WO2017022560A1 - Promoteur de prolifération de cellules dendritiques plasmacytoïdes - Google Patents

Promoteur de prolifération de cellules dendritiques plasmacytoïdes Download PDF

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Publication number
WO2017022560A1
WO2017022560A1 PCT/JP2016/071814 JP2016071814W WO2017022560A1 WO 2017022560 A1 WO2017022560 A1 WO 2017022560A1 JP 2016071814 W JP2016071814 W JP 2016071814W WO 2017022560 A1 WO2017022560 A1 WO 2017022560A1
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WIPO (PCT)
Prior art keywords
agent
lactic acid
culture
lactobacillus helveticus
plasmacytoid dendritic
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PCT/JP2016/071814
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English (en)
Japanese (ja)
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哲弘 小川
知広 細谷
史彦 酒井
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雪印メグミルク株式会社
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Publication of WO2017022560A1 publication Critical patent/WO2017022560A1/fr

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis

Definitions

  • the present invention relates to a plasmacytoid dendritic cell increasing agent, a food and beverage for increasing the number of plasmacytoid dendritic cells, an increase in the number of plasmacytoid dendritic cells, comprising lactic acid bacteria belonging to Lactobacillus helveticus as an active ingredient
  • the present invention relates to a nutritional composition and a feed for increasing the number of plasmacytoid dendritic cells.
  • Immunity is a biological defense mechanism possessed by many animals, and plays a role in eliminating pathogens such as bacteria and viruses that invade from the outside and tumor cells generated in vivo.
  • the innate immune system is an innate immune system
  • the acquired immune system is an immune system that is acquired in response to the stimulation of a foreign substance.
  • foreign substances such as bacteria and viruses enter the body
  • macrophages, NK cells, and dendritic cells which are responsible for the innate immune system, work to remove foreign substances.
  • dendritic cells transmit information on foreign substances to T cells and B cells, thereby inducing CD8 + T cells that specifically attack the foreign substances and antibody production to activate the acquired immune system.
  • the dendritic cell is a cell that is responsible for the innate immune system and that also participates in the acquired immune system, and thus functions as a control tower for the entire immunity.
  • dendritic cells There are various types of dendritic cells, which are roughly classified into classical dendritic cells (conventional dendritic cells) and plasmacytoid dendritic cells (pDC).
  • Classical dendritic cells play an important role in the induction of the acquired immune system because of their strong antigen-presenting ability.
  • Plasmacytoid dendritic cells have a weak antigen-presenting ability, but they regulate not only the immunostimulatory system toward the elimination of foreign substances but also the immune control system that suppresses excessive immune responses, so that the homeostasis of the immune system It is the cell that is the key to keeping it safe.
  • Type I and type III interferon are cytokines that induce the production of antiviral factors that are inherently possessed by living bodies in order to prevent the growth of viruses, and have a role in preventing viral infection from spreading to the entire tissue.
  • production of type I IFN is strongly stimulated by binding of RNA or DNA derived from virus to TLR7 or TLR9 of cells. Therefore, type I IFN inducers that stimulate TLR7 or TLR9 as antiviral agents have been extensively studied so far.
  • the interferon production inducer described in Patent Document 1 contains lactic acid bacteria as an active ingredient.
  • plasmacytoid dendritic cells are activated to induce IFN production, and the present invention shows It is essentially different from those that increase the number of plasmacytoid dendritic cells.
  • Lactococcus lactis is the only lactic acid bacterium that activates plasmacytoid dendritic cells in the patent document, and its activating effect on lactic acid bacteria belonging to the genus Lactobacillus. It is shown that there is no.
  • Lactobacillus helveticus has not been studied at all.
  • a characteristic of plasmacytoid dendritic cells in the immune control system is to induce IL-10-producing T cells and regulatory T cells, which greatly contributes to suppression of inflammatory diseases.
  • the biological defense function is enhanced with an immunostimulant, there is a concern that inflammation may be caused by the excessive ecological defense function.
  • anti-inflammatory agents are used, there has been a concern that the risk of infection will increase due to excessively low biological defense functions.
  • plasmacytoid dendritic cells have functions of both immunostimulants and anti-inflammatory agents and are important defense functions against viral infections, but at the same time have a function to act on the immune control system, which is just an immune function. It is a key cell involved in maintaining homeostasis.
  • the interferon production inducer described in Patent Document 1 has a certain control effect against virus infection.
  • high type I IFN in serum is an exacerbating factor, such as systemic lupus erythematosus. Therefore, when an interferon production inducer such as Patent Document 1 is used, a plasmacytoid tree is more than necessary. It is not preferable because it stimulates dendritic cells to promote production of excess type I IFN. Since a considerable amount of type I IFN is produced from plasmacytoid dendritic cells at the time of virus infection, it is possible to maintain a sufficient number of cells at the time of viral infection rather than to activate plasmacytoid dendritic cells at normal times. Considered important.
