Composition for plasmacytoid dendritic cell activation

Abstract

To provide a composition capable of maintaining, improving, or enhancing immune function by activating plasmacytoid dendritic cells (pDCs).SOLUTION: Lactoferrin and / or a lactoferrin hydrolysate is incorporated as an active ingredient into a composition for pDC activation. Here, the pDC activation can include an increase in one or more selected from the group consisting of the proportion of pDC counts among mononuclear cells, the proportion of CD86+ cell counts among pDCs, the CD86 expression level in pDCs, and the HLA-DR expression level in pDCs. The composition can be consumed in the form of a food / beverage or a pharmaceutical.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for activating plasmacytoid dendritic cells, which contains lactoferrin and / or lactoferrin hydrolysate as an active ingredient. [Background technology]

[0002] Dendritic cells (DCs) are immune cells that highly regulate the innate and adaptive immune systems. Plasmacytoid dendritic cells (pDCs), a type of dendritic cell, express Toll-like receptors (TLR) 7 and TLR 9. pDCs express TLR-7 and TLR-9, recognize viral RNA and DNA, and produce large amounts of type I interferon (IFN) that inhibits viral proliferation. pDCs also act as antigen-presenting cells and activate specific T cells.

[0003] Conventionally, there has been much interest in enhancing immune function and increasing virus defense activity, and components that produce IFN have been studied. For example, Patent Document 1 discloses that lactic acid bacteria can activate pDC and induce IFN production.

[0004] Furthermore, the relationship between lactoferrin (hereinafter also referred to as "LF"), a milk protein, and IFN production has also been reported. Non-Patent Document 1 reports that the ingestion of 1 g of LF per day tends to increase the IFN-α production ability of pDCs. Non-Patent Document 2 reports that the ingestion of 270 mg / day of liposomal LF significantly increases the IFN-α production ability of healthy adult males in a comparison of before and after. In addition, pDCs, which act as antigen-presenting cells, express costimulatory molecules such as CD86 on the cell surface during activation, and Non-Patent Document 3 reports that the ingestion of 300 mg / day of enteric LF significantly increases the number of cells expressing CD86 in healthy elderly people. However, CD86 is generally expressed in antigen-presenting cells such as myeloid dendritic cells, macrophages, and monocytes, and it is not known that lactoferrin increases the number of pDCs or promotes the expression of CD86, a cell surface marker for pDCs, which are a minor population that accounts for only 0.2-0.5% of peripheral blood mononuclear cells (Non-Patent Document 4). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2017-201984 A [Non-patent literature]

[0006] [Non-Patent Document 1] M. v. Splunter et al., Front. Immunol., 2018;9:2677. [Non-Patent Document 2] A. Ishikado et al., Biofactors. 2004;21:69-72. [Non-Patent Document 3] H. Kawakami et al., Int. Dairy J. 2015;47:79-85. [Non-Patent Document 4] Henriquez JE et al., J Acquir Immune Defic Syndr. 2017;75:588-596. Summary of the Invention [Problem to be solved by the invention]

[0007] An objective of the present invention is to provide a composition capable of maintaining, improving, or enhancing immune function by activating pDC. [Means for solving the problem]

[0008] As a result of intensive research aimed at solving the above problems, the present inventors found that when mononuclear cells collected from human peripheral blood are cultured in the presence of LF, pDCs and CD86-positive cells are increased, thereby completing the present invention.

[0009] That is, a first aspect of the present invention is a composition for activating plasmacytoid dendritic cells (pDC), which contains lactoferrin and / or a lactoferrin hydrolysate. The activation may be an increase in one or more selected from the group consisting of the proportion of pDCs among mononuclear cells, the proportion of CD86-positive cells among pDCs, the expression level of CD86 in pDCs, and the expression level of HLA-DR in pDCs. A second aspect of the present invention is a composition for increasing pDCs, which contains lactoferrin and / or a lactoferrin hydrolysate. A third aspect of the present invention is a composition for promoting CD86 and / or HLA-DR expression in pDC, which contains lactoferrin and / or a lactoferrin hydrolysate. The compositions of the present invention are preferably used for maintaining, improving, or enhancing immune function. The compositions of the present invention may be in the form of a food or drink, or a pharmaceutical product. Effect of the Invention

[0010] According to the present invention, immune function can be maintained, improved, or enhanced by activating pDC. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The present invention will now be described in detail. However, the present invention is not limited to the following embodiments and can be freely modified within the scope of the present invention.

