WO2017057307A1 - Antibody production controller - Google Patents
Antibody production controller Download PDFInfo
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- WO2017057307A1 WO2017057307A1 PCT/JP2016/078337 JP2016078337W WO2017057307A1 WO 2017057307 A1 WO2017057307 A1 WO 2017057307A1 JP 2016078337 W JP2016078337 W JP 2016078337W WO 2017057307 A1 WO2017057307 A1 WO 2017057307A1
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- production
- antibody
- lactobacillus helveticus
- iga
- lactic acid
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- 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
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
-
- 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 an antibody production regulator, a food / beverage product, a nutritional composition, and a feed, which contains lactic acid bacteria belonging to Lactobacillus helveticus as an active ingredient.
- the immune system is an advanced biological defense mechanism that plays a role in eliminating pathogens such as bacteria and viruses that try to enter the body and tumor cells generated in the body.
- immunoglobulins antibodies having a function of recognizing and binding molecules (antigens) such as specific proteins play an important role.
- IgA antibody which is a kind of immunoglobulin, is the most abundantly produced antibody in the body and is secreted into mucous membranes such as intestinal tract, respiratory tract and oral cavity.
- IgA antibodies produced in the intestine have the function of blocking the attachment and infection of pathogenic microorganisms to intestinal epithelial cells and neutralizing bacterial toxins.
- IgA antibodies are also known to inhibit food allergies by binding to food-derived antigens that become allergens in the intestine and blocking passage through the intestinal wall. Therefore, it has been expected to provide drugs or food ingredients that promote the production of IgA antibodies in the intestine and prevent infections and food allergies.
- autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease
- the production of IgG antibodies which are autoantibodies whose antigens are constituents of the body, are caused by the breakdown of the immune system. It is supposed to be one.
- Rheumatoid arthritis is an autoimmune disease in which the immune system destroys joint tissue and is the most common autoimmune disease affecting approximately 1% of the population.
- IgG antibodies are the most common.
- Factors that promote the production of IgA antibodies and factors that inhibit the production of IgG antibodies, which are autoantibodies have been studied so far.
- Factors that promote the production of IgA antibodies include Bifidobacterium (Patent Document 1), Lactobacillus (Patent Document 2), and corn poppy extract (Patent Document 3). Lactobacillus spp.
- Patent Document 4 has been reported as a factor that suppresses the production of IgG antibodies that are autoantibodies. Further, according to Non-Patent Document 1, there are not only bacterial species that promote and suppress IgA antibody production depending on the type of lactic acid bacteria, but also strains that promote and inhibit IgA antibody production in the same bacterial species. There is. There is also a description that Lactobacillus helveticus was not selected as a lactic acid bacterium with high IgA production when screening a lactic acid bacterium with high IgA production from a plurality of lactic acid bacteria (Non-patent Document 1).
- Lactobacillus plantarum Strain AYA Enhances IgA Secretion and Provides Survival Protection against Influenza Virus Infection in Mice January 2014
- Patent Documents 1 to 4 describe factors that promote the production of IgA antibodies or antibody production regulators that suppress the production of IgG antibodies that are autoantibodies.
- An antibody production regulator having a combination of Therefore the present invention promotes the production of IgA antibody and suppresses the production of IgG antibody, which is an autoantibody, thereby providing a medicinal product, nutrition that can bring about the effects of both prevention of infectious diseases and prevention of autoimmune diseases. It aims at providing a composition, food-drinks, and feed.
- the present inventors have found that lactic acid bacteria belonging to Lactobacillus helvetivcus, In particular, Lactobacillus helvetivcus SBT2171 strain was found and the present invention was completed. That is, the present invention has the following configuration. (1) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or a culture thereof as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies.
- IgA antibody according to (1) wherein the lactic acid bacterium belonging to Lactobacillus helveticus is Lactobacillus helveticus SBT2171 (FERM BP-5445)
- An antibody production regulator that promotes and suppresses the production of IgG antibodies that are autoantibodies.
- An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use method of antibody production regulator to administer.
- An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies
- the present invention enables prevention and improvement of an excessive autoimmune response as well as infection prevention effects against viruses and bacteria. Furthermore, when a human or the like eats a cheese or lactic acid bacteria beverage containing a high content of the lactic acid bacteria of the present invention, it is possible to regulate antibody production.
- Lactobacillus helveticus As the lactic acid bacterium of the present invention, lactic acid bacteria belonging to Lactobacillus helveticus can be used.
- Lactobacillus helveticus SBT2171 strain (deposited as FERM BP-5445), SBT-2161 strain (deposited as NITE BP-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.
- Bacteria isolated from the obtained culture by means of collection such as centrifugation can be used as it is as an active ingredient of the present invention.
- the cells can be concentrated, dried, freeze-dried, etc., or may be killed by heat drying. Not only those that have been purely isolated as bacterial cells, but also cultures, suspensions, other bacterial cell-containing materials, and cytoplasm and cell wall fractions obtained by treating bacterial cells with enzymes or physical means can be used in the present invention. It can be used as an active ingredient.
- the form of the culture is not only a culture using a medium generally used for cultivation of lactic acid bacteria, such as MRS medium (DIFCO) which is a synthetic medium, reduced skim milk medium, cheese, fermented milk, dairy products
- MRS medium DIFCO
- dairy products such as lactic acid bacteria beverages are not particularly limited.
- 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.
- Formulation may be carried out by appropriately mixing excipients, stabilizers, flavoring agents, etc. that are permitted in the formulation, concentrating and freeze-drying, or by heating to dry cells.
- the active ingredient of the present invention includes these dried products, concentrates, and pastes.
- excipients, binders, disintegrants, lubricants, flavoring agents, suspending agents, coating agents, and other optional 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 antibody production regulator 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 antibody production regulator of the present invention can be added to feed. Like the said food-drinks, you may mix
- Lactobacillus helveticus (Lactobacillus helveticus) is used in combination with antibody production regulators, antibody production regulation foods and drinks, nutritional compositions, feeds, etc., or processed products of these materials
- Lactobacillus helveticus (Lactobacillus helveticus)
- the mixing ratio of helveticus) is not particularly limited, and may be appropriately adjusted according to the ease of production and the preferred daily dose. It is determined individually in consideration of the symptom, age, etc. of the administration subject, but in the case of normal adults, 10 to 200 g of Lactobacillus helveticus culture etc. What is necessary is just to adjust compounding quantity etc. so that 5,000mg can be ingested. A desired effect can be exhibited by ingesting in this way.
- the tissue lysate was centrifuged at 10,000 ⁇ g for 10 minutes, and the supernatant was collected as a tissue extract and stored at ⁇ 80 ° C.
- the amount of IgA antibody in the tissue extract was measured using an ELISA kit (Bethyl Laboratories).
- the protein concentration in the tissue extract was measured using a BCA kit (Thermo Scientific). The value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG.
- Bovine type II collagen (CII) (2 mg / ml in 0.05M acetic acid, Chondrex) and Complete Freund's Adjuvant (CFA, Chondrex) were combined in equal amounts to produce a 1 mg / ml emulsion as collagen for immunization. This was intradermally administered at two locations (total 100 ⁇ g / mouse) of 50 ⁇ L at the ridge of all mice.
- the second collagen administration was performed 21 days after the first collagen administration day in the same manner as the first administration. From the second collagen administration day, they were raised for 3 weeks and dissected.
- SBT217 cells suspended in 0.35M NaHCO 3 / PBS at a concentration of 100 mg / mL were administered daily from the first collagen administration day to the day before dissection. It was orally administered so that the body weight was 30 mg / mouse.
- 0.35M NaHCO 3 / PBS was orally administered so as to be 300 ⁇ L / mouse. NaHCO 3 was added for the purpose of neutralizing gastric acid. Blood collected at the time of dissection was centrifuged to separate serum, and stored at -80 ° C. The amount of CII-specific IgG antibody in the serum was measured using an ELISA kit (Chondrex). The amount of IgG antibody is shown in FIG.
- 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 SBT2171 strain (FERM BP-5445) were added, and 0.003% of rennet was further added. Solidified.
- the curd thus obtained was cut, stirred until the pH was 6.2 to 6.1, and whey was discharged to obtain curd grains. Then, the curd grains were filled and pressed, further salted, and aged at 10 ° C.
- This hard natural cheese (aged for 6 months) was minced using a mincer (GM-DX, manufactured by Nippon Carrier Co., Ltd.) and freeze-dried. Then, it refined
- the cheese powder can be used as it is as an antibody production regulator of the present invention.
- Test Example 3 Oral administration test of Lactobacillus helveticus SBT2171 culture to mice (IgA antibody level) 1. 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 IgA antibody amount and protein concentration after dissection were measured, and the value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG. 2. Test results From FIG.
- the amount of protein in the group (SBT2171 group) ingesting the diet supplemented with the culture of Lactobacillus helveticus SBT2171 strain was greater than that in the group ingesting the diet not added (control group) It was found that the amount of IgA production per unit was high. That is, it was shown that the production of IgA antibody is promoted by ingestion of a culture of Lactobacillus helveticus.