  • the present invention provides a drug or nutrition having an action of increasing plasmacytoid dendritic cells, which can bring about effects of both prevention of infectious diseases and prevention of inflammatory diseases by increasing plasmacytoid dendritic cells. It is an object to provide a composition, food and drink, and feed.
  • the present invention has the following constitutions: (1) A plasmacytoid dendritic cell increasing agent comprising a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof as an active ingredient.
  • Plasmacytoid administration of a plasmacytoid dendritic cell increasing agent comprising a lactic acid bacterium belonging to Lactobacillus helveticus or its culture as an active ingredient to a subject having a reduced number of plasmacytoid dendritic cells Use method of dendritic cell increasing agent.
  • a method of using an immunostimulant comprising administering an immunostimulant comprising a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof as an active ingredient to a subject with reduced immunity.
  • (11) Use of a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof in the production of an immunostimulator.
  • (13) A virus infection preventive agent or virus comprising administering a virus infection preventive agent or therapeutic agent comprising a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof as an active ingredient to a virus infection preventive subject or a virus infected subject How to use infection treatment.
  • a method of using an anti-inflammatory agent comprising administering an anti-inflammatory agent comprising a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof as an active ingredient to a subject having inflammation.
  • an anti-inflammatory agent comprising a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof as an active ingredient to a subject having inflammation.
  • Ingestion of Lactobacillus helvetius induces an increase in plasmacytoid dendritic cells in vivo, preventing and improving excessive immune responses in addition to infection prevention and treatment effects against viruses, etc. Can reduce inflammatory effects.
  • plasmacytoid dendritic cells can be increased and the above-mentioned action can be achieved.
  • FIG. 4 is a graph comparing the number of plasmacytoid dendritic cells in mesenteric lymph nodes after oral administration of Lactobacillus helveticus strain SBT2171 cells to mice.
  • FIG. 4 is a graph comparing the numbers of plasmacytoid dendritic cells in mesenteric lymph nodes after oral administration of a bacterial cell culture of Lactobacillus helveticus strain SBT2171 to mice.
  • a lactic acid bacterium belonging to Lactobacillus helveticus can be used as a lactic acid bacterium in the present invention.
  • Lactobacillus helveticus SBT2171 strain (deposited as FERM BP-5445), SBT-2161 strain (deposited as NITE ABP-01707), SBT2195 strain (deposited as FERM P-11538), SBT2196 strain (FERM P -11676), SBT0064 strain (deposited as FERM P-21079), SBT0402 strain (deposited as FERM P-21559) and the like are preferred, but not limited thereto.
  • Lactobacillus helveticus can be cultured according to a conventional method for lactic acid bacteria culture.
  • various media such as a milk medium or a medium containing milk components and a semi-synthetic medium not containing the milk medium can be used.
  • examples of such a medium include reduced skim milk medium.
  • the cells isolated from the obtained culture by means of collection such as centrifugation can be used as they are as the active ingredient of the present invention. Concentrated, dried, freeze-dried cells, etc. can be used, or dead cells can be formed by heat drying. Not only those that are purely isolated as cells, but also cultures, suspensions, other cell-containing materials, and cytoplasm and cell wall fractions obtained by treating cells with enzymes or physical means should be used. it can.
  • Examples of the culture form include not only a culture medium using a medium generally used for cultivation of lactic acid bacteria, such as MRS medium (manufactured by DIFCO), which is a synthetic medium, and reduced skim milk medium, but also cheese, fermented milk, milk.
  • MRS medium manufactured by DIFCO
  • dairy products such as product lactic acid bacteria drink
  • a culture supernatant prepared by removing milk protein precipitates and bacterial cell components from the obtained culture using methods such as centrifugation and filtration can also be used. Since it is a supernatant with a low solid content, the range of application to foods and drinks is widened.
  • the culture supernatant can be prepared by centrifuging the reduced skim milk culture at 5,000 rpm for 10 minutes.
  • the action of increasing plasmacytoid dendritic cells in the present invention refers to an action of increasing the number of plasmacytoid dendritic cells. By increasing the number of plasmacytoid dendritic cells, the functions of the immunostimulant and anti-inflammatory agent possessed by the cells can be exhibited.
  • excipients, stabilizers, flavoring agents and the like that are permitted in the formulation are mixed as appropriate and concentrated, freeze-dried, or heat dried to form dead cells. These dried products, concentrates, and pastes are also included.
  • excipients, binder, disintegrant, lubricant, flavoring agent, suspending agent, coating agent, and any other optional agent it can also be formulated by mixing drugs.
  • the dosage form can be tablets, capsules, granules, powders, powders, syrups, etc., and these are preferably administered orally.
  • the plasma cell-like dendritic cell increasing agent of the present invention may be added to any food or drink, and may be added to the raw material during the production process of the food or drink.
  • food and drink include dairy products such as cheese, fermented milk, dairy lactic acid bacteria beverages, lactic acid bacteria beverages, butter and margarine, beverages such as dairy beverages, fruit juice beverages and soft drinks, eggs such as jelly, candy, pudding and mayonnaise Processed products, confectionery and breads such as butter cake, and various types of powdered milk, infant foods, nutritional compositions and the like can be mentioned, but are not particularly limited.
