significantly higher than when only the antigen is administered.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0006]

Non-Patent Document 1

[0007] The present invention aims to provide an IL-12 production promoter using honey as an active ingredient. [Means for solving the problem]

[0008] The inventors conducted diligent research to achieve the above objective and discovered that culturing conventional dendritic cells in culture media containing honey from different nectar sources promoted IL-12 production in some of the honeys. Furthermore, the inventors found that culturing conventional dendritic cells in culture media containing sunflower honey, linden honey, honeydew honey, or rewarewa honey significantly increased IL-12 production compared to culturing them in culture media containing orange honey, thus completing the present invention.

[0009] In other words, the present invention relates to the following [1] to [7]. [1] An interleukin-12 (IL-12) production promoter comprising at least one honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey as an active ingredient. [2] An interleukin-12 (IL-12) production promoter according to [1], comprising sunflower honey and / or linden honey as an active ingredient. A composition for activating cellular immunity, comprising the IL-12 production promoter described in [3][1] or [2]. A composition for activating cellular immunity that promotes the differentiation of naive helper T cells into type 1 helper T cells, comprising the IL-12 production promoter described in [4][1] or [2]. A composition for activating cellular immunity that activates natural killer cells and / or cytotoxic T cells, comprising an IL-12 production promoter as described in [5][1] or [2]. A composition for the prevention of bacterial and / or viral infections, comprising an IL-12 production promoter as described in [6][1] or [2]. [7] A food composition or pharmaceutical composition, which is an IL-12 production promoter as described in [1] or [2] or a composition as described in any one of [2] to [5].

[0010] Furthermore, the present invention relates to the following [1] to [6]. [1] An interleukin-12 (IL-12) production promoter containing sunflower honey and / or linden honey as active ingredients. A composition for activating cellular immunity, comprising the IL-12 production promoter described in [2][1]. A composition for activating cellular immunity that promotes the differentiation of naive helper T cells into type 1 helper T cells, comprising the IL-12 production promoter described in [3][1]. A composition for activating cellular immunity that activates natural killer cells and / or cytotoxic T cells, comprising the IL-12 production promoter described in [4][1]. A composition for the prevention of bacterial and / or viral infections, comprising the IL-12 production promoter described in [5][1]. [6] A food composition or pharmaceutical composition, which is the IL-12 production promoter described in [1] or any one of [2] to [5]. [Effects of the Invention]

[0011] According to the present invention, by using a specific type of honey, it is possible to provide a highly active IL-12 production promoter with honey as the active ingredient. [Brief explanation of the drawing]

[0012] [Figure 1] Figure 1 is a graph showing the amount of IL-12 produced when conventional dendritic cells were cultured in a medium containing various types of honey in Example 1. [Figure 2]Figure 2 is a graph showing the production amount of IL-12 when normal dendritic cells are cultured in a medium containing various honeys in Example 2.

Embodiments for Carrying Out the Invention

[0013] Hereinafter, embodiments for carrying out the present invention will be described in detail, but the present invention is not limited to the following embodiments.

[0014] One embodiment of the present invention is an interleukin-12 (hereinafter also referred to as "IL-12") production promoter containing at least one kind of honey selected from the group consisting of sunflower honey, bodaiju honey, manna honey, and rewraw honey as an active ingredient.

[0015] Honey is produced mainly from honey collected by bees from plant nectar, sap, secretions of insects parasitic on plants, etc. The type of bees used for honey collection and the production area of honey are not particularly limited. In the present embodiment, honey is at least one kind selected from the group consisting of sunflower honey, bodaiju honey, manna honey, and rewraw honey, and may be sunflower honey and / or bodaiju honey, may be manna honey and / or bodaiju honey, may be used alone, or two, three, or four kinds may be used in combination.

[0016] Sunflower honey is honey that can be collected by bees from the flowers of sunflowers (Helianthus annuus) belonging to the genus Helianthus of the Asteraceae family.

[0017] Bodaiju honey is honey that can be collected by bees from the flowers of bodaiju (Tilia miqueliana) belonging to the genus Tilia of the Malvaceae family.

[0018] Honeydew honey, known as "forest honey" in Europe, is made from the sap of various trees (such as oak, pine, fir, and spruce) that insects ingest. After the insects have absorbed the sap, they leave only the sugars in the form of droplets on the leaves and trunks of the trees, which bees then collect and process into honey. Honeydew honey contains more minerals and amino acids than typical flower-derived honey and, like manuka honey, is known to have high antibacterial properties.

[0019] Rewarewa honey is honey collected from the flowers of Knightia excelsa, a plant belonging to the Knightia genus of the Proteaceae family. Like Manuka honey and honeydew honey, it is known to have high antibacterial properties.