- Test Example 4 Oral administration test of Lactobacillus helveticus SBT2171 strain culture to mice (IgG antibody level) 1. 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 2. The amount of CII-specific IgG antibody in the serum after dissection was measured, and the results are shown in FIG. 2. Test results From FIG. 4, the group that ingested the diet supplemented with the culture of Lactobacillus helveticus SBT2171 (SBT2171 group) was more CII-specific than the group that ingested the diet not added (control group) It was found that the production amount of IgG antibody was low. That is, it was shown that the ingestion of a culture of Lactobacillus helveticus suppresses the production of IgG antibodies that are autoantibodies.
- Example 2 Production of antibody production regulator (granule) Lactobacillus helveticus SBT2171 strain was inoculated into 5% by weight in an edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added). After 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 antibody production regulator of the present invention.
- Example 3 Manufacture of antibody production regulator (granule) 5% by weight of Lactobacillus helveticus JCM-1120 strain in edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added) After inoculation 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 antibody production regulator of the present invention.
- Example 4 Manufacture of antibody production regulator (powder)
- the Lactobacillus helveticus SBT2171 obtained in Example 2 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder” Lactose (JP) 40 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 antibody production regulator of the present invention.
- Example 5 Manufacture of antibody production regulator (powder)
- the Lactobacillus helveticus JCM obtained in Example 3 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder” Lactose (JP) 400 g and potato starch (JP) 600 g were added to 10 g of freeze-dried powder of strain -1120 and mixed uniformly to produce the antibody production regulator of the present invention.
- Example 6 Manufacture of stick-like nutritional health foods Vitamin C 40 g or an equivalent mixture of vitamin C and citric acid 40 g, granulated sugar 100 g, corn starch and lactose equivalent mixture 60 g obtained in Example 2 above 40 g of the lyophilized powder of the obtained Lactobacillus helveticus SBT2171 strain 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 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 mineral mixture were 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. .
- an excessive autoimmune response can be prevented and improved together with an infection preventing effect against viruses and bacteria. Furthermore, it is also possible to regulate antibody production by eating cheese or lactic acid bacteria beverages that are anti-containing the lactic acid bacteria.
- 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
Abstract
The present invention addresses the problem of providing a drug, a nutritional composition, a food, a drink and a feed, each being capable of promoting the production of IgA antibody and suppressing the production of IgG antibody, which is an autoantibody, so as to achieve both an effect of preventing an infectious disease and an effect of preventing an autoimmune disease. The present inventors conducted intensive studies to search for a factor promoting the production of IgA antibody and suppressing the production of IgG antibody, which is an autoantibody, in vivo and consequently found a lactic acid bacterium of Lactobacillus helveticus, thereby completing the present invention. The constitution of the present invention is as follows. (1) An antibody production controller promoting the production of IgA antibody and suppressing the production of IgG antibody which is an autoantibody, said antibody production controller comprising, as an active ingredient, a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof.
Description
本発明は、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌を有効成分とする、抗体産生調節剤、飲食品、栄養組成物および飼料に関する。
The present invention relates to an antibody production regulator, a food / beverage product, a nutritional composition, and a feed, which contains lactic acid bacteria belonging to Lactobacillus helveticus as an active ingredient.
免疫系は、体内に侵入しようとする細菌やウイルスなどの病原体や体内で生じた腫瘍細胞を排除する役割を担う高度な生体防御機構である。免疫系では、特定のタンパク質などの分子（抗原）を認識して結合する働きをもつ免疫グロブリン（抗体）が重要な役割を果たしている。免疫グロブリンの一種であるIgA抗体は、体内で最も多量に産生される抗体であり、腸管、気道および口腔などの粘膜に分泌されている。腸内で産生されるIgA抗体は、病原性微生物の腸管上皮細胞への付着および感染を阻止し、細菌由来毒素を中和する機能がある。また、IgA抗体は、腸内においてアレルゲンとなる食品由来の抗原と結合し、腸管壁の通過を阻止することで、食物アレルギーを抑制することも知られている。そのため、腸内でIgA抗体の産生を促進し、感染症と食物アレルギーを予防するような薬剤または食品成分の提供が期待されてきた。
一方、関節リウマチおよび炎症性腸疾患などの自己免疫疾患においては、免疫系の破綻によって自己の体の構成成分を抗原とする自己抗体であるIgG抗体が産生されることが、発症の原因の1つであるとされている。関節リウマチは、免疫系が関節の組織を破壊してしまう自己免疫疾患であり、人口の約１％が罹患する最も頻度の高い自己免疫疾患である。関節リウマチの患者の血液中には、様々な自己抗体が認められるが、IgG抗体が最も多い。そこで、生体内で自己抗体であるIgG抗体の産生を抑制し、自己免疫疾患を予防、緩和するような薬剤または食品成分の提供が期待されてきた。
IgA抗体の産生を促進する因子および自己抗体であるIgG抗体の産生を抑制する因子はこれまでにも研究されてきた。IgA抗体の産生を促進する因子としては、ビフィドバクテリム属（Bifidobacterium）菌（特許文献1）、ラクトバチルス属（Lactobacillus）菌（特許文献2）、ヒナゲシ抽出物（特許文献3）がある。自己抗体であるIgG抗体の産生を抑制する因子としては、ラクトバチルス属菌（特許文献4）が報告されている。また、非特許文献１によれば、乳酸菌の種類によってIgA抗体の産生を促進する菌種と抑制する菌種があるだけでなく、同一菌種においてもIgA抗体を産生促進する株と抑制する株がある。また、複数の乳酸菌からIgA産生が多い乳酸菌をスクリーニングする際に、IgA産生が多い乳酸菌としてラクトバチルス・ヘルベティカスが選ばれなかったという記載もある（非特許文献１）。 The immune system is an advanced biological defense mechanism that plays a role in eliminating pathogens such as bacteria and viruses that try to enter the body and tumor cells generated in the body. In the immune system, immunoglobulins (antibodies) having a function of recognizing and binding molecules (antigens) such as specific proteins play an important role. IgA antibody, which is a kind of immunoglobulin, is the most abundantly produced antibody in the body and is secreted into mucous membranes such as intestinal tract, respiratory tract and oral cavity. IgA antibodies produced in the intestine have the function of blocking the attachment and infection of pathogenic microorganisms to intestinal epithelial cells and neutralizing bacterial toxins. IgA antibodies are also known to inhibit food allergies by binding to food-derived antigens that become allergens in the intestine and blocking passage through the intestinal wall. Therefore, it has been expected to provide drugs or food ingredients that promote the production of IgA antibodies in the intestine and prevent infections and food allergies.
On the other hand, in autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease, the production of IgG antibodies, which are autoantibodies whose antigens are constituents of the body, are caused by the breakdown of the immune system. It is supposed to be one. Rheumatoid arthritis is an autoimmune disease in which the immune system destroys joint tissue and is the most common autoimmune disease affecting approximately 1% of the population. Various autoantibodies are found in the blood of patients with rheumatoid arthritis, but IgG antibodies are the most common. Thus, it has been expected to provide drugs or food ingredients that suppress the production of IgG antibodies, which are autoantibodies in vivo, and prevent or alleviate autoimmune diseases.
Factors that promote the production of IgA antibodies and factors that inhibit the production of IgG antibodies, which are autoantibodies, have been studied so far. Factors that promote the production of IgA antibodies include Bifidobacterium (Patent Document 1), Lactobacillus (Patent Document 2), and corn poppy extract (Patent Document 3). Lactobacillus spp. (Patent Document 4) has been reported as a factor that suppresses the production of IgG antibodies that are autoantibodies. Further, according to Non-Patent Document 1, there are not only bacterial species that promote and suppress IgA antibody production depending on the type of lactic acid bacteria, but also strains that promote and inhibit IgA antibody production in the same bacterial species. There is. There is also a description that Lactobacillus helveticus was not selected as a lactic acid bacterium with high IgA production when screening a lactic acid bacterium with high IgA production from a plurality of lactic acid bacteria (Non-patent Document 1).