[0012] The composition of the present invention contains lactoferrin and / or lactoferrin hydrolysate as an active ingredient.

[0013] Lactoferrin is an iron-binding glycoprotein found in the milk, tears, saliva, blood, etc. of mammals, such as sheep, goats, pigs, mice, buffalo, camels, yaks, horses, donkeys, llamas, cows, and humans. The lactoferrin in the present invention may be derived from any mammal and is not particularly limited, but in terms of content and availability, lactoferrin derived from milk of, for example, cows, humans, etc. The milk may be any of colostrum, transitional milk, normal milk, and terminal milk. The lactoferrin in the present invention may be lactoferrin separated by a standard method (e.g., ion chromatography) from the processed milk such as skim milk or whey, recombinant lactoferrin produced by genetic engineering from microorganisms, animal cells, transgenic animals, or synthetic lactoferrin, or a mixture thereof. Lactoferrin may be non-glycosylated or glycosylated. As such lactoferrin, commercially available lactoferrin produced on an industrial scale (e.g., manufactured by Morinaga Milk Industry Co., Ltd.) may be used.

[0014] The metal content in lactoferrin in the present invention is not particularly limited, and any one or a mixture of two or more selected from the group consisting of apo-lactoferrin obtained by de-ironizing lactoferrin with hydrochloric acid, citric acid, etc.; metal-saturated lactoferrin with a degree of saturation of 100% or more obtained by chelating the apo-lactoferrin with metals such as iron, copper, zinc, manganese, etc.; and metal-partially saturated lactoferrin in which metals are bound to each other at various degrees of saturation less than 100% can be used.

[0015] Here, an example of a method for preparing lactoferrin (separating and purifying lactoferrin from raw materials such as milk) used in the present technology is shown below, but the present invention is not limited to this. A raw milk material derived from cows is passed through a cation exchange column, the passing liquid is collected, and the passing liquid is passed through the column repeatedly as appropriate. Deionized water and saline are passed through the column to obtain an eluate of basic proteins adsorbed on the cation exchange column. Proteins are collected from the eluate, washed as appropriate, and dissolved in deionized water, and the solution is filtered through an ultrafiltration membrane. Powdered lactoferrin is obtained by further desalting and freeze-drying.

[0016] More specifically, first, the ion exchanger is packed in a column, hydrochloric acid is passed through, and the ion exchanger is equilibrated by washing with water. Then, skim milk with a pH of 6.9 cooled to 4°C is passed through the column, the permeate is collected, and the column is passed through again in the same manner. Next, deionized water is passed through the column, and saline is passed through the column to obtain an eluate of basic proteins adsorbed on the ion exchanger. The proteins recovered from the eluate are washed, dissolved by adding deionized water, and the resulting solution is desalted using an ultrafiltration membrane module and freeze-dried to obtain powdered bovine lactoferrin. In this way, bovine lactoferrin with a purity of 95% by mass or more is obtained.

[0017] In lactoferrin, depending on the difference of species, genus, individual, etc., there are naturally genetic mutations such as substitution, deletion, insertion, addition, or inversion of one or more bases at one or more positions, and the amino acid of the protein encoded by the gene having such mutation may also be mutated. The lactoferrin that can be used in the present technology of the present invention also includes those containing such mutations, as long as the effect of the present invention is not impaired. Furthermore, lactoferrin in the present invention may also include lactoferrin that has been treated with heat, acid, or alkali, so long as the effects of the present invention are not impaired.