  • the plasma cell-like dendritic cell-increasing agent of the present invention can be used as an immunostimulant, a viral infection preventive or therapeutic agent, an immune tolerance inducer, or an anti-inflammatory agent containing this as an active ingredient.
  • the plasmacytoid dendritic cell increasing agent of this invention can be mix
  • the blending ratio of Lactobacillus helveticus to each material is not particularly limited, and is easy to manufacture and preferable What is necessary is just to adjust suitably according to a daily dose.
  • the dose is individually determined in consideration of the symptom, age, etc. of the subject of administration, but in general, in the case of an adult, 10 to 200 g / day of the Lactobacillus helveticus culture, or the fungus body itself, is used. What is necessary is just to adjust a compounding quantity so that 0.1-5,000 mg / day can be ingested. A desired effect can be exhibited by ingesting in this way.
  • FACS fluorescence activated cell sorting
  • Example 1 Production of Lactobacillus helveticus culture
  • Raw milk was sterilized by heating (75 ° C, 15 seconds), cooled to 30 ° C, and 0.01% calcium chloride was added.
  • 0.7% of commercially available lactic acid bacteria starter (LD starter, Christian Hansen) and 1% of Lactobacillus helveticus strain SBT2171 (FERM BP-5445) are added, and 0.003% of rennet is further added. Solidified.
  • the curd thus obtained was cut and stirred until the pH was 6.2 to 6.1, and the whey was discharged to obtain curd grains. Then, the curd grains were packed and pressed, further salted, and aged at 10 ° C.
  • Test Example 2 Oral administration test of Lactobacillus helveticus culture to mice Test Method An oral administration test was performed using the culture of Lactobacillus helveticus SBT2171 obtained in Example 1. The test method was performed according to Test Example 1. The ratio of plasmacytoid dendritic cells to the total number of cells in the mesenteric lymph nodes after dissection was measured, and the results are shown in FIG. 2. 2. Test results From FIG. 2, the trait in the mesenteric lymph node in the group (LH2171 group) ingesting the diet to which the culture of Lactobacillus helveticus SBT2171 was added was greater than in the group ingesting the diet not added (control group). It was found that the proportion of cell-like dendritic cells was significantly higher. That is, it was shown that the number of plasmacytoid dendritic cells increased by ingesting the Lactobacillus helveticus culture.
  • Example 2 Manufacture of a plasmacytoid dendritic cell increasing agent (granule) 5 in an edible synthetic medium (with 0.5% yeast extract and 0.1% trypticase peptone), Lactobacillus helveticus strain SBT2171 After inoculating a weight% and culturing at 38 ° C. for 15 hours, the cells were collected by centrifugation. The collected cells were lyophilized to obtain a freeze-dried powder of the cells. 1 g of this lyophilized powder was mixed with 5 g of lactose and formed into granules to obtain the plasmacytoid dendritic cell increasing agent of the present invention.
  • Example 3 Production of plasmacytoid dendritic cell increasing agent (granule) Synthetic medium edible with Lactobacillus helveticus JCM-1120 strain (0.5% yeast extract, 0.1% trypticase peptone added) 5% by weight was inoculated and cultured at 38 ° C. for 15 hours, and the cells were collected by centrifugation. The collected cells were lyophilized to obtain a freeze-dried powder of the cells. 1 g of this lyophilized powder was mixed with 5 g of lactose and formed into granules to obtain the plasmacytoid dendritic cell increasing agent of the present invention.
  • Example 4 Manufacture of a plasmacytoid dendritic cell increasing agent (powder)
  • the Lactobacillus helveticus SBT2171 obtained in Example 2 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Description of Drugs General Rules "Powder” Lactose (JP) 400 g and potato starch (JP) 600 g were added to 10 g of the lyophilized powder of the strain and mixed uniformly to produce the plasmacytoid dendritic cell increasing agent of the present invention.
  • Example 5 Manufacture of a plasmacytoid dendritic cell increasing agent (powder)
  • the Lactobacillus helveticus JCM obtained in Example 3 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia explanation general formulation "powder" Lactose (JP) 400 g and potato starch (JP) 600 g were added to 10 g of lyophilized powder of the -1120 strain and mixed uniformly to produce the plasmacytoid dendritic cell increasing agent of the present invention.
  • Example 6 Production of nutritional health food for increasing plasma-like dendritic cells in the form of sticks 40 g of vitamin C or an equal mixture of vitamin C and citric acid, 100 g of granulated sugar, 60 g of an equal mixture of corn starch and lactose 40 g of lyophilized powder of Lactobacillus helveticus SBT2171 strain obtained in Example 2 was added and mixed. The mixture was packed in a bag to produce 150 bags of the stick-like nutritional health food of the present invention.
  • Example 7 Production of feed for increasing plasmacytoid dendritic cells Soybean meal 12 kg, skim milk powder 14 kg, soybean oil 4 kg, corn oil 2 kg, palm oil 23.2 kg, corn starch 14 kg, wheat flour 9 kg, bran 2 kg, vitamin mixture 5 kg, 2.8 kg of cellulose and 2 kg of a mineral mixture were blended, sterilized at 120 ° C. for 4 minutes, and 10 kg of Lactobacillus helveticus SBT2171 strain obtained in Example 2 was blended to produce a feed.
  • Lactobacillus helveticus SBT2171 The name and address of the depository institution where the biological material was deposited National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center Date 1-3-1 Higashi 1-chome, Tsukuba City, Ibaraki, Japan Date of deposit of biological material in Heisei June 22, 6 (June 22, 1994) (original deposit date) March 6, 1996 (March 6, 1996) (Date of transfer to deposit under the Budapest Treaty by original deposit) The deposit number assigned by the depository in Hai for the deposit FERM BP-5445