一方、関節リウマチおよび炎症性腸疾患などの自己免疫疾患においては、免疫系の破綻によって自己の体の構成成分を抗原とする自己抗体であるIgG抗体が産生されることが、発症の原因の1つであるとされている。関節リウマチは、免疫系が関節の組織を破壊してしまう自己免疫疾患であり、人口の約１％が罹患する最も頻度の高い自己免疫疾患である。関節リウマチの患者の血液中には、様々な自己抗体が認められるが、IgG抗体が最も多い。そこで、生体内で自己抗体であるIgG抗体の産生を抑制し、自己免疫疾患を予防、緩和するような薬剤または食品成分の提供が期待されてきた。
IgA抗体の産生を促進する因子および自己抗体であるIgG抗体の産生を抑制する因子はこれまでにも研究されてきた。IgA抗体の産生を促進する因子としては、ビフィドバクテリム属（Bifidobacterium）菌（特許文献1）、ラクトバチルス属（Lactobacillus）菌（特許文献2）、ヒナゲシ抽出物（特許文献3）がある。自己抗体であるIgG抗体の産生を抑制する因子としては、ラクトバチルス属菌（特許文献4）が報告されている。また、非特許文献１によれば、乳酸菌の種類によってIgA抗体の産生を促進する菌種と抑制する菌種があるだけでなく、同一菌種においてもIgA抗体を産生促進する株と抑制する株がある。また、複数の乳酸菌からIgA産生が多い乳酸菌をスクリーニングする際に、IgA産生が多い乳酸菌としてラクトバチルス・ヘルベティカスが選ばれなかったという記載もある（非特許文献１）。 The immune system is an advanced biological defense mechanism that plays a role in eliminating pathogens such as bacteria and viruses that try to enter the body and tumor cells generated in the body. In the immune system, immunoglobulins (antibodies) having a function of recognizing and binding molecules (antigens) such as specific proteins play an important role. IgA antibody, which is a kind of immunoglobulin, is the most abundantly produced antibody in the body and is secreted into mucous membranes such as intestinal tract, respiratory tract and oral cavity. IgA antibodies produced in the intestine have the function of blocking the attachment and infection of pathogenic microorganisms to intestinal epithelial cells and neutralizing bacterial toxins. IgA antibodies are also known to inhibit food allergies by binding to food-derived antigens that become allergens in the intestine and blocking passage through the intestinal wall. Therefore, it has been expected to provide drugs or food ingredients that promote the production of IgA antibodies in the intestine and prevent infections and food allergies.
On the other hand, in autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease, the production of IgG antibodies, which are autoantibodies whose antigens are constituents of the body, are caused by the breakdown of the immune system. It is supposed to be one. Rheumatoid arthritis is an autoimmune disease in which the immune system destroys joint tissue and is the most common autoimmune disease affecting approximately 1% of the population. Various autoantibodies are found in the blood of patients with rheumatoid arthritis, but IgG antibodies are the most common. Thus, it has been expected to provide drugs or food ingredients that suppress the production of IgG antibodies, which are autoantibodies in vivo, and prevent or alleviate autoimmune diseases.
Factors that promote the production of IgA antibodies and factors that inhibit the production of IgG antibodies, which are autoantibodies, have been studied so far. Factors that promote the production of IgA antibodies include Bifidobacterium (Patent Document 1), Lactobacillus (Patent Document 2), and corn poppy extract (Patent Document 3). Lactobacillus spp. (Patent Document 4) has been reported as a factor that suppresses the production of IgG antibodies that are autoantibodies. Further, according to Non-Patent Document 1, there are not only bacterial species that promote and suppress IgA antibody production depending on the type of lactic acid bacteria, but also strains that promote and inhibit IgA antibody production in the same bacterial species. There is. There is also a description that Lactobacillus helveticus was not selected as a lactic acid bacterium with high IgA production when screening a lactic acid bacterium with high IgA production from a plurality of lactic acid bacteria (Non-patent Document 1).
特許文献１～４にはIgA抗体の産生を促進する因子または自己抗体であるIgG抗体の産生を抑制する抗体産生調節因子が記載されている。しかし、日常的に継続摂取することで、感染症と自己免疫疾患の両方を予防するためには、単一の因子でIgA抗体産生の促進と自己抗体であるIgG抗体産生の抑制の両方の作用を合わせもつような抗体産生調節因子が望ましい。
そこで、本発明は、IgA抗体の産生を促進し、自己抗体であるIgG抗体の産生を抑制することで、感染症の予防と自己免疫疾患の予防の両者の効果をもたらすことができる医薬品、栄養組成物、飲食品および飼料を提供することを目的とする。 Patent Documents 1 to 4 describe factors that promote the production of IgA antibodies or antibody production regulators that suppress the production of IgG antibodies that are autoantibodies. However, in order to prevent both infectious diseases and autoimmune diseases by continuous daily intake, both the effects of promoting the production of IgA antibodies and suppressing the production of IgG antibodies, which are autoantibodies, can be prevented. An antibody production regulator having a combination of
Therefore, the present invention promotes the production of IgA antibody and suppresses the production of IgG antibody, which is an autoantibody, thereby providing a medicinal product, nutrition that can bring about the effects of both prevention of infectious diseases and prevention of autoimmune diseases. It aims at providing a composition, food-drinks, and feed.
そこで、本発明は、IgA抗体の産生を促進し、自己抗体であるIgG抗体の産生を抑制することで、感染症の予防と自己免疫疾患の予防の両者の効果をもたらすことができる医薬品、栄養組成物、飲食品および飼料を提供することを目的とする。 Patent Documents 1 to 4 describe factors that promote the production of IgA antibodies or antibody production regulators that suppress the production of IgG antibodies that are autoantibodies. However, in order to prevent both infectious diseases and autoimmune diseases by continuous daily intake, both the effects of promoting the production of IgA antibodies and suppressing the production of IgG antibodies, which are autoantibodies, can be prevented. An antibody production regulator having a combination of
Therefore, the present invention promotes the production of IgA antibody and suppresses the production of IgG antibody, which is an autoantibody, thereby providing a medicinal product, nutrition that can bring about the effects of both prevention of infectious diseases and prevention of autoimmune diseases. It aims at providing a composition, food-drinks, and feed.
本発明者らは、生体内で、IgA抗体の産生を促進し、自己抗体であるIgG抗体の産生を抑制する因子を鋭意探索した結果、乳酸菌のラクトバチルス・ヘルベティカス（Lactobacillus helvetivcus）に属する乳酸菌、特にラクトバチルス・ヘルベティカス（Lactobacillus helvetivcus）SBT2171株を見出し、本発明を完成させるに至った。
即ち本発明は以下の構成を有する。
（１）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。
（２）前記ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌がラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171（FERM BP-5445）であることを特徴とする、（１）に記載の IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。
（３）（１）または（２）に記載の剤を添加したIgG抗体の産生を抑制する抗体産生調節用栄養組成物、飲食品又は飼料。
（４）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する抗体産生調節剤の使用方法。
（５）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する感染症の予防と自己免疫疾患の予防のための抗体産生調節剤の使用方法。
（６）IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤の製造における、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物の使用。 As a result of intensive search for factors that promote the production of IgA antibodies and suppress the production of IgG antibodies that are autoantibodies in vivo, the present inventors have found that lactic acid bacteria belonging to Lactobacillus helvetivcus, In particular, Lactobacillus helvetivcus SBT2171 strain was found and the present invention was completed.
That is, the present invention has the following configuration.
(1) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or a culture thereof as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies.
(2) The production of IgA antibody according to (1), wherein the lactic acid bacterium belonging to Lactobacillus helveticus is Lactobacillus helveticus SBT2171 (FERM BP-5445) An antibody production regulator that promotes and suppresses the production of IgG antibodies that are autoantibodies.
(3) A nutritional composition for regulating antibody production, food or drink, or feed that suppresses the production of IgG antibodies to which the agent according to (1) or (2) is added.
(4) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use method of antibody production regulator to administer.
(5) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use of an antibody production regulator for the prevention of infectious diseases and autoimmune diseases to be administered.
(6) Use of a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof in the production of an antibody production regulator that promotes production of IgA antibody and suppresses production of IgG antibody that is an autoantibody.
即ち本発明は以下の構成を有する。
（１）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。
（２）前記ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌がラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171（FERM BP-5445）であることを特徴とする、（１）に記載の IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。
（３）（１）または（２）に記載の剤を添加したIgG抗体の産生を抑制する抗体産生調節用栄養組成物、飲食品又は飼料。
（４）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する抗体産生調節剤の使用方法。
（５）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する感染症の予防と自己免疫疾患の予防のための抗体産生調節剤の使用方法。
（６）IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤の製造における、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物の使用。 As a result of intensive search for factors that promote the production of IgA antibodies and suppress the production of IgG antibodies that are autoantibodies in vivo, the present inventors have found that lactic acid bacteria belonging to Lactobacillus helvetivcus, In particular, Lactobacillus helvetivcus SBT2171 strain was found and the present invention was completed.
That is, the present invention has the following configuration.
(1) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or a culture thereof as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies.
(2) The production of IgA antibody according to (1), wherein the lactic acid bacterium belonging to Lactobacillus helveticus is Lactobacillus helveticus SBT2171 (FERM BP-5445) An antibody production regulator that promotes and suppresses the production of IgG antibodies that are autoantibodies.
(3) A nutritional composition for regulating antibody production, food or drink, or feed that suppresses the production of IgG antibodies to which the agent according to (1) or (2) is added.
(4) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use method of antibody production regulator to administer.
(5) An antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use of an antibody production regulator for the prevention of infectious diseases and autoimmune diseases to be administered.
(6) Use of a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof in the production of an antibody production regulator that promotes production of IgA antibody and suppresses production of IgG antibody that is an autoantibody.
本発明は、ウイルスや細菌などに対する感染予防効果とともに、過剰な自己免疫応答の予防、改善を可能とする。さらに、ヒト等が本発明の該乳酸菌を高含有するチーズまたは乳酸菌飲料を食することで、抗体産生の調節も可能となる。
The present invention enables prevention and improvement of an excessive autoimmune response as well as infection prevention effects against viruses and bacteria. Furthermore, when a human or the like eats a cheese or lactic acid bacteria beverage containing a high content of the lactic acid bacteria of the present invention, it is possible to regulate antibody production.
本発明の乳酸菌は、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌を用いることができる。特に、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株（FERM BP-5445として寄託）、SBT-2161株(NITE BP-01707として寄託)、SBT2195株（FERM P-11538として寄託）、SBT2196株（FERM P-11676として寄託）、SBT0064株（FERM P-21079として寄託）、SBT0402株（FERM P-21559として寄託）等が好ましいが、これらに限定されるものではない。
ラクトバチルス・ヘルベティカスは、乳酸菌培養の常法に従って培養することができる。培養培地は、乳培地又は乳成分を含む培地、これを含まない半合成培地など種々の培地を用いることができる。このような培地としては、還元脱脂乳培地などを例示することができる。得られた培養物から遠心分離などの集菌手段によって分離された菌体はそのまま本発明の有効成分として用いることができる。この菌体は濃縮、乾燥、凍結乾燥などした菌体を用いることもできるし、加熱乾燥などにより死菌体にしてもよい。
菌体として純粋に分離されたものだけでなく、培養物、懸濁物、その他の菌体含有物や、菌体を酵素や物理的手段を用いて処理した細胞質や細胞壁画分も本発明の有効成分として用いることができる。培養物などの形態は、合成培地であるMRS培地（DIFCO社製）、還元脱脂乳培地など一般的に乳酸菌の培養に用いられる培地を用いた培養物だけでなく、チーズ、発酵乳、乳製品乳酸菌飲料などの乳製品などを例示することができるが特に限定されるものではない。
さらに、得られた培養物から遠心分離、濾過操作などの方法を用いて、乳タンパク質沈殿や菌体成分を除去することによって調製した培養上清なども用いることができる。固形分が少ない上清であるため、飲食品などへの適用範囲が広くなる。例えば、還元脱脂乳培養物を5,000 rpm、10分間遠心分離することにより培養上清を調製することができる。 As the lactic acid bacterium of the present invention, lactic acid bacteria belonging to Lactobacillus helveticus can be used. In particular, Lactobacillus helveticus SBT2171 strain (deposited as FERM BP-5445), SBT-2161 strain (deposited as NITE BP-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. As the culture medium, 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. Bacteria isolated from the obtained culture by means of collection such as centrifugation can be used as it is as an active ingredient of the present invention. The cells can be concentrated, dried, freeze-dried, etc., or may be killed by heat drying.
Not only those that have been purely isolated as bacterial cells, but also cultures, suspensions, other bacterial cell-containing materials, and cytoplasm and cell wall fractions obtained by treating bacterial cells with enzymes or physical means can be used in the present invention. It can be used as an active ingredient. The form of the culture is not only a culture using a medium generally used for cultivation of lactic acid bacteria, such as MRS medium (DIFCO) which is a synthetic medium, reduced skim milk medium, cheese, fermented milk, dairy products Examples of dairy products such as lactic acid bacteria beverages are not particularly limited.
Furthermore, 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. For example, the culture supernatant can be prepared by centrifuging the reduced skim milk culture at 5,000 rpm for 10 minutes.
ラクトバチルス・ヘルベティカスは、乳酸菌培養の常法に従って培養することができる。培養培地は、乳培地又は乳成分を含む培地、これを含まない半合成培地など種々の培地を用いることができる。このような培地としては、還元脱脂乳培地などを例示することができる。得られた培養物から遠心分離などの集菌手段によって分離された菌体はそのまま本発明の有効成分として用いることができる。この菌体は濃縮、乾燥、凍結乾燥などした菌体を用いることもできるし、加熱乾燥などにより死菌体にしてもよい。
菌体として純粋に分離されたものだけでなく、培養物、懸濁物、その他の菌体含有物や、菌体を酵素や物理的手段を用いて処理した細胞質や細胞壁画分も本発明の有効成分として用いることができる。培養物などの形態は、合成培地であるMRS培地（DIFCO社製）、還元脱脂乳培地など一般的に乳酸菌の培養に用いられる培地を用いた培養物だけでなく、チーズ、発酵乳、乳製品乳酸菌飲料などの乳製品などを例示することができるが特に限定されるものではない。
さらに、得られた培養物から遠心分離、濾過操作などの方法を用いて、乳タンパク質沈殿や菌体成分を除去することによって調製した培養上清なども用いることができる。固形分が少ない上清であるため、飲食品などへの適用範囲が広くなる。例えば、還元脱脂乳培養物を5,000 rpm、10分間遠心分離することにより培養上清を調製することができる。 As the lactic acid bacterium of the present invention, lactic acid bacteria belonging to Lactobacillus helveticus can be used. In particular, Lactobacillus helveticus SBT2171 strain (deposited as FERM BP-5445), SBT-2161 strain (deposited as NITE BP-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. As the culture medium, 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. Bacteria isolated from the obtained culture by means of collection such as centrifugation can be used as it is as an active ingredient of the present invention. The cells can be concentrated, dried, freeze-dried, etc., or may be killed by heat drying.
Not only those that have been purely isolated as bacterial cells, but also cultures, suspensions, other bacterial cell-containing materials, and cytoplasm and cell wall fractions obtained by treating bacterial cells with enzymes or physical means can be used in the present invention. It can be used as an active ingredient. The form of the culture is not only a culture using a medium generally used for cultivation of lactic acid bacteria, such as MRS medium (DIFCO) which is a synthetic medium, reduced skim milk medium, cheese, fermented milk, dairy products Examples of dairy products such as lactic acid bacteria beverages are not particularly limited.
Furthermore, 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. For example, the culture supernatant can be prepared by centrifuging the reduced skim milk culture at 5,000 rpm for 10 minutes.
製剤化は製剤上許可されている賦型剤、安定剤、矯味剤などを適宜混合して濃縮、凍結乾燥するほか、加熱乾燥して死菌体にすることで行ってもよい。本発明の有効成分には、これらの乾燥物、濃縮物、ペースト状物も含有される。また、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の抗体産生調節作用を妨げない範囲で、賦型剤、結合剤、崩壊剤、滑沢剤、矯味矯臭剤、懸濁剤、コーティング剤、その他の任意の薬剤を混合して製剤化することもできる。剤形は、錠剤、カプセル剤、顆粒剤、散剤、粉剤、シロップ剤などが可能であり、これらを経口的に投与することが望ましい。
Formulation may be carried out by appropriately mixing excipients, stabilizers, flavoring agents, etc. that are permitted in the formulation, concentrating and freeze-drying, or by heating to dry cells. The active ingredient of the present invention includes these dried products, concentrates, and pastes. In addition, as long as it does not interfere with the antibody production control effect of Lactobacillus helveticus, excipients, binders, disintegrants, lubricants, flavoring agents, suspending agents, coating agents, and other optional 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.
本発明の抗体産生調節剤はどのような飲食品に配合しても良く、飲食品の製造工程中に原料に添加しても良い。飲食品の例としては、チーズ、発酵乳、乳製品乳酸菌飲料、乳酸菌飲料、バター、マーガリンなどの乳製品、乳飲料、果汁飲料、清涼飲料などの飲料、ゼリー、キャンディー、プリン、マヨネーズなどの卵加工品、バターケーキなどの菓子・パン類、さらには、各種粉乳の他、乳幼児食品、栄養組成物などを挙げることができるが特に限定されるものではない。
The antibody production regulator 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. Examples of 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.
さらに、本発明の抗体産生調節剤は飼料に配合することができる。前記飲食品と同様にどのような飼料に配合しても良く、飼料の製造工程中に原料に添加しても良い。
Furthermore, the antibody production regulator of the present invention can be added to feed. Like the said food-drinks, you may mix | blend with what kind of feed, and may add to a raw material during the manufacturing process of a feed.
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）を抗体産生調節剤あるいは、抗体産生調節用飲食品、栄養組成物、飼料などの素材又はそれら素材の加工品に配合させて使用する場合、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の配合割合は特に限定されず、製造の容易性や好ましい一日投与量にあわせて適宜調節すればよい。投与対象者の症状、年齢などを考慮してそれぞれ個別に決定されるが、通常成人の場合、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の培養物などを10～200ｇ、あるいはその菌体自体を0.1～5,000 mg摂取できるように配合量などを調整すればよい。このようにして摂取することにより所望の効果を発揮することができる。
When Lactobacillus helveticus (Lactobacillus helveticus) is used in combination with antibody production regulators, antibody production regulation foods and drinks, nutritional compositions, feeds, etc., or processed products of these materials, Lactobacillus helveticus (Lactobacillus helveticus) The mixing ratio of helveticus) is not particularly limited, and may be appropriately adjusted according to the ease of production and the preferred daily dose. It is determined individually in consideration of the symptom, age, etc. of the administration subject, but in the case of normal adults, 10 to 200 g of Lactobacillus helveticus culture etc. What is necessary is just to adjust compounding quantity etc. so that 5,000mg can be ingested. A desired effect can be exhibited by ingesting in this way.
以下に、実施例及び試験例を示し、本発明についてより詳細に説明するが、これらは単に例示するのみであり、本発明はこれらによって何ら限定されるものではない。
Hereinafter, the present invention will be described in more detail with reference to examples and test examples. However, these are merely illustrative, and the present invention is not limited to these examples.
［試験例１］
Balb/cマウスへの生菌ラクトバチルス・ヘルベティカス（Lactobacillus helveticus） SBT2171株菌体の経口投与試験（IgA抗体量評価）
１－１．試験方法
24匹のBalb/cマウス（7週齢、雄性）を、コントロール（contorol）群とSBT2171群の2群に分けた（n=12）。コントロール群には標準飼料（AIN-93G）を、SBT2171群には標準飼料（AIN-93G）にラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末を0.5％添加した飼料を自由に摂取させた。摂取開始から5週間後に解剖して小腸を摘出した。小腸から腸間膜脂肪を除去した後、生理食塩水で内容物を洗浄した。小腸の胃と接する側の1cmを除去し、除去部位から小腸を5 cm切り出して、液体窒素で凍結した後に-80℃で保存した。
小腸組織に40倍量の抽出バッファー（1×PBS、50 mM Tris-HCl（pH 6.8）、Protease inhibitor cocktail（Roche cOmplete））を加え、ホモジェナイザーを用いて氷中で破砕し組織破砕液を得た。組織破砕液を10,000×gで10分間遠心した後、上清を組織抽出液として回収し、-80℃で保存した。
組織抽出液中のIgA抗体量を、ELISAキット（Bethyl Laboratories）を用いて測定した。同時に、組織抽出液中のタンパク質濃度を、BCAキット（Thermo Scientific）を用いて測定した。IgA抗体量をタンパク質濃度で割った値を図１に示した。 [Test Example 1]
Oral administration test of Lactobacillus helveticus SBT2171 cells to Balb / c mice (Evaluation of IgA antibody amount)
1-1. Test Method Twenty-four Balb / c mice (7 weeks old, male) were divided into two groups, a control group and an SBT2171 group (n = 12). Standard feed (AIN-93G) was given to the control group, and the standard feed (AIN-93G) to the SBT2171 group was freely fed with 0.5% lyophilized powder of Lactobacillus helveticus SBT2171 strain. It was. After 5 weeks from the start of ingestion, the small intestine was removed by dissection. After removing mesenteric fat from the small intestine, the contents were washed with physiological saline. 1 cm on the side of the small intestine in contact with the stomach was removed, 5 cm of the small intestine was excised from the removed site, frozen in liquid nitrogen, and stored at −80 ° C.
Add 40 times the amount of extraction buffer (1 × PBS, 50 mM Tris-HCl (pH 6.8), Protease inhibitor cocktail (Roche cOmplete)) to the small intestine tissue, crush it in ice using a homogenizer, and remove the tissue lysate. Obtained. The tissue lysate was centrifuged at 10,000 × g for 10 minutes, and the supernatant was collected as a tissue extract and stored at −80 ° C.
The amount of IgA antibody in the tissue extract was measured using an ELISA kit (Bethyl Laboratories). At the same time, the protein concentration in the tissue extract was measured using a BCA kit (Thermo Scientific). The value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG.
Balb/cマウスへの生菌ラクトバチルス・ヘルベティカス（Lactobacillus helveticus） SBT2171株菌体の経口投与試験（IgA抗体量評価）
１－１．試験方法
24匹のBalb/cマウス（7週齢、雄性）を、コントロール（contorol）群とSBT2171群の2群に分けた（n=12）。コントロール群には標準飼料（AIN-93G）を、SBT2171群には標準飼料（AIN-93G）にラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末を0.5％添加した飼料を自由に摂取させた。摂取開始から5週間後に解剖して小腸を摘出した。小腸から腸間膜脂肪を除去した後、生理食塩水で内容物を洗浄した。小腸の胃と接する側の1cmを除去し、除去部位から小腸を5 cm切り出して、液体窒素で凍結した後に-80℃で保存した。
小腸組織に40倍量の抽出バッファー（1×PBS、50 mM Tris-HCl（pH 6.8）、Protease inhibitor cocktail（Roche cOmplete））を加え、ホモジェナイザーを用いて氷中で破砕し組織破砕液を得た。組織破砕液を10,000×gで10分間遠心した後、上清を組織抽出液として回収し、-80℃で保存した。
組織抽出液中のIgA抗体量を、ELISAキット（Bethyl Laboratories）を用いて測定した。同時に、組織抽出液中のタンパク質濃度を、BCAキット（Thermo Scientific）を用いて測定した。IgA抗体量をタンパク質濃度で割った値を図１に示した。 [Test Example 1]
Oral administration test of Lactobacillus helveticus SBT2171 cells to Balb / c mice (Evaluation of IgA antibody amount)
1-1. Test Method Twenty-four Balb / c mice (7 weeks old, male) were divided into two groups, a control group and an SBT2171 group (n = 12). Standard feed (AIN-93G) was given to the control group, and the standard feed (AIN-93G) to the SBT2171 group was freely fed with 0.5% lyophilized powder of Lactobacillus helveticus SBT2171 strain. It was. After 5 weeks from the start of ingestion, the small intestine was removed by dissection. After removing mesenteric fat from the small intestine, the contents were washed with physiological saline. 1 cm on the side of the small intestine in contact with the stomach was removed, 5 cm of the small intestine was excised from the removed site, frozen in liquid nitrogen, and stored at −80 ° C.
Add 40 times the amount of extraction buffer (1 × PBS, 50 mM Tris-HCl (pH 6.8), Protease inhibitor cocktail (Roche cOmplete)) to the small intestine tissue, crush it in ice using a homogenizer, and remove the tissue lysate. Obtained. The tissue lysate was centrifuged at 10,000 × g for 10 minutes, and the supernatant was collected as a tissue extract and stored at −80 ° C.
The amount of IgA antibody in the tissue extract was measured using an ELISA kit (Bethyl Laboratories). At the same time, the protein concentration in the tissue extract was measured using a BCA kit (Thermo Scientific). The value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG.
１－２．試験結果
図１より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもタンパク質量当たりのIgA産生量が高いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の摂取によって、IgA抗体の産生が促進されることが示された。 1-2. Test results From FIG. 1, the group (SBT2171 group) ingesting the Lactobacillus helveticus SBT2171 strain (SBT2171 group) has higher IgA production per protein than the group (control group) ingesting the diet not added. I understood that. That is, it was shown that the production of IgA antibody is promoted by ingestion of Lactobacillus helveticus.
図１より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもタンパク質量当たりのIgA産生量が高いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の摂取によって、IgA抗体の産生が促進されることが示された。 1-2. Test results From FIG. 1, the group (SBT2171 group) ingesting the Lactobacillus helveticus SBT2171 strain (SBT2171 group) has higher IgA production per protein than the group (control group) ingesting the diet not added. I understood that. That is, it was shown that the production of IgA antibody is promoted by ingestion of Lactobacillus helveticus.
［試験例２］
人為的に自己抗体の産生を誘導して関節炎を発症させるコラーゲン誘発関節炎モデルマウスへの生菌ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株菌体の経口投与試験（IgG抗体量評価）
２－１．試験方法
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株をMRS培地で16時間培養した後、遠心分離により菌体を得た。菌体は滅菌生理食塩水で2回、超純水で1回洗浄した後、凍結乾燥した。凍結乾燥後の菌体は100 mg/mLとなるように0.35M NaHCO3/PBS に懸濁した後、使用するまで-20℃で保存した。
24匹のDBA/1Jマウス（8週齢、雄）を、コントロール（contorol）群とSBT2171群の2群に分けた（n=12）。ウシII型コラーゲン（CII）（2mg/ml in 0.05M acetic acid, Chondrex）とComplete Freund’s Adjuvant（CFA, Chondrex）を等量で合わせて免疫用のコラーゲンとして1mg/mlのエマルジョンを作製した。これを全てのマウスの尾根部に50 μLずつ2か所（合計100 μg/マウス1匹）に皮内投与した。1回目のコラーゲン投与日から21日後に2回目のコラーゲン投与を1回目と同様に行った。2回目のコラーゲン投与日から3週間飼育して解剖した。
SBT2171群には、1回目のコラーゲン投与日から解剖の前日まで毎日、100 mg/mLとなるように0.35M NaHCO3/PBSに懸濁したSBT217の菌体を、300 μL/マウス1匹（菌体量として30 mg/マウス1匹）となるように経口投与した。コントロール群には、0.35M NaHCO3/PBSを300 μL/マウス1匹となるように経口投与した。NaHCO3は、胃酸を中和する目的で添加した。
解剖時に採取した血液を遠心して血清を分離し、-80℃で保存した。血清中のCII特異的IgG抗体量を、ELISAキット（Chondrex）を用いて測定した。IgG抗体量を図２に示した。 [Test Example 2]
Oral administration test of Lactobacillus helveticus SBT2171 strain to collagen-induced arthritis model mice that artificially induce autoantibody production to develop arthritis (evaluation of IgG antibody amount)
2-1. Test Method Lactobacillus helveticus SBT2171 strain was cultured in MRS medium for 16 hours, and then the cells were obtained by centrifugation. The cells were washed twice with sterile physiological saline and once with ultrapure water, and then lyophilized. The cells after lyophilization were suspended in 0.35M NaHCO 3 / PBS so as to be 100 mg / mL, and stored at −20 ° C. until use.