[0018] The lactoferrin hydrolysate of the present invention refers to a product obtained by hydrolyzing the above-mentioned lactoferrin. An example of the hydrolysis treatment is the method described in JP-A-2012-235768. Specifically, the pH of the lactoferrin solution is adjusted to 2 to 4, preferably 2.5 to 3.5, and particularly preferably pH 3, with an acid such as hydrochloric acid, citric acid, or acetic acid before the enzyme reaction treatment. A desired amount of protease is added to the pH-adjusted lactoferrin solution, and the temperature of the enzyme reaction is maintained at 35 to 55°C, preferably 40 to 50°C, and more preferably 42 to 48°C, and the lactoferrin is hydrolyzed with stirring for 6 to 24 hours, preferably 12 to 18 hours. Next, for example, the reaction solution is heated to 80° C. and maintained for 10 minutes to inactivate the enzyme, and then the pH is adjusted to 5 to 7, for example 6, preferably by adding an alkaline solution such as a sodium hydroxide solution. The reaction solution (lactoferrin hydrolysate) after pH adjustment may be left as a solution, but is preferably pulverized by freeze-drying, etc. The lactoferrin hydrolysate may also be fractionated by chromatography, ultrafiltration, or the like.

[0019] In the composition of this embodiment, the content of lactoferrin and / or lactoferrin hydrolysate in the total composition is preferably 0.001% by mass or more and less than 100% by mass, more preferably 0.005 to 95% by mass, and even more preferably 0.01 to 85% by mass.

[0020] The compositions of the present invention are capable of activating pDCs. Here, the activation of pDCs includes an increase in the proportion of pDCs in mononuclear cells, which means that the proportion of pDCs in the total mononuclear cell population per unit of time increases after application of the composition of the present invention compared to before application, or increases when the composition of the present invention is applied compared to when it is not applied. This means that the increase in the ratio may be due to an absolute increase in the number of pDCs. In other words, the composition of the present invention can increase pDCs. It is presumed that the increase in the ratio or absolute number of pDCs by the composition of the present invention is due to the composition of the present invention acting on pDC precursor cells to promote their differentiation into pDCs. The number of pDC cells after application of the composition of the present invention is at least 1.1 times, preferably 1.5 times or more, preferably 2 times or more, preferably 2.5 times or more, preferably 3 times or more, preferably 3.5 times or more, and more preferably 4 times or more, compared to before application. The upper limit is not particularly limited, but may be 8 times or less, 7 times or less, 6 times or less, or 5 times or less. In addition, it may be within a range of a combination of the above numerical values ​​that are not contradictory, specifically, it may be 1.1 to 8 times, 1.5 to 8 times, 2 to 8 times, 2.5 to 8 times, 3 to 8 times, 3.5 to 8 times, or 4 to 8 times.

[0021] Activation of pDC also includes high expression of pDC activation markers. As mentioned above, pDCs also function as antigen-presenting cells, and human pDCs are major histocompatibility complex class II (MHC-II) leukocyte antigen complex (HLA)-DR positive cells. pDCs can activate CD4+ T cells and CD8+ T cells by presenting foreign antigens via HLA-DR. At that time, costimulatory signals from costimulatory molecules such as CD86, CD40, CD80, and CD83 are required for T cell activation. Therefore, high expression of MHC-II HLA-DR and costimulatory molecule CD86 on pDCs is an indicator of high activity of pDCs as antigen-presenting cells (T. S. Mathan et al., Front Immunol., 2013;4:372.; Swiecki M. et al., Nat Rev Immunol., 2015;15:471-485.) 0, CD83, and the like can be used for evaluation as pDC activation markers.

[0022] High expression of pDC activation markers includes an increase in the proportion of CD86-positive cells in pDCs. This means that the proportion of CD86-positive cells in the entire pDC group in any unit increases after application of the composition of the present invention compared to before application, or when the composition of the present invention is applied compared to when it is not applied. The increase in the proportion may be due to an absolute increase in the number of CD86-positive cells. CD86-positive cells refer to cells expressing CD86, and the expression of CD86 in pDCs indicates the activation of pDCs. Note that pDCs are originally HLA-DR-positive cells, and unlike CD86, which is expressed only by a portion of pDCs, the proportion of HLA-DR-positive cells is not an indicator of pDC activation (Dopheide JF et al., Thromb Haemost. 2012;108:1198-207.). Furthermore, high expression of pDC activation markers includes an increase in the amount of CD86 and / or HLA-DR expressed in pDC. This means that the amount of CD86 and / or HLA-DR expressed per unit pDC (in other words, per pDC cell) increases after application of the composition of the present invention compared to before application, or when the composition of the present invention is applied compared to when it is not applied. In other words, the composition of the present invention can promote the expression of CD86 and / or HLA-DR in pDC. Here, the expression of CD86 and / or HLA-DR can be confirmed qualitatively or quantitatively by known methods, for example, immunological techniques using fluorescently labeled antibodies.