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Abstract

La présente invention aborde le problème de la fourniture d'un produit pharmaceutique, d'un aliment ou d'une boisson et d'un aliment pour animaux ayant un effet favorisant la prolifération des cellules dendritiques plasmocytoïdes, exerçant ainsi une double action de prévention des infections et de prévention des maladies inflammatoires. L'utilisation de bactéries d'acide lactique du genre Lactobacillus helveticus ou d'une culture de celles-ci en tant que principe actif permet de proposer un produit pharmaceutique, un aliment ou une boisson et un aliment pour animaux destinés à favoriser la prolifération des cellules dendritiques plasmacytoïdes à l'intérieur du corps.
PCT/JP2016/071814 2015-08-04 2016-07-26 Promoteur de prolifération de cellules dendritiques plasmacytoïdes WO2017022560A1 (fr)

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JP2015153961A JP6705628B2 (ja) 2015-08-04 2015-08-04 形質細胞様樹状細胞誘導剤
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019210242A (ja) * 2018-06-04 2019-12-12 雪印メグミルク株式会社 ラクトバチルス属の乳酸菌を有効成分とするｓｒｃａｐ発現抑制用組成物
JP2020080785A (ja) * 2018-11-29 2020-06-04 雪印メグミルク株式会社 歯周病予防用組成物

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JP2007099628A (ja) * 2005-09-30 2007-04-19 Snow Brand Milk Prod Co Ltd 免疫機能調節剤および免疫機能調節用飲食品
JP2011121923A (ja) * 2009-12-14 2011-06-23 Snow Brand Milk Prod Co Ltd 制御性ｔ細胞増加剤

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JP2007099628A (ja) * 2005-09-30 2007-04-19 Snow Brand Milk Prod Co Ltd 免疫機能調節剤および免疫機能調節用飲食品
JP2011121923A (ja) * 2009-12-14 2011-06-23 Snow Brand Milk Prod Co Ltd 制御性ｔ細胞増加剤

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GOUBIER ANNE ET AL.: "Plasmacytoid dendritic cells mediate oral tolerance", IMMUNITY, vol. 29, no. 3, 2008, pages 464 - 75, XP055361881 *
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019210242A (ja) * 2018-06-04 2019-12-12 雪印メグミルク株式会社 ラクトバチルス属の乳酸菌を有効成分とするｓｒｃａｐ発現抑制用組成物
JP7224114B2 (ja) 2018-06-04 2023-02-17 雪印メグミルク株式会社 ラクトバチルス属の乳酸菌を有効成分とするｓｒｃａｐ発現抑制用組成物
JP2020080785A (ja) * 2018-11-29 2020-06-04 雪印メグミルク株式会社 歯周病予防用組成物
JP7160652B2 (ja) 2018-11-29 2022-10-25 雪印メグミルク株式会社 歯周病予防用組成物
JP7410250B2 (ja) 2018-11-29 2024-01-09 雪印メグミルク株式会社 歯周病予防用組成物

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