Twenty-four DBA / 1J mice (8 weeks old, male) were divided into two groups, a control group and an SBT2171 group (n = 12). Bovine type II collagen (CII) (2 mg / ml in 0.05M acetic acid, Chondrex) and Complete Freund's Adjuvant (CFA, Chondrex) were combined in equal amounts to produce a 1 mg / ml emulsion as collagen for immunization. This was intradermally administered at two locations (total 100 μg / mouse) of 50 μL at the ridge of all mice. The second collagen administration was performed 21 days after the first collagen administration day in the same manner as the first administration. From the second collagen administration day, they were raised for 3 weeks and dissected.
In the SBT2171 group, SBT217 cells suspended in 0.35M NaHCO 3 / PBS at a concentration of 100 mg / mL were administered daily from the first collagen administration day to the day before dissection. It was orally administered so that the body weight was 30 mg / mouse. To the control group, 0.35M NaHCO 3 / PBS was orally administered so as to be 300 μL / mouse. NaHCO 3 was added for the purpose of neutralizing gastric acid.
Blood collected at the time of dissection was centrifuged to separate serum, and stored at -80 ° C. The amount of CII-specific IgG antibody in the serum was measured using an ELISA kit (Chondrex). The amount of IgG antibody is shown in FIG.
人為的に自己抗体の産生を誘導して関節炎を発症させるコラーゲン誘発関節炎モデルマウスへの生菌ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株菌体の経口投与試験（IgG抗体量評価）
２－１．試験方法
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株をMRS培地で16時間培養した後、遠心分離により菌体を得た。菌体は滅菌生理食塩水で2回、超純水で1回洗浄した後、凍結乾燥した。凍結乾燥後の菌体は100 mg/mLとなるように0.35M NaHCO3/PBS に懸濁した後、使用するまで-20℃で保存した。
24匹のDBA/1Jマウス（8週齢、雄）を、コントロール（contorol）群とSBT2171群の2群に分けた（n=12）。ウシII型コラーゲン（CII）（2mg/ml in 0.05M acetic acid, Chondrex）とComplete Freund’s Adjuvant（CFA, Chondrex）を等量で合わせて免疫用のコラーゲンとして1mg/mlのエマルジョンを作製した。これを全てのマウスの尾根部に50 μLずつ2か所（合計100 μg/マウス1匹）に皮内投与した。1回目のコラーゲン投与日から21日後に2回目のコラーゲン投与を1回目と同様に行った。2回目のコラーゲン投与日から3週間飼育して解剖した。
SBT2171群には、1回目のコラーゲン投与日から解剖の前日まで毎日、100 mg/mLとなるように0.35M NaHCO3/PBSに懸濁したSBT217の菌体を、300 μL/マウス1匹（菌体量として30 mg/マウス1匹）となるように経口投与した。コントロール群には、0.35M NaHCO3/PBSを300 μL/マウス1匹となるように経口投与した。NaHCO3は、胃酸を中和する目的で添加した。
解剖時に採取した血液を遠心して血清を分離し、-80℃で保存した。血清中のCII特異的IgG抗体量を、ELISAキット（Chondrex）を用いて測定した。IgG抗体量を図２に示した。 [Test Example 2]
Oral administration test of Lactobacillus helveticus SBT2171 strain to collagen-induced arthritis model mice that artificially induce autoantibody production to develop arthritis (evaluation of IgG antibody amount)
2-1. Test Method Lactobacillus helveticus SBT2171 strain was cultured in MRS medium for 16 hours, and then the cells were obtained by centrifugation. The cells were washed twice with sterile physiological saline and once with ultrapure water, and then lyophilized. The cells after lyophilization were suspended in 0.35M NaHCO 3 / PBS so as to be 100 mg / mL, and stored at −20 ° C. until use.
Twenty-four DBA / 1J mice (8 weeks old, male) were divided into two groups, a control group and an SBT2171 group (n = 12). Bovine type II collagen (CII) (2 mg / ml in 0.05M acetic acid, Chondrex) and Complete Freund's Adjuvant (CFA, Chondrex) were combined in equal amounts to produce a 1 mg / ml emulsion as collagen for immunization. This was intradermally administered at two locations (total 100 μg / mouse) of 50 μL at the ridge of all mice. The second collagen administration was performed 21 days after the first collagen administration day in the same manner as the first administration. From the second collagen administration day, they were raised for 3 weeks and dissected.
In the SBT2171 group, SBT217 cells suspended in 0.35M NaHCO 3 / PBS at a concentration of 100 mg / mL were administered daily from the first collagen administration day to the day before dissection. It was orally administered so that the body weight was 30 mg / mouse. To the control group, 0.35M NaHCO 3 / PBS was orally administered so as to be 300 μL / mouse. NaHCO 3 was added for the purpose of neutralizing gastric acid.
Blood collected at the time of dissection was centrifuged to separate serum, and stored at -80 ° C. The amount of CII-specific IgG antibody in the serum was measured using an ELISA kit (Chondrex). The amount of IgG antibody is shown in FIG.
２－２．試験結果
図２より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を摂取した群の方が、添加しない食餌を摂取した群よりもCII特異的IgG抗体産生量が低いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の摂取によって、自己抗体であるIgG抗体の産生が抑制されることが示された。 2-2. Test results From FIG. 2, it was found that the amount of CII-specific IgG antibody production was lower in the group ingesting the Lactobacillus helveticus SBT2171 strain than in the group ingested the diet not added. That is, it was shown that the production of IgG antibody which is an autoantibody is suppressed by ingestion of Lactobacillus helveticus.
図２より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を摂取した群の方が、添加しない食餌を摂取した群よりもCII特異的IgG抗体産生量が低いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の摂取によって、自己抗体であるIgG抗体の産生が抑制されることが示された。 2-2. Test results From FIG. 2, it was found that the amount of CII-specific IgG antibody production was lower in the group ingesting the Lactobacillus helveticus SBT2171 strain than in the group ingested the diet not added. That is, it was shown that the production of IgG antibody which is an autoantibody is suppressed by ingestion of Lactobacillus helveticus.
上記の試験例１、２により、ラクトバチルス・ヘルベティカス（Lactobacillus helvetivcus）を摂取することにより、IgA抗体産生の促進と自己抗体であるIgG抗体産生の抑制が可能となる事がわかった。
From the above Test Examples 1 and 2, it was found that by ingesting Lactobacillus helvetivcus, it is possible to promote the production of IgA antibody and suppress the production of IgG antibody which is an autoantibody.
[実施例１]ラクトバチルス・ヘルベティカスの培養物の製造
原料乳を加熱殺菌（75℃、15秒間）した後、30℃まで冷却し、0.01％塩化カルシウムを添加した。さらに、市販乳酸菌スターター（LDスターター、クリスチャン・ハンセン社）0.7％及びラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株（FERM BP-5445）1％を添加し、さらにレンネット0.003％を添加して、乳を凝固させた。このようにして得られた凝乳をカッティングし、ｐＨが6.2～6.1となるまで撹拌してホエーを排出して、カード粒を得た。そして、このカード粒を型詰めして圧搾し、さらに加塩して、10℃で熟成させ、ゴーダチーズタイプの硬質ナチュラルチーズを調製した。この硬質ナチュラルチーズ（6ヶ月熟成）をミンチ器（GM-DX、日本キャリア社製）を用いてミンチし、凍結乾燥を行った。その後、コーヒーミルにより微細化し、ラクトバチルス・ヘルベティカスの培養物であるチーズ粉を得た。該チーズ粉はそのまま本発明の抗体産生調節剤として使用できる。 [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. Furthermore, 0.7% of commercially available lactic acid bacteria starter (LD starter, Christian Hansen) and 1% of Lactobacillus helveticus SBT2171 strain (FERM BP-5445) were added, and 0.003% of rennet was further added. Solidified. The curd thus obtained was cut, stirred until the pH was 6.2 to 6.1, and whey was discharged to obtain curd grains. Then, the curd grains were filled and pressed, further salted, and aged at 10 ° C. to prepare gouda cheese type hard natural cheese. This hard natural cheese (aged for 6 months) was minced using a mincer (GM-DX, manufactured by Nippon Carrier Co., Ltd.) and freeze-dried. Then, it refined | miniaturized with the coffee mill and the cheese powder which is a culture of Lactobacillus helveticus was obtained. The cheese powder can be used as it is as an antibody production regulator of the present invention.