[0023] Since the composition of the present invention activates pDCs, it is expected to activate the functions of various immune cells and promote IFN production, and therefore it is expected to maintain, improve, or strengthen immune function. "Maintaining immune function" includes maintaining a state in which normal or good immune function is being exerted and preventing the function from declining, and "improving immune function" includes restoring a declined immune function to a normal state.

[0024] The subject to which the composition of the present invention is administered (ingested) is not particularly limited as long as it is an animal, The subject is usually a mammal, preferably a human, and is not particularly limited, but is preferably a healthy individual, where a healthy individual refers to an individual not suffering from any disease or illness.

[0025] Another aspect of the invention is the use of lactoferrin and / or lactoferrin hydrolysate in the manufacture of a composition for activating pDCs. Another aspect of the invention is the use of lactoferrin and / or lactoferrin hydrolysates in pDC activation. Another aspect of the invention is lactoferrin and / or lactoferrin hydrolysates for use in pDC activation. Another aspect of the invention is a method of activating pDCs comprising administering lactoferrin and / or a lactoferrin hydrolysate to a subject.

[0026] Another aspect of the present invention is the use of lactoferrin and / or lactoferrin hydrolysate in the manufacture of a composition for expanding pDCs. Another aspect of the invention is the use of lactoferrin and / or lactoferrin hydrolysates in expanding pDCs. Another aspect of the invention is lactoferrin and / or lactoferrin hydrolysate for use in expanding pDCs. Another aspect of the invention is a method of increasing pDCs comprising administering lactoferrin and / or lactoferrin hydrolysate to a subject.

[0027] Another aspect of the present invention is the use of lactoferrin and / or a lactoferrin hydrolysate in the manufacture of a composition for promoting CD86 and / or HLA-DR expression in pDC. Another aspect of the present invention is the use of lactoferrin and / or lactoferrin hydrolysate in promoting CD86 and / or HLA-DR expression in pDC. Another aspect of the present invention is lactoferrin and / or a lactoferrin hydrolysate used to promote CD86 and / or HLA-DR expression in pDCs. Another aspect of the present invention is a method for promoting expression of CD86 and / or HLA-DR in pDCs, comprising administering lactoferrin and / or a lactoferrin hydrolysate to a subject.

[0028] In addition, "administering lactoferrin and / or lactoferrin hydrolysate to a subject" may be synonymous with "allowing a subject to ingest lactoferrin and / or lactoferrin hydrolysate." Ingestion may be voluntary (free intake) or forced (forced intake). That is, the administration step may specifically be, for example, a step of blending lactoferrin and / or lactoferrin hydrolysate with food, drink, or feed and supplying it to the subject, thereby allowing the subject to freely ingest lactoferrin and / or lactoferrin hydrolysate.

[0029] The timing and duration of ingestion (administration) of the composition of the present invention are not particularly limited and can be appropriately selected depending on the condition of the subject to be administered.

[0030] The composition of the present invention may be in the form of a food, drink, medicine, etc. by itself, or may be in the form of being contained in a food, drink, medicine, etc. as an additive. The composition of the present invention may be administered orally or parenterally, but is usually administered orally. Parenteral administration includes transdermal, intravenous, rectal, and inhalation.

[0031] The content of the composition of the present invention when it is orally taken (administered) may be the content in the entire composition described above, or it may be appropriately diluted. For example, The content of lactoferrin and / or lactoferrin hydrolysate in the total composition is preferably 0.001 to 100% by mass, more preferably 0.005 to 95% by mass, and even more preferably 0.01 to 85% by mass. These contents may be within the range generally used when distributed as oral compositions.

[0032] The intake (administration) amount of the composition of the present invention is appropriately selected depending on the age (months), sex, condition, other conditions, etc. of the subject to be ingested (administered). The intake (administration) amount of the composition of the present invention is, for example, preferably 0.01 to 1000 mg / kg / day, more preferably 0.1 to 500 mg / kg / day, and even more preferably 1 to 300 mg / kg / day, in terms of the intake amount of lactoferrin and / or lactoferrin hydrolysate. It is advisable to use the amount within the range as a guide. Regardless of the amount or period of intake (administration), the composition of the present invention can be administered once a day or in divided doses.

[0033] When the composition of the present invention is intended to be orally ingested, it is preferably in the form of a food or drink. There are no particular limitations on the form or properties of the food and beverage products as long as they do not impair the effects of the present invention and can be taken orally. Except for containing lactoferrin and / or lactoferrin hydrolysate, they can be produced by conventional methods using raw materials normally used for food and beverage products.

[0034] Food and drink products include, regardless of their form, liquid, paste, gel, solid, powder, etc., such as tablet confectionery; liquid food (nutritional food for tube feeding); wheat flour products such as bread, macaroni, spaghetti, noodles, cake mix, fried chicken powder, breadcrumbs, etc.; instant noodles, cup noodles, retort / prepared foods, pre-cooked canned foods, microwave foods, instant soups / stews, instant miso soup / cleansing soups, canned soups, freeze-dried foods, other instant foods, etc.; canned agricultural products, canned fruit, jams, etc. Agricultural processed products such as marmalades, pickles, boiled beans, dried agricultural goods, cereals (processed grain products); marine processed products such as canned seafood, fish ham and sausages, seafood paste products, seafood delicacies, tsukudani (fried fish dishes); livestock processed products such as canned livestock pastes, livestock ham and sausages; dairy products such as processed milk, milk drinks, yogurt (fermented milk), lactic acid bacteria drinks, cheese, ice cream, cream, and other dairy products; fats and oils such as butter, margarines, vegetable oils, soy sauce, miso, sauces, Basic seasonings such as processed tomato seasonings, mirin, vinegar, etc.; complex seasonings and foods such as cooking mixes, curry bases, sauces, dressings, noodle soups, spices, and other complex seasonings; frozen foods such as ingredient frozen foods, semi-cooked frozen foods, and cooked frozen foods; confectioneries such as caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pies, snacks, crackers, Japanese sweets, rice sweets, bean sweets, dessert sweets, jellies, and other sweets. ; carbonated drinks, natural fruit juice, fruit juice drinks, soft drinks with fruit juice, fruit pulp drinks, fruit drinks with fruit particles, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic drinks, other beverages, and other commercially available foods such as baby food, furikake, and ochazuke nori; nutritional compositions such as supplements and prepared milk (including powdered milk, liquid milk, etc.); enteral nutritional foods; functional foods (foods for specified health uses, foods with nutritional functions), etc.

[0035] When the food or drink is in the form of a supplement, it can be formulated into solid preparations such as powders, granules, tablets, capsules, etc., which may be enteric-coated, etc.; liquid preparations such as solutions, syrups, suspensions, emulsions, etc. When formulating such preparations, the ingredients, carriers, and methods for formulating pharmaceuticals described below can be followed.

[0036] In addition, one aspect of the food and drink may be feed, such as pet food, livestock feed, and fish feed. The form of the feed is not particularly limited, and may contain, in addition to lactoferrin and / or lactoferrin hydrolysate, for example, grains such as corn, wheat, barley, rye, milo, etc.; vegetable oil cakes such as soybean oil cake, rapeseed oil cake, palm oil cake, linseed oil cake, etc.; bran such as wheat bran, rice bran, defatted rice bran, etc.; manufacturing residues such as corn gluten meal and corn jam meal; animal feeds such as fish meal, skim milk powder, whey, yellow grease, tallow, etc.; yeasts such as torula yeast and brewer's yeast; mineral feeds such as calcium phosphate, calcium carbonate, etc.; oils and fats; simple amino acids; sugars, etc.

[0037] When the composition of the present invention is in the form of a food or drink (including feed), it can be provided and sold as a food or drink labeled with the purpose of maintaining immune function, etc. Furthermore, lactoferrin and / or lactoferrin hydrolysate according to the present specification can be used for the production of such food or drink.