[0020] Honey can be obtained, for example, as a beekeeping product in accordance with conventional laws.

[0021] While stored in the beehive, honey is generally kept at around 35°C by the bees' body temperature. After being harvested from the beehive, honey is generally heated as needed to reduce its viscosity, facilitate transportation and packaging, and melt any crystals.

[0022] The heating temperature for honey may be 59°C or lower, 58°C or lower, 57°C or lower, 56°C or lower, 55°C or lower, 54°C or lower, 53°C or lower, 52°C or lower, 51°C or lower, 50°C or lower, 49°C or lower, 48°C or lower, 47°C or lower, 46°C or lower, 45°C or lower, 44°C or lower, 43°C or lower, 42°C or lower, 41°C or lower, 40°C or lower, 39°C or lower, 38°C or lower, 37°C or lower, or 36°C or lower. The heating temperature for honey may be, for example, 35°C or higher, 36°C or higher, or 37°C or higher. A heating temperature of 35°C or higher is preferable in terms of handling the honey, as it reduces the viscosity of the honey and makes it easier to reduce crystallization in the honey.

[0023] Honey after harvesting may be used as is, or it may be purified by removing proteins, waxes, debris, bee and hive residue, filtering, extraction with organic solvents, fractionation, sterilization, concentration, etc., before or after heating.

[0024] Honey is composed of approximately 80% carbohydrates, with the remainder consisting of other healthy components such as vitamins, minerals, flavonoids, organic acids, and fatty acids, and the remainder being water. The majority of the carbohydrates in honey are glucose and fructose, with small amounts of oligosaccharides and maltose also present.

[0025] In this embodiment, the honey content in the IL-12 production promoter is the total amount of at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, if sunflower honey and linden honey are used in combination, the total amount of sunflower honey and linden honey), for example, 0.1% by mass or more, 1% by mass or more, 3% by mass or more, 5% by mass or more, 7% by mass or more, and 10% by mass or more. , 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 70% by mass % or more, 75% by mass or more, 80% by mass or more, 85% by mass or more, 90% by mass or more, 95% by mass or more, 97% by mass or more, 98% by mass or more, 99% by mass or more, 99.5% by mass or more, or 100% by mass. The content of honey in the composition is, for example, 99.5% by mass or less, 99% by mass or less, 98% by mass or less, 97% by mass or less, 95% by mass or less, 90% by mass or less, 85% by mass or less, 80% by mass or less, 75% by mass or less, 70% by mass or less, 65% by mass or less, 60% by mass or less, 55% by mass or less % or less, 50% by mass or less, 45% by mass or less, 40% by mass or less, 35% by mass or less, 30% by mass or less, 25% by mass or less, 20% by mass or less, 15% by mass or less, 10% by mass or less, 7% by mass or less, 5% by mass or less, 3% by mass or less, 2% by mass or less, or 1% by mass or less.

[0026] An IL-12 production promoter according to one embodiment may further contain, in addition to honey, other known components that promote IL-12 production. Examples of known components that promote IL-12 production include lactic acid bacteria such as Lactobacillus acidophilus, bifidobacteria such as Bifidobacterium breve, and yeasts such as Saccharomyces cerevisiae.

[0027] The ability to promote IL-12 production can be evaluated by using conventional dendritic cells (cDCs), obtained by differentiating bone marrow cells, adding the test substance, and quantifying the amount of IL-12 in the culture medium after a certain period of incubation using ELISA. If the amount of IL-12 produced increases compared to when honey is added (negative control), it can be evaluated as having an IL-12 production promoting effect. Furthermore, the strength of the IL-12 production promoting effect can be evaluated by the rate of increase compared to the amount of IL-12 produced in the negative control.

[0028] The IL-12 production promoter according to one embodiment may consist solely of at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, sunflower honey and / or linden honey), or it may be a composition containing at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, sunflower honey and / or linden honey) and other components. Furthermore, the IL-12 production promoter according to one embodiment may be a food composition or a pharmaceutical composition. Food compositions and pharmaceutical compositions will be described below.

[0029] Another aspect of the present invention is a composition for activating cellular immunity comprising an IL-12 production promoter according to one embodiment. IL-12 is a type of cytokine involved in cellular immunity, and when the concentration of IL-12 in the blood increases, cellular immunity is activated by promoting the differentiation of naive helper T cells into type 1 helper T cells. Furthermore, when the concentration of IL-12 in the blood increases, cellular immunity is activated through the activation of natural killer cells and / or cytotoxic T cells. Therefore, the composition for activating cellular immunity according to one embodiment can promote the differentiation of naive helper T cells into type 1 helper T cells, or activate natural killer cells and / or cytotoxic T cells.

[0030] Another aspect of the present invention is a composition for preventing bacterial and / or viral infections, comprising an IL-12 production promoter according to one embodiment. In cellular immunity, bacteria are phagocytosed by macrophages and the like, which are activated by cytokines produced by type 1 helper T cells. Cells infected with viruses are killed by natural killer cells or cytotoxic T cells and the like. Therefore, a composition comprising an IL-12 production promoter according to one embodiment can activate the body's defense mechanisms against bacteria and / or viruses by promoting the differentiation of naive helper T cells into type 1 helper T cells and activating natural killer cells and / or cytotoxic T cells, and can therefore be used as a composition for preventing bacterial and / or viral infections.

[0031] A composition for activating cellular immunity according to one embodiment may consist only of at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, sunflower honey and / or linden honey), or it may be a composition containing at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, sunflower honey and / or linden honey) and other components. A composition for activating cellular immunity according to one embodiment may be a food composition or a pharmaceutical composition.

[0032] The honey content in the above composition is the total amount of at least one type of honey selected from the group consisting of sunflower honey, linden honey, honeydew honey, and rewarewa honey (for example, if sunflower honey and linden honey are used in combination, the total amount of sunflower honey and linden honey), for example, 0.1% by mass or more, 1% by mass or more, 3% by mass or more, 5% by mass or more, 7% by mass or more, 10% by mass or more, 15% by mass or more. Above, 20% by mass or more, 25% by mass or more, 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, 70% by mass or more , 75% by mass or more, 80% by mass or more, 85% by mass or more, 90% by mass or more, 95% by mass or more, 97% by mass or more, 98% by mass or more, 99% by mass or more, 99.5% by mass or more, or 100% by mass. The content of honey in the composition is, for example, 99.5% by mass or less, 99% by mass or less, 98% by mass or less, 97% by mass or less, 95% by mass or less, 90% by mass or less, 85% by mass or less, 80% by mass or less, 75% by mass or less, 70% by mass or less, 65% by mass or less, 60% by mass or less, 55% by mass or less % or less, 50% by mass or less, 45% by mass or less, 40% by mass or less, 35% by mass or less, 30% by mass or less, 25% by mass or less, 20% by mass or less, 15% by mass or less, 10% by mass or less, 7% by mass or less, 5% by mass or less, 3% by mass or less, 2% by mass or less, or 1% by mass or less.

[0033] The composition according to the present invention may be a food composition, a pharmaceutical composition (pharmaceutical), or a quasi-drug, and may be a food composition or a pharmaceutical composition. Furthermore, honey may be used as a food composition (food) as is, or it may be used as an additive to food.

[0034] It is preferable that the food composition (food) emphasizes the tertiary function of the food (physical condition regulation function). Examples of foods that emphasize the tertiary function of the food include health foods, functional foods, nutritional compositions, nutritional supplements, supplements, health foods, foods for specified health uses, nutrient function foods, or foods with functional claims.

[0035] When used as an additive to food, the food may contain ingredients that are acceptable as food. Examples of ingredients that are acceptable as food include minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, cooling agents, binders, sweeteners, disintegrants, lubricants, colorants, flavorings, stabilizers, preservatives, sustained-release regulators, surfactants, solvents, and humectants.

[0036] If the food composition is a nutritional supplement or similar product, it may be in any form, such as solid, liquid, or paste, and may also be in dosage forms such as tablets (including uncoated tablets, sugar-coated tablets, effervescent tablets, film-coated tablets, chewable tablets, lozenges, etc.), capsules, pills, powders, granules, liquids, suspensions, emulsions, syrups, or pastes. These can be prepared, for example, by mixing honey with other ingredients as needed.

[0037] Food compositions may be obtained by adding honey to food, and examples of food include the following, and by mixing an effective amount of honey into intermediate or final products during the manufacturing process of these, food for the above purpose can be obtained: beverages such as soft drinks such as coffee, juice and tea beverages, milk beverages, lactic acid bacteria beverages, yogurt beverages, carbonated beverages, and alcoholic beverages such as sake, Western liquor, fruit wine and honey wine; spreads such as custard cream; pastes such as fruit paste; Western confectionery such as chocolate, donuts, pies, cream puffs, gum, jelly, candy, cookies, cakes and puddings; Japanese confectionery such as daifuku, mochi, manju, castella, anmitsu and yokan; frozen desserts such as ice cream, ice pops and sherbet; cooked foods such as curry, gyudon, zosui, miso soup, soup, meat sauce, pasta, pickles and jam; and seasonings such as dressings, furikake, umami seasonings and soup bases.

[0038] The method for producing the food composition is not particularly limited and can be carried out according to known methods as appropriate. For example, the food composition can be obtained for the above purpose by mixing the honey with an intermediate or final product in the manufacturing process of the food composition.

[0039] Pharmaceuticals or quasi-drugs may be in any form, such as solid, liquid, or paste, and may be in dosage forms such as tablets (including uncoated tablets, sugar-coated tablets, effervescent tablets, film-coated tablets, chewable tablets, lozenges, etc.), capsules, pills, powders, granules, liquids, suspensions, emulsions, syrups, pastes, or injections (including cases where the product is prepared as a liquid by mixing it with distilled water or an infusion solution such as an amino acid solution or electrolyte solution at the time of use). These various preparations can be prepared, for example, by mixing the above-mentioned honey, which is the active ingredient, with other ingredients as needed and molding them into the above-mentioned dosage forms.

[0040] In addition to honey, pharmaceutical compositions may include pharmaceutically acceptable ingredients, such as pharmaceutically acceptable additives (excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents, etc.) and food-grade ingredients (for example, minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, cooling agents, binders, sweeteners, disintegrants, lubricants, colorants, flavorings, stabilizers, preservatives, sustained-release regulators, surfactants, solvents, and wetting agents).

[0041] The daily intake of honey contained in the composition is not particularly limited as long as it has an IL-12 production-promoting effect, and may be, for example, 0.01 to 70 g / day, with 4 to 70 g / day being more preferable. The composition according to this embodiment may be taken once a day, or it may be taken in multiple doses, such as twice or three times a day. Furthermore, the timing of intake is not particularly limited and may be before meals, after meals, between meals, or before going to bed. [Examples]

[0042] The present invention will be described more specifically below based on examples. However, the present invention is not limited to the following examples.

[0043] [Example 1: Investigation of the interleukin-12 production promoting effect of various types of honey (1)] Bone marrow cells (4.0 × 10) collected from normal C57BL / 6J mice 6 Cells (Cells / mL) were cultured for 7 days in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) to induce differentiation into conventional dendritic cells (cDCs). To the culture medium, orange honey, sunflower honey, or linden honey diluted with sterile water to a final concentration of 1%, or lipopolysaccharide (LPS) diluted with sterile water to a final concentration of 1 μg / mL as a positive control, was added (IL-12 production induction step). After further incubation for 48 hours, the amount of IL-12 in the culture medium was quantified by ELISA. ELISA MAX was used for quantification. TM Standard Set Mouse IL-12 / IL-23 (p40) was used. Figure 1 shows the results, with the amount of IL-12 obtained represented as 100% for the group to which LPS was added and 0% for the group to which the IL-12 production induction process was not performed.

[0044] As a result, in all groups to which honey was added, an increase in IL-12 production was observed compared to the group that did not undergo the IL-12 production induction process. Furthermore, the amount of IL-12 produced when sunflower honey (LPS ratio 68.7%) or linden honey (LPS ratio 60.4%) was added was significantly higher than when orange honey (LPS ratio 34.5%) was added, indicating 2.0 times and 1.7 times higher IL-12 production activity, respectively.

[0045] [Example 2: Investigation of the interleukin-12 production promoting effect of various types of honey (2)] Differentiation into normal dendritic cells was induced in the same manner as in Example 1. To the culture medium, orange honey, honeydew honey, or rewarewa honey diluted with sterile water to a final concentration of 1%, or lipopolysaccharide (LPS) diluted with sterile water to a final concentration of 1 μg / mL as a positive control, was added (IL-12 production induction step). After further incubation for 48 hours, the amount of IL-12 in the culture medium was quantified by ELISA in the same manner as in Example 1. The results of the obtained IL-12 amounts are shown in Figure 2, with the amount of IL-12 in the group to which LPS was added set to 100% and the amount of IL-12 in the group that did not undergo the IL-12 production induction step set to 0%.

[0046] As a result, in all groups to which honey was added, an increase in IL-12 production was observed compared to the group that did not undergo the IL-12 production induction process. Furthermore, the amount of IL-12 produced when honeydew honey (LPS ratio 71.8%) or rewarewa honey (LPS ratio 110.5%) was added was significantly higher than when orange honey (LPS ratio 27.9%) was added, indicating 2.6 times and 4.0 times higher IL-12 production activity, respectively (n=6).

Claims

1. A composition for activating cellular immunity, containing sunflower honey as an active ingredient.

2. A cellular immune activation composition according to claim 1, for promoting the differentiation of naive helper T cells into type 1 helper T cells.

3. The cellular immune activation composition according to claim 1, for activating natural killer cells and / or cytotoxic T cells.

4. A composition for activating cellular immunity according to any one of claims 1 to 3, based on its IL-12 production-promoting effect.

5. A food composition or a pharmaceutical composition, which is a cellular immune activation composition according to any one of claims 1 to 3.

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