原料乳を加熱殺菌（75℃、15秒間）した後、30℃まで冷却し、0.01％塩化カルシウムを添加した。さらに、市販乳酸菌スターター（LDスターター、クリスチャン・ハンセン社）0.7％及びラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株（FERM BP-5445）1％を添加し、さらにレンネット0.003％を添加して、乳を凝固させた。このようにして得られた凝乳をカッティングし、ｐＨが6.2～6.1となるまで撹拌してホエーを排出して、カード粒を得た。そして、このカード粒を型詰めして圧搾し、さらに加塩して、10℃で熟成させ、ゴーダチーズタイプの硬質ナチュラルチーズを調製した。この硬質ナチュラルチーズ（6ヶ月熟成）をミンチ器（GM-DX、日本キャリア社製）を用いてミンチし、凍結乾燥を行った。その後、コーヒーミルにより微細化し、ラクトバチルス・ヘルベティカスの培養物であるチーズ粉を得た。該チーズ粉はそのまま本発明の抗体産生調節剤として使用できる。 [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. Furthermore, 0.7% of commercially available lactic acid bacteria starter (LD starter, Christian Hansen) and 1% of Lactobacillus helveticus SBT2171 strain (FERM BP-5445) were added, and 0.003% of rennet was further added. Solidified. The curd thus obtained was cut, stirred until the pH was 6.2 to 6.1, and whey was discharged to obtain curd grains. Then, the curd grains were filled and pressed, further salted, and aged at 10 ° C. to prepare gouda cheese type hard natural cheese. This hard natural cheese (aged for 6 months) was minced using a mincer (GM-DX, manufactured by Nippon Carrier Co., Ltd.) and freeze-dried. Then, it refined | miniaturized with the coffee mill and the cheese powder which is a culture of Lactobacillus helveticus was obtained. The cheese powder can be used as it is as an antibody production regulator of the present invention.
［試験例３］
マウスへのラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株培養物の経口投与試験（IgA抗体量）
１．試験方法
実施例１で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171の培養物を用いて経口投与試験を行った。試験方法は試験例１に準じて行った。解剖後のIgA抗体量とタンパク質濃度を測定し、IgA抗体量をタンパク質濃度で割った値を図３に示した。
２．試験結果
図３より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の培養物を添加した食餌を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもタンパク質量当たりのIgA産生量が高いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の培養物の摂取によって、IgA抗体の産生が促進されることが示された。 [Test Example 3]
Oral administration test of Lactobacillus helveticus SBT2171 culture to mice (IgA antibody level)
1. 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 IgA antibody amount and protein concentration after dissection were measured, and the value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG.
2. Test results From FIG. 3, the amount of protein in the group (SBT2171 group) ingesting the diet supplemented with the culture of Lactobacillus helveticus SBT2171 strain was greater than that in the group ingesting the diet not added (control group) It was found that the amount of IgA production per unit was high. That is, it was shown that the production of IgA antibody is promoted by ingestion of a culture of Lactobacillus helveticus.
マウスへのラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株培養物の経口投与試験（IgA抗体量）
１．試験方法
実施例１で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171の培養物を用いて経口投与試験を行った。試験方法は試験例１に準じて行った。解剖後のIgA抗体量とタンパク質濃度を測定し、IgA抗体量をタンパク質濃度で割った値を図３に示した。
２．試験結果
図３より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の培養物を添加した食餌を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもタンパク質量当たりのIgA産生量が高いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の培養物の摂取によって、IgA抗体の産生が促進されることが示された。 [Test Example 3]
Oral administration test of Lactobacillus helveticus SBT2171 culture to mice (IgA antibody level)
1. 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 IgA antibody amount and protein concentration after dissection were measured, and the value obtained by dividing the IgA antibody amount by the protein concentration is shown in FIG.
2. Test results From FIG. 3, the amount of protein in the group (SBT2171 group) ingesting the diet supplemented with the culture of Lactobacillus helveticus SBT2171 strain was greater than that in the group ingesting the diet not added (control group) It was found that the amount of IgA production per unit was high. That is, it was shown that the production of IgA antibody is promoted by ingestion of a culture of Lactobacillus helveticus.
［試験例４］
マウスへのラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株培養物の経口投与試験（IgG抗体量）
１．試験方法
実施例１で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171の培養物を用いて経口投与試験を行った。試験方法は試験例２に準じて行った。解剖後の血清中のCII特異的IgG抗体量を測定し、その結果を図４に示した。
２．試験結果
図４より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の培養物を添加した食餌を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもCII特異的IgG抗体産生量が低いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の培養物の摂取によって、自己抗体であるIgG抗体の産生を抑制することが示された。 [Test Example 4]
Oral administration test of Lactobacillus helveticus SBT2171 strain culture to mice (IgG antibody level)
1. 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 2. The amount of CII-specific IgG antibody in the serum after dissection was measured, and the results are shown in FIG.
2. Test results From FIG. 4, the group that ingested the diet supplemented with the culture of Lactobacillus helveticus SBT2171 (SBT2171 group) was more CII-specific than the group that ingested the diet not added (control group) It was found that the production amount of IgG antibody was low. That is, it was shown that the ingestion of a culture of Lactobacillus helveticus suppresses the production of IgG antibodies that are autoantibodies.
マウスへのラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株培養物の経口投与試験（IgG抗体量）
１．試験方法
実施例１で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171の培養物を用いて経口投与試験を行った。試験方法は試験例２に準じて行った。解剖後の血清中のCII特異的IgG抗体量を測定し、その結果を図４に示した。
２．試験結果
図４より、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の培養物を添加した食餌を摂取した群（SBT2171群）の方が、添加しない食餌を摂取した群（コントロール群）よりもCII特異的IgG抗体産生量が低いことが分かった。すなわち、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）の培養物の摂取によって、自己抗体であるIgG抗体の産生を抑制することが示された。 [Test Example 4]
Oral administration test of Lactobacillus helveticus SBT2171 strain culture to mice (IgG antibody level)
1. 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 2. The amount of CII-specific IgG antibody in the serum after dissection was measured, and the results are shown in FIG.
2. Test results From FIG. 4, the group that ingested the diet supplemented with the culture of Lactobacillus helveticus SBT2171 (SBT2171 group) was more CII-specific than the group that ingested the diet not added (control group) It was found that the production amount of IgG antibody was low. That is, it was shown that the ingestion of a culture of Lactobacillus helveticus suppresses the production of IgG antibodies that are autoantibodies.
[実施例２] 抗体産生調節剤（顆粒）の製造
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を食用可能な合成培地（0.5％酵母エキス、0.1％トリプチケースペプトン添加）に5重量％接種し、38℃で15時間培養後、遠心分離で菌体を回収した。回収した菌体を凍結乾燥し、前記菌体の凍結乾燥粉末を得た。この凍結乾燥粉末1 gを乳糖5 gと混合し、顆粒状に成形して本発明の抗体産生調節剤を得た。 [Example 2] Production of antibody production regulator (granule) Lactobacillus helveticus SBT2171 strain was inoculated into 5% by weight in an edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added). After 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 antibody production regulator of the present invention.
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株を食用可能な合成培地（0.5％酵母エキス、0.1％トリプチケースペプトン添加）に5重量％接種し、38℃で15時間培養後、遠心分離で菌体を回収した。回収した菌体を凍結乾燥し、前記菌体の凍結乾燥粉末を得た。この凍結乾燥粉末1 gを乳糖5 gと混合し、顆粒状に成形して本発明の抗体産生調節剤を得た。 [Example 2] Production of antibody production regulator (granule) Lactobacillus helveticus SBT2171 strain was inoculated into 5% by weight in an edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added). After 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 antibody production regulator of the present invention.
[実施例３] 抗体産生調節剤（顆粒）の製造
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）JCM-1120株を食用可能な合成培地（0.5％酵母エキス、0.1％トリプチケースペプトン添加）に5重量％接種し、38℃で15時間培養後、遠心分離で菌体を回収した。回収した菌体を凍結乾燥し、前記菌体の凍結乾燥粉末を得た。この凍結乾燥粉末1 gを乳糖5 gと混合し、顆粒状に成形して本発明の抗体産生調節剤を得た。 [Example 3] Manufacture of antibody production regulator (granule) 5% by weight of Lactobacillus helveticus JCM-1120 strain in edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added) After inoculation 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 antibody production regulator of the present invention.
ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）JCM-1120株を食用可能な合成培地（0.5％酵母エキス、0.1％トリプチケースペプトン添加）に5重量％接種し、38℃で15時間培養後、遠心分離で菌体を回収した。回収した菌体を凍結乾燥し、前記菌体の凍結乾燥粉末を得た。この凍結乾燥粉末1 gを乳糖5 gと混合し、顆粒状に成形して本発明の抗体産生調節剤を得た。 [Example 3] Manufacture of antibody production regulator (granule) 5% by weight of Lactobacillus helveticus JCM-1120 strain in edible synthetic medium (0.5% yeast extract, 0.1% trypticase peptone added) After inoculation 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 antibody production regulator of the present invention.
［実施例４］抗体産生調節剤（散剤）の製造
第13改正日本薬局方解説書製剤総則「散剤」の規定に準拠し、上記実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末10 gに乳糖（日局）40 g、バレイショデンプン（日局）600 gを加えて均一に混合し、本発明の抗体産生調節剤を製造した。 [Example 4] Manufacture of antibody production regulator (powder) The Lactobacillus helveticus SBT2171 obtained in Example 2 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder" Lactose (JP) 40 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 antibody production regulator of the present invention.
第13改正日本薬局方解説書製剤総則「散剤」の規定に準拠し、上記実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末10 gに乳糖（日局）40 g、バレイショデンプン（日局）600 gを加えて均一に混合し、本発明の抗体産生調節剤を製造した。 [Example 4] Manufacture of antibody production regulator (powder) The Lactobacillus helveticus SBT2171 obtained in Example 2 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder" Lactose (JP) 40 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 antibody production regulator of the present invention.
［実施例５］抗体産生調節剤（散剤）の製造
第13改正日本薬局方解説書製剤総則「散剤」の規定に準拠し、上記実施例３で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）JCM-1120株の凍結乾燥粉末10 gに乳糖（日局）400 g、バレイショデンプン（日局）600 gを加えて均一に混合し、本発明の抗体産生調節剤を製造した。 [Example 5] Manufacture of antibody production regulator (powder) The Lactobacillus helveticus JCM obtained in Example 3 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder" Lactose (JP) 400 g and potato starch (JP) 600 g were added to 10 g of freeze-dried powder of strain -1120 and mixed uniformly to produce the antibody production regulator of the present invention.
第13改正日本薬局方解説書製剤総則「散剤」の規定に準拠し、上記実施例３で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）JCM-1120株の凍結乾燥粉末10 gに乳糖（日局）400 g、バレイショデンプン（日局）600 gを加えて均一に混合し、本発明の抗体産生調節剤を製造した。 [Example 5] Manufacture of antibody production regulator (powder) The Lactobacillus helveticus JCM obtained in Example 3 above in accordance with the provisions of the 13th revised Japanese Pharmacopoeia Guideline General Formulation "Powder" Lactose (JP) 400 g and potato starch (JP) 600 g were added to 10 g of freeze-dried powder of strain -1120 and mixed uniformly to produce the antibody production regulator of the present invention.
［実施例６］スティック状栄養健康食品の製造
ビタミンＣ40 gまたはビタミンＣとクエン酸の等量混合物40 g、グラニュー糖100 g、コーンスターチと乳糖の等量混合物60 gに、上記実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末40 gを加えて混合した。混合物を袋に詰め、本発明のスティック状栄養健康食品を150袋製造した。 [Example 6] Manufacture of stick-like nutritional health foods Vitamin C 40 g or an equivalent mixture of vitamin C and citric acid 40 g, granulated sugar 100 g, corn starch and lactose equivalent mixture 60 g obtained in Example 2 above 40 g of the lyophilized powder of the obtained Lactobacillus helveticus SBT2171 strain 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.
ビタミンＣ40 gまたはビタミンＣとクエン酸の等量混合物40 g、グラニュー糖100 g、コーンスターチと乳糖の等量混合物60 gに、上記実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株の凍結乾燥粉末40 gを加えて混合した。混合物を袋に詰め、本発明のスティック状栄養健康食品を150袋製造した。 [Example 6] Manufacture of stick-like nutritional health foods Vitamin C 40 g or an equivalent mixture of vitamin C and citric acid 40 g, granulated sugar 100 g, corn starch and lactose equivalent mixture 60 g obtained in Example 2 above 40 g of the lyophilized powder of the obtained Lactobacillus helveticus SBT2171 strain 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.
［実施例７］飼料の製造
大豆粕12 kg、脱脂粉乳14 kg、大豆油4 kg、コーン油2 kg、パーム油23.2kg、トウモロコシ澱粉14 kg、小麦粉9 kg、ふすま2 kg、ビタミン混合物5 kg、セルロース2.8 kg、ミネラル混合物2 kgを配合し、120℃、4分間殺菌して、実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株10 kgを配合して、飼料を製造した。 [Example 7] Production offeed 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 mineral mixture were 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. .
大豆粕12 kg、脱脂粉乳14 kg、大豆油4 kg、コーン油2 kg、パーム油23.2kg、トウモロコシ澱粉14 kg、小麦粉9 kg、ふすま2 kg、ビタミン混合物5 kg、セルロース2.8 kg、ミネラル混合物2 kgを配合し、120℃、4分間殺菌して、実施例２で得られたラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171株10 kgを配合して、飼料を製造した。 [Example 7] Production of
本発明により、ウイルスや細菌などに対する感染予防効果とともに、過剰な自己免疫応答を予防、改善することができる。さらに、該乳酸菌を抗含有するチーズまたは乳酸菌飲料を食することで、抗体産生を調節することも可能となる
According to the present invention, an excessive autoimmune response can be prevented and improved together with an infection preventing effect against viruses and bacteria. Furthermore, it is also possible to regulate antibody production by eating cheese or lactic acid bacteria beverages that are anti-containing the lactic acid bacteria.
[寄託生物材料への言及]
（１）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 産業技術総合研究所 特許生物寄託センター
日本国茨城県つくば市東１丁目１番3号
ロ イの寄託機関に生物材料を寄託した日付
平成６年６月２２日（１９９４年６月２２日）（原寄託日）
平成８年３月６日（１９９６年３月６日）（原寄託によりブタペスト条約に基づく寄託へ
の移管日）
ハ イの寄託機関が寄託について付した受託番号
FERM BP-5445 [Reference to deposited biological materials]
(1) 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
（１）ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 産業技術総合研究所 特許生物寄託センター
日本国茨城県つくば市東１丁目１番3号
ロ イの寄託機関に生物材料を寄託した日付
平成６年６月２２日（１９９４年６月２２日）（原寄託日）
平成８年３月６日（１９９６年３月６日）（原寄託によりブタペスト条約に基づく寄託へ
の移管日）
ハ イの寄託機関が寄託について付した受託番号
FERM BP-5445 [Reference to deposited biological materials]
(1) 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
Claims (6)
- ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。 An antibody production regulator that promotes the production of an IgA antibody comprising a lactic acid bacterium belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of an IgG antibody that is an autoantibody.
- 前記ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌がラクトバチルス・ヘルベティカス（Lactobacillus helveticus）SBT2171（FERM BP-5445）であることを特徴とする、請求項１に記載の IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤。 The lactic acid bacterium belonging to Lactobacillus helveticus (Lactobacillus helveticus) is Lactobacillus helveticus SBT2171 (FERM BP-5445), which promotes production of IgA antibody according to claim 1, An antibody production regulator that suppresses the production of IgG antibodies that are autoantibodies.
- 請求項１または請求項２に記載の剤を添加したIgG抗体の産生を抑制する抗体産生調節用栄養組成物、飲食品又は飼料。 A nutritional composition for regulating antibody production, food or drink, or feed that suppresses the production of IgG antibodies to which the agent according to claim 1 or 2 is added.
- ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する抗体産生調節剤の使用方法。 An antibody that administers to a subject an antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies How to use production regulators.
- ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物を有効成分とするIgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤を対象に投与する感染症の予防と自己免疫疾患の予防のための前記抗体産生調節剤の使用方法。 Infection by administering to the subject an antibody production regulator that promotes the production of IgA antibodies containing lactic acid bacteria belonging to Lactobacillus helveticus or its culture as an active ingredient and suppresses the production of IgG antibodies that are autoantibodies Use of the antibody production regulator for prevention of infectious diseases and autoimmune diseases.
- IgA抗体の産生を促進し、かつ自己抗体であるIgG抗体の産生を抑制する抗体産生調節剤の製造における、ラクトバチルス・ヘルベティカス（Lactobacillus helveticus）に属する乳酸菌又はその培養物の使用。
Use of a lactic acid bacterium belonging to Lactobacillus helveticus or a culture thereof in the production of an antibody production regulator that promotes production of IgA antibody and suppresses production of IgG antibody that is an autoantibody.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2015-189886 | 2015-09-28 | ||
| JP2015189886A JP6739155B2 (en) | 2015-09-28 | 2015-09-28 | Antibody production regulator |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2017057307A1 true WO2017057307A1 (en) | 2017-04-06 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/JP2016/078337 WO2017057307A1 (en) | 2015-09-28 | 2016-09-27 | Antibody production controller |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP6739155B2 (en) |
| WO (1) | WO2017057307A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019210242A (en) * | 2018-06-04 | 2019-12-12 | 雪印メグミルク株式会社 | Srcap expression-inhibiting compositions containing lactic acid bacteria of the genus lactobacillus as an effective ingredient |
-
2015
- 2015-09-28 JP JP2015189886A patent/JP6739155B2/en active Active
-
2016
- 2016-09-27 WO PCT/JP2016/078337 patent/WO2017057307A1/en active Application Filing
Non-Patent Citations (4)
| Title |
|---|
| FUMIