immunostimulant

Abstract

Provided is a new immunostimulant.　The features of the present invention for solving the problem are as follows. 1. An agent for promoting production of IL-6 in dendritic cells, the agent comprising, as an active ingredient, at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0]. 2. The agent for promoting production of IL-6 according to 1. described above, the agent being characterized in that the production of IL-6 is carried out through binding to TLR2 and / or TLR4. 3. An agent for causing proliferation of helper T-cells by dendritic cells, the agent comprising, as an active ingredient, at least one selected from GlcCer[d18:2(4E,8Z) / 18:0] and GlcCer[d18:2(4E,8Z) / 26:0]. 4. An activator for presenting an antigen to dendritic cells, the activator comprising, as an active ingredient, GlcCer[d18:2(4E,8Z) / 18:0]. 5. An immunostimulant comprising, as an active ingredient, at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0].

Description

immunostimulants

[0001] The present invention relates to an immunostimulant, which is widely used in medicines, health foods, foods, etc.

[0002] When pathogens or bacteria enter the body through cuts or mucous membranes, dendritic cells, which are distributed throughout the body, recognize and phagocytose the antigens, and present them to white blood cells and lymphocytes. Neutrophils, macrophages, and natural killer cells then phagocytose and kill the pathogens that have invaded the body (innate immunity). These cells are used to evaluate immunostimulatory effects.

[0003] There are two types of defense responses against foreign substances: innate immunity and adaptive immunity. In the innate immune response, immune cells such as neutrophils, dendritic cells, and macrophages produce cytokines in response to innate immune activators derived from bacteria or viruses, which then triggers an immune response. The innate immune system is an infection defense mechanism shared by all living organisms, and is generally nonspecific, allowing for a rapid response and effective function against many sources of infection.

[0004] Innate immune responses in dendritic cells and other cells are enhanced when pathogens or antigens bind to pattern recognition receptors (PRRs), activating NF-kB and other cytokines, resulting in the release of various cytokines, such as IL-6 and TNF-α. PRRs include Toll-like receptors (TLRs) and C-type lectin receptors (CLRs: Dectin-1, DC-SIGN, Mincle (Macrophage-Inducible C-type Lectin), etc.), each of which binds to different types of antigens (Figure 2). Dendritic cells and macrophages recognize antigens via PRRs and present them to neutrophils and T cells (antigen presentation). Once activated, the presented cells recognize the presented antigen and phagocytose and kill it, activating the immune response. Furthermore, activation signals for each cell after antigen presentation are mediated by costimulatory molecules. For example, CD40, 80, 86, and MHC I and II are expressed on dendritic cells, and CD80 and 86 bind to CD28 on T cells, while CD40 binds to CD40L molecules on helper T cells, thereby activating each other.

[0005] Dendritic cells also have phagocytic ability and release IL-6. When dendritic cells mature, they have superior antigen-presenting ability compared to other cells and release cytokines. Various cytokines are released, including IL (interleukin)-6, which is produced in response to LPS stimulation. Furthermore, mature dendritic cells activate naive T cells into effector T cells and activate other immune cells. For example, they promote the migration of neutrophils toward pathogens and the phagocytic ability of macrophages. Furthermore, mature dendritic cells induce the differentiation of naive T cells into Th17 cells by IL-6 and other factors, promoting neutrophil migration.

[0006] Against this background, the present inventors discovered that novel sphingolipids promote IL-6 production in dendritic cells, thereby enhancing innate immunity, and thus completed the present invention. Furthermore, by testing the type of PRR to which the novel sphingolipids bind, they found that they bind to TLR2 and 4, thereby mediating antigen presentation, thereby completing the present invention. Furthermore, by performing a mixed lymphocyte reaction (MLR) test in which dendritic cells were co-cultured with human peripheral blood mononuclear cells (PBMCs) to evaluate the proliferation ability of CD4+ helper T cells, they found that the novel sphingolipids enhance the proliferation ability of helper T cells and enhance antigen presentation ability, thereby completing the present invention. Furthermore, by evaluating cell membrane surface markers (CD40, 80, 86) related to antigen presentation ability, they found that the novel sphingolipids increase the expression of CD40 and 80 on dendritic cells, thereby completing the present invention. Thus, the present invention aims to provide a novel immunostimulant that activates innate immunity.