[0038] Such "indication" acts include all acts for informing consumers of the above-mentioned uses, and any expression that can recall or infer the above-mentioned uses falls under the category of "indication" acts in this invention, regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. Furthermore, it is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the above-mentioned uses.Specific examples include the act of transferring, delivering, displaying for the purpose of transferring or delivering, or importing food and beverage products or product packaging on which the above-mentioned uses are written, displaying or distributing advertisements, price lists, or transaction documents related to the products and writing the above-mentioned uses in information containing the above-mentioned uses and providing them by electromagnetic means (such as the Internet), etc.

[0039] On the other hand, the content of the labeling is preferably a labeling approved by the government etc. (for example, a labeling approved based on various systems established by the government and made in a manner based on such approval, etc.) In addition, it is preferable that such a labeling content is affixed to the packaging, containers, catalogs, pamphlets, POP and other sales site promotional materials, other documents, etc.

[0040] In addition, "labeling" also includes labeling as health food, functional food, enteral nutrition food, special purpose food, health functional food, food for specified health uses, nutrient functional food, functional food, functional labeling food, medical drug, etc. Among these, in particular, labeling approved by the Consumer Affairs Agency, for example, labeling approved under the system related to food for specified health uses, nutrient functional food, or functional labeling food, or a similar system, etc., can be mentioned. Specifically, labeling as food for specified health uses, labeling as food for conditional specified health uses, labeling that affects the structure or function of the body, labeling for reducing disease risk, labeling of functionality based on scientific evidence, etc., can be mentioned, and more specifically, labeling as food for specified health uses (especially labeling of health uses) and similar labeling as specified in the Cabinet Office Ordinance on the Permission of Labeling for Special Uses Prescribed in the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009) are typical examples. Examples of such claims include "acts on pDC (plasmacytoid dendritic cells)", "acts on pDC immune cells", "activates pDC immune cells", "increases pDC immune cells", "helps maintain immune function in healthy people", "boosts immune function", "supports immune function", "supports maintaining immune function in healthy people", "acts on pDC (plasmacytoid dendritic cells) and helps maintain immune function in healthy people", etc. can be done.

[0041] The composition of the present invention may also be in the form of a pharmaceutical product. The route of administration of pharmaceuticals may be either oral or parenteral, with oral being preferred. Parenteral intake (administration) includes transdermal, intravenous, rectal, inhalation, etc. The pharmaceutical form can be formulated into a desired dosage form depending on the method of administration. For example, in the case of oral administration, solid preparations such as powders, granules, tablets, and capsules; solutions, capsules, and capsules; The compound can be formulated into liquids such as lops, suspensions, and emulsions. It can also be made into an enteric-coated formulation by enteric coating. For parenteral administration, the compound can be formulated into suppositories, ointments, injections, and the like. In the formulation, in addition to lactoferrin and / or lactoferrin hydrolysate, ingredients that are usually used in formulations, such as excipients, pH adjusters, colorants, flavorings, etc., can be used. It is also possible to use other medicinal ingredients, or ingredients that have a known or future pDC activating effect, in combination. In addition, formulation can be carried out by a known method according to the dosage form. When formulating, a carrier usually used in formulation may be appropriately mixed. Such carriers include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, etc.

[0042] Examples of excipients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose, and calcium carboxymethyl cellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

[0043] Examples of binders include gelatin, polyvinylpyrrolidone, macrogol, and the like, in addition to the above-mentioned excipients.

[0044] Examples of disintegrants include the above-mentioned excipients, as well as chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone.

[0045] Examples of lubricants include talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as pea gum and geranium stearate; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; sodium carboxylates such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; starch derivatives, and the like.

[0046] Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.

[0047] Examples of flavoring agents include sweeteners, acidulants, and fragrances. In the case of a liquid preparation for oral administration, examples of the carrier used include solvents such as water.

[0048] The timing of taking the pharmaceutical agent of the present invention is not particularly limited, and may be before or after a meal, between meals, or before going to bed. EXAMPLES

[0049] The present invention will be described in more detail below using examples, but the present invention is not limited to these examples.

[0050] [Example 1] Mononuclear cells (PBMCs) were collected from human peripheral blood collected from volunteers (n=3). The mice were treated with or without toferin (MLF-2 manufactured by Morinaga Milk Industry Co., Ltd.) (100-200 μg / mL). Control group: RPMI1640 medium supplemented with 5% human serum, Glutamax, and 1% Penicillin Streptomycin. 6 cells / mL, 37°C, 5% CO The cells were cultured under two conditions. 24 hours after the start of culture, the cells were harvested, and the expression of the cell surface marker CD86 of each immune cell in the PBMC was evaluated using a flow cytometer. The expression of CD86 (positive) indicates the activation of pDC. The results are shown in Table 1. In all samples, when cultured in the presence of lactoferrin, the ratio of pDC count to the total number of cells counted and the ratio of CD86-positive cells in pDC were found to increase. In addition, the amount of CD86 expression per pDC cell was also found to increase.

[0051] [Table 1]

[0052] [Example 2] Mononuclear cells (PBMCs) were collected from human peripheral blood collected from volunteers (n=2). Lactoferrin (MLF-2, manufactured by Morinaga Milk Industry Co., Ltd.) (100 μg / mL) was added or not added (control). (control group), 5.0 × 10 cells were cultured in RPMI1640 medium supplemented with 5% human serum, ssRNA (5 μg / mL), Glutamax, and 1% Penicillin Streptomycin. 5 cells / mL The cells were cultured at 37°C and 5% CO2. 24 hours after the start of culture, the cells were harvested and the expression of cell surface markers CD86 and HLA-DR of each immune cell in the PBMCs was evaluated using a flow cytometer. The expression of CD86 and HLA-DR, respectively, indicates the activation of pDCs. The results are shown in Table 2. It was found that when cultured in the presence of lactoferrin, the expression levels of CD86 and HLA-DR per pDC1 cell also increased.

[0053] [Table 2]

[0054] [Production Example 1] As one embodiment of the composition of the present invention, a method for producing fermented milk containing lactoferrin is described below. First, milk ingredients, lactoferrin (Mirai Co., Ltd.), and other ingredients are mixed and dissolved, with water added as necessary, to prepare a milk preparation. Lactoferrin is added so that the content is 100 mg per 100 g of milk preparation. Next, the milk preparation is homogenized and heat sterilized by a conventional method. A lactic acid bacteria starter is added (inoculated) to the heat-sterilized sterilized milk preparation, and the mixture is fermented at a predetermined fermentation temperature to obtain a fermented product. Curd is formed by fermentation. As the lactic acid bacteria starter, for example, lactic acid bacteria commonly used in yogurt production, such as Lactobacillus bulgaricus, Lactococcus lactis, and Streptococcus thermophilus, can be used. When the pH reaches the target value, the formed curd is broken down by stirring and cooled to below 10°C to obtain lactoferrin-containing fermented milk. Since pDCs are also present in the human intestinal tract, ingesting the above-mentioned lactoferrin-containing fermented milk is expected to have the effect of activating plasmacytoid dendritic cells (pDCs), increasing the number of pDCs, and / or promoting the expression of CD86 and HLA-DR in pDCs.

[0055] [Production Example 2] As one embodiment of the composition of the present invention, a method for producing a tablet confectionery containing lactoferrin is described below. The raw materials having the following composition are mixed and compressed into tablets by a conventional method to obtain tablets containing 300 mg of lactoferrin per gram. Bovine lactoferrin (Mirai Co., Ltd.) 30.0 (mass%) Sorbitol (Towa Chemical Industry Co., Ltd.) 18.0 Cornstarch (Kato Chemicals) 13.0 Reduced maltose (Towa Chemical Industry Co., Ltd.) 36.8 Glycerol fatty acid ester (Riken Vitamin Co., Ltd.) 1.8 Flavor (Hasegawa Fragrance Co., Ltd.) 0.4 Ingestion of the lactoferrin-containing tablets has the effect of activating plasmacytoid dendritic cells (pDCs), increasing pDCs, and / or promoting the expression of CD86 and HLA-DR in pDCs. is expected.

Claims

[Claim 1] A composition for activating plasmacytoid dendritic cells (pDC), comprising lactoferrin and / or lactoferrin hydrolysate.

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