[0007] The features of the present invention for solving the above problems are as follows: 1. An agent for promoting IL-6 production in dendritic cells, which comprises at least one active ingredient selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0]. 2. The agent for promoting IL-6 production according to 1. above, wherein IL-6 production is mediated by binding to TLR2 and / or TLR4. 3. An agent for expanding helper T cells by dendritic cells, which comprises at least one active ingredient selected from GlcCer[d18:2(4E,8Z) / 18:0] and GlcCer[d18:2(4E,8Z) / 26:0]. 4. 4. An agent for activating the antigen presentation ability of dendritic cells, containing GlcCer[d18:2(4E,8Z) / 18:0] as an active ingredient. 5. An immunostimulant, containing at least one active ingredient selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0]. 6. An immunostimulating food or beverage composition, containing at least one active ingredient selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0]. 7. 7. An immunopotentiating pharmaceutical composition containing as an active ingredient at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0]. A method for stimulating human immunity, comprising: (a) isolating at least one compound selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0] from a rice bran extract; and (b) administering an appropriate amount of the compound isolated in step (a) to a human by oral or parenteral administration, thereby promoting IL-6 production in dendritic cells of the human.

[0008] The present invention relates to a method for enhancing immune responses by using at least one ceramide selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0], which promotes IL-6 production in dendritic cells, thereby enhancing the chemotactic activity of neutrophils toward pathogens and the phagocytic activity of macrophages. IL-6 production is mediated by binding to TLR2 and / or TLR4. TLR2 recognizes lipoproteins, which are components of peptidoglycan contained in the cell walls of Gram-positive bacteria. Thus, GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0] have immunostimulatory effects against infections caused by Gram-positive bacteria. Examples of infections caused by Gram-positive bacteria include, but are not limited to, anthrax, diphtheria, enterococcal infection, erysipelothricosis, listeriosis, pneumococcal infection, Staphylococcus aureus infection, streptococcal infection, and toxic shock syndrome. Furthermore, TLR4 binds to lipopolysaccharide, which constitutes the cell surface of Gram-negative bacteria. This allows TLR4 to have immunostimulatory effects against infections caused by pathogens containing lipopolysaccharide, such as Gram-negative bacteria. Examples of infections caused by Gram-negative bacteria include, but are not limited to, infections caused by Escherichia coli, Salmonella, and Pseudomonas aeruginosa. Furthermore, GlcCer[d18:2(4E,8Z) / 18:0] and GlcCer[d18:2(4E,8Z) / 26:0] have the effect of activating the proliferation of helper T cells by dendritic cells. As a result, GlcCer[d18:2(4E,8Z) / 18:0] and GlcCer[d18:2(4E,8Z) / 26:0] have immunostimulatory effects. Furthermore, GlcCer[d18:2(4E,8Z) / 18:0] has the effect of activating the antigen-presenting ability of dendritic cells.

[0009] 1 is a graph showing the results of an evaluation of the effect on the IL-6 production ability of iMylc cells (immature myeloid cells obtained by inducing differentiation from iPS cells).

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[0010] The immunostimulating agent of the present invention is characterized by comprising, as an active ingredient, at least one ceramide selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0].

[0011] The above ceramides are represented by the following chemical formulas (1) to (4), respectively.

[0012] The method for producing the ceramides is not particularly limited. The ceramides can be obtained by extracting rice bran with a polar solvent to obtain a rice bran extract, and then isolating the ceramides from the extract.

[0013] The "rice bran extract" is not particularly limited as long as it is produced during the process of producing rice bran oil. For example, the rice bran extract can be obtained by extracting rice bran with an organic solvent and then precipitating the extract, but this method is not limited to this. Furthermore, the rice bran extract may be obtained by extracting by-products produced during the production process from rice bran oil.

[0014] The method for isolating the ceramides from the rice bran extract is not particularly limited, and they can be isolated, for example, by chromatography using activated clay, activated carbon, silica gel, alumina, diatomaceous earth, synthetic adsorbents, ion exchange resins, etc., or by removing components other than ceramides by adsorption, decomposition, precipitation, filtration, dissolution, distillation, etc. Note that these compounds may also be obtained by using commercially available rice-derived ceramide extracts, such as Oryza Ceramide (registered trademark) manufactured by Oryza Oil & Fat Chemical Co., Ltd.

[0015] The immunostimulant of the present invention can be used as an ingredient in various foods and beverages. Examples of foods and beverages include general foods such as confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy candy, tablet candy, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, green tea, carbonated drinks, sports drinks, etc.), health foods (tablets, capsules, etc.), and nutritional supplements (nutritional drinks, etc.). The agent of the present invention can be appropriately incorporated into these foods and beverages.

[0016] These foods and beverages can contain various ingredients depending on the type, and the following food ingredients can be used: glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorings, flavorings, and preservatives.

[0017] As a specific manufacturing method, the agent of the present invention is spray-dried or freeze-dried together with powdered cellulose, and then formed into a powder, granules, tablets, or solution, which can be easily incorporated into foods and beverages (instant foods, etc.). The agent of the present invention can also be dissolved in, for example, fats and oils, ethanol, glycerin, or a mixture thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, it can also be mixed with a binder such as gum arabic or dextrin to form a powder or granules, which can then be added to beverages or solid foods.

[0018] When the agent of the present invention is applied to food and drink, the amount of the active ingredient added is preferably 1 to 20 wt % in total relative to the food and drink, since the main purpose is to prevent disease and maintain health.

[0019] The immunopotentiator of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs). Pharmaceutical preparations can be produced by appropriately blending the agent of the present invention with raw materials for pharmaceutical preparations. Examples of pharmaceutical raw materials that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearate monoglyceride, starch, lactose, powdered gum arabic, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), and lubricants (purified talc, stearates, polyethylene glycol, etc.).

[0020] The immunostimulant of the present invention can generally be administered orally in the form of tablets, pills, soft or hard capsules, fine granules, powders, granules, liquids, etc., but may also be administered parenterally. When administered parenterally, it can be administered in the form of a solution or with the addition of a dispersant, suspending agent, stabilizer, etc., by local tissue administration, intradermal, subcutaneous, intramuscular, or intravenous injection. It may also be in the form of a suppository, etc. Furthermore, it can be administered as an eye drop.

[0021] The dosage may vary depending on factors such as the administration method, the condition, and the patient's age. Adults typically receive 0.5 to 5,000 mg of the active ingredient per day, while children typically receive 0.5 to 3,000 mg. The proportion of the immunostimulant can be adjusted depending on the dosage form, but typically, it is approximately 0.3 to 15.0 wt% for oral or mucosal administration and 0.01 to 10 wt% for parenteral administration. The dosage varies depending on various conditions, and therefore, a smaller amount than the above may be sufficient, or it may be necessary to administer a larger amount than the above range.

[0022] Examples of the present invention will be described below. Note that the examples shown below are provided to confirm the various actions and effects of the agent of the present invention obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0023] Example: Preparation of Ceramides (1) Preparation of Glucosylceramides. Oryza ceramide (Oryza Oil & Fat Chemical Co., Ltd.) was crudely fractionated by medium-pressure preparative liquid chromatography. The fractionation conditions were a Yamazen Universal Silica Gel Column Premium, with elution conditions of hexane:ethyl acetate (9:1 → 8:2 → 7:3 → 5:5) → ethyl acetate → chloroform:methanol (9:1 → 8:2 → 7:3 → 5:5) → methanol. The chloroform:methanol (8:2) fraction was then purified by reverse-phase HPLC (COSMOSIL C18 MS-II, methanol) to isolate four glucosylceramides. The purified glucosylceramides were identified as the glucosylceramides represented by the chemical formulas (1), (2), (3), and (5) by comparing their NMR and MS spectra with the literature values ​​listed below. 1. Inagaki. et al., Chem. Pharm. Bull., 52(11), 1307-1311 (2004) 2. Jung JH et al., J. Nat. Prod., 59, 319-322 (1996) 3. Liu H. et al., Phytochemistry., 49(8), 2403-2408 (1998) 4. Ryu J. et al., Arch. Pharm. Res., 26(2), 138-142 (2003) 5. Pittaya T. et al., Chem. Pharm. Bull., 52(1), 27-32 (2004) 6. Kang SS et al., Chem. Pharm. Bull., 49, 321-323 (2001) 7. Luo Y. et al., Lipid, 39(9), 907-914 (2004) 8. Zhang WK ​​et al., Chem. Phys. Lipids., 148, 77-83 (2007) The compound represented by chemical formula (5) is as follows.

[0024] (2) Preparation of the compound (Cer[t18:0 / 22:0]) represented by the above chemical formula (4) Ethanol extract of rice bran was used as the starting material. The extract (10 g) was adsorbed onto silica gel (22 g) to prepare "mabushi." This was fractionated by medium-pressure flash chromatography (Yamazen Corporation). Specifically, "mabushi" was applied to a medium-pressure silica gel flash column (Universal Column Silica Gel, 3 L) and fractionated sequentially with the following solvents (1. hexane:ethyl acetate, 2. chloroform:methanol). An ELSD detector was used for detection.

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[0026] Fractions 17 and 18 were concentrated from the crude fractions eluted with chloroform:methanol (90:10 → 80:20) (5–10 min). These were repeatedly purified by preparative HPLC (C18, Osaka Soda, CAPCELL PAK C18, 20 × 250 mm) using THF:methanol (1:9) as a solvent. The proton and carbon NMR and mass spectra of the resulting compound were compared with those reported in the literature (Gao JM et al., Chem. Phys. Lipids 2004, 131, 205–213). The compound was identified as Cer[t18:0 / 22:0], the compound represented by formula (4).

[0027] Test Example 1: Evaluation of IL-6 production ability in immature dendritic cells (1) Test method In this experiment, iMylc® cells (immature myeloid cells obtained by inducing differentiation from iPS cells) purchased from Mican Technology, Inc. were used. These cells were cultured and evaluated using iMylc® dedicated maintenance medium provided by Mican Technologies. iMylc® cells were seeded into a 96-well plate (1.0 × 104 cells / well / 100 μL), and medium containing the test substance was added (100 μL / well). The cells were then cultured at 37°C in the presence of 5% CO2 for 20 hours. The final DMSO concentration was adjusted to 0.1% (v / v). After the culture, the cells were assayed using a commercially available kit (Human IL-6 ELISA MAX TMThe amount of IL-6 produced in the culture supernatant was measured using a standard kit (Deluxe Set (BioLegend)) according to the standard method. The results are shown in Figure 1.

[0028] (2) Results and Effects of Examples in Test Example 1 As a result of evaluating the immunostimulatory effects of various GlcCer and Cer on dendritic cells, GlcCer[d18:2(4E,8Z) / 18:0] was found to have a significant effect of increasing IL-6 production. Furthermore, compared with the chemical structure of GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], which has a fatty acid carbon number of 26, showed significant activity. Furthermore, significant activity was also observed in GlcCer[t18:1(8Z) / 20:0], which has a different sphingoid base structure, and Cer[t18:0 / 22:0], which has no sugar bound to it. These results demonstrate that GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0] have the ability to promote IL-6 production in dendritic cells, thereby enhancing the chemotactic ability of neutrophils toward pathogens and the phagocytic ability of macrophages, thereby strengthening the immune response. These findings confirm their usefulness as immunostimulants.

[0029] Test Example 2: Evaluation of pattern recognition receptor (PRR) binding activity in dendritic cells (1) Test method The test was performed using iMylc® with various TLR or CLR inhibitors. The cells were cultured and evaluated using iMylc® dedicated maintenance medium provided by Mican Technologies. iMylc® cells were seeded in a 96-well plate (1.0 × 10 4After adding medium containing various inhibitors (see below) (50 μL / well), the cells were cultured at 37°C in the presence of 5% CO2 for 4 hours. Subsequently, medium containing GlcCer[d18:2(4E,8Z) / 18:0] was added (50 μL / well), and the cells were cultured at 37°C in the presence of 5% CO2 for 20 hours. The final DMSO concentration was adjusted to 0.1% (v / v). After the culture, the amount of IL-6 produced in the culture supernatant was measured according to the standard method using a commercially available kit (Human IL-6 ELISA MAX™ Deluxe Set (BioLegend)) (N=4). The results are shown in Figure 3. <List of inhibitors> - TLRs inhibitor: Chloroquine diphosphate (Fujifilm Wako Pure Chemical Industries) - TLR2 inhibitor: MMG-11 (MedChemExpress) - TLR4 inhibitor: LPS-RS (InvivoGen) - TLR3, 7, 9 inhibitor: ODN2088 (InvivoGen) - TLR8 inhibitor: CU-CPT9a (InvivoGen) - Dectin 1 inhibitor: Laminarin (Sigma-Aldrich) - DC-SIGN inhibitor: Fucosyllactose Laminarin (Sigma-Aldrich) - Mincle expression inhibitor: Isoliquiritigenin (Tokyo Chemical Industry Co., Ltd.) The above inhibitors were tested at concentrations that did not cause cytotoxicity in iMylc (registered trademark) cells.

[0030] (2) Results and Effects of Examples in Test Example 2 As a result, IL-6 production by GlcCer[d18:2(4E,8Z) / 18:0] was inhibited by all TLR inhibitors in a concentration-dependent manner, and at high concentrations (20 μM), inhibition of nearly 90% or more was observed. Furthermore, it was significantly inhibited by TLR2 and 4 inhibitors. In summary, these results demonstrate that GlcCer[d18:2(4E,8Z) / 18:0] binds to TLR2 and 4 on dendritic cells, promoting IL-6 production (immune enhancement). (Figure 3) In addition to GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0] are also thought to promote IL-6 production (immune enhancement) through a similar mechanism.

[0031] Test Example 3: Evaluation of Dendritic Cell-Induced Helper T Cell Proliferation (1) Test Method The test was performed using a mixed lymphocyte reaction (MLR). Human peripheral blood-derived αMylc® cells were treated with GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[t18:1(8Z) / 20:0], GlcCer[d18:2(4E,8Z) / 20:0], or GlcCer[d18:2(4E,8Z) / 26:0] (1 or 10 μM) for 24 hours. During this time, peripheral blood-derived mononuclear cells (PBMCs) prepared from human blood were stained with carboxycein diacetate succinimidyl ester (CFDE-SE), a cell membrane-permeable dye, for 7 minutes. After that, the reacted αMylc® cells and PBMCs were mixed at a ratio of 1:2 (2 × 10 4 cells: 4×10 4 The cells were mixed with CD4 (APC staining, helper T cell marker) and co-cultured for 7 days. Afterwards, the mean fluorescence intensity (MFI) of CD4 (APC staining, helper T cell marker) and CFDA (cell proliferation) was quantified using flow cytometry (N=3). The activation index of the MLR in this study was the cell proliferation rate of helper T cells, and the method used is shown in Figure 4.

[0032] (2) Results and Effects of Examples in Test Example 3 The results of Test Example 3 are shown in Figure 5. As shown in Figure 5, 1 µM GlcCer[d18:2(4E,8Z) / 18:0] and 10 µM GlcCer[d18:2(4E,8Z) / 26:0] were found to have a significant effect of increasing the T cell proliferation rate. These results demonstrate that GlcCer enhances the antigen-presenting ability of dendritic cells.

[0033] Test Example 4: Evaluation of activation of antigen-presenting ability of dendritic cells (1) Test method Human peripheral blood-derived αMylc® cells were used in the experiment. αMylc® cells were seeded in a 6-well plate (1.0 × 10 5 After adding medium containing GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[t18:1(8Z) / 20:0], GlcCer[d18:2(4E,8Z) / 20:0], or GlcCer[d18:2(4E,8Z) / 26:0] (1000 μL / well), the cells were cultured for 24 hours at 37°C in the presence of 5% CO2. The final DMSO concentration was adjusted to 0.1% (v / v). After culture, the cells were stained with the following antibodies according to flow cytometry, and the MFI of the cell membrane surface markers was measured using a flow cytometer (N=3). The results are shown in Figure 6, and the results of pseudocolor staining of CD40 and CD86 are shown in Figure 7. <Antibodies> ・PE-CD40 (BioLegend) ・FITC-CD80 (BioLegend) ・FITC-CD86 (BioLegend)

[0034] (2) Results and Effects of the Example in Test Example 4 Figure 6 shows that GlcCer[d18:2(4E,8Z) / 18:0] significantly increased the expression of CD80 and CD40. An increasing trend was also observed for CD86. Thus, Figure 7 shows the pseudocolor results for CD40 and CD86, and it can be seen that the plots for GlcCer[d18:2(4E,8Z) / 18:0] are generally shifted to the upper right compared to the control. This confirms that GlcCer[d18:2(4E,8Z) / 18:0] has the effect of activating the antigen-presenting ability of dendritic cells.

[0035] The following are examples of formulations of the agent (ceramides) according to the present invention. Note that the following formulation examples do not limit the present invention. Formulation Example 1: Chewing gum Sugar 53.45 wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Flavoring 0.5 Ceramides 0.05 100.0 wt%

[0036] Formulation example 2: Gummy Reduced starch syrup 40.9 wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Water 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Coloring 0.02 Ceramides 0.1 100.0 wt%

[0037] Formulation Example 3: Candy Sugar 50.36 wt% Starch syrup 33.0 Water 14.4 Organic acid 2.0 Flavoring 0.2 Ceramides 0.04 100.0 wt%

[0038] Formulation example 4: Yogurt (hard / soft) Milk 41.5 wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 Ceramides 0.04 Flavoring trace Water balance 100.0 wt%

[0039] Formulation example 5: Soft drink Fructose glucose syrup 30.0 wt% Emulsifier 0.5 Ceramides 0.03 Flavoring appropriate amount Purified water Remaining 100.0 wt%

[0040] Formulation Example 6: Tablets Sugar 76.4 wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Ceramides 0.05 Purified water 4.35 100.0 wt%

[0041] Formulation Example 7: Soft capsule Brown rice germ oil 47.9 wt% Yuzu seed oil 40.0 Emulsifier 12.0 Ceramides 0.1 100.0 wt%

[0042] Formulation Example 8: Tablet Lactose 54.9 wt% Microcrystalline cellulose 30.0 Starch hydrolysate 10.0 Glycerin fatty acid ester 5.0 Ceramides 0.1 100.0 wt%

[0043] As described above, the present invention can provide an immunostimulant containing a novel component as an active ingredient.

Claims

1. An agent for promoting IL-6 production in dendritic cells, comprising as an active ingredient at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0].

2. The IL-6 production promoter according to claim 1, characterized in that IL-6 production is achieved by binding to TLR2 and / or TLR4.

3. A dendritic cell-mediated helper T cell proliferation agent comprising at least one active ingredient selected from GlcCer[d18:2(4E,8Z) / 18:0] and GlcCer[d18:2(4E,8Z) / 26:

0.

4. An agent for activating the antigen-presenting ability of dendritic cells, the active ingredient of which is GlcCer[d18:2(4E,8Z) / 18:0].

5. GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0] and Cer[t18:0 / 22: 0]. An immunostimulant comprising as an active ingredient at least one selected from the above.

6. An immunostimulating food or beverage composition containing as an active ingredient at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0], and Cer[t18:0 / 22:0].

7. An immunostimulating pharmaceutical composition comprising as an active ingredient at least one selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0] and Cer[t18:0 / 22:0].

8. A method for stimulating human immunity, comprising the steps of: (a) isolating at least one species selected from GlcCer[d18:2(4E,8Z) / 18:0], GlcCer[d18:2(4E,8Z) / 26:0], GlcCer[t18:1(8Z) / 20:0] and Cer[t18:0 / 22:0] from an extract of rice bran; and (b) orally or parenterally administering to a human an appropriate amount of the compound isolated in the step (a), thereby promoting IL-6 production in dendritic cells of the human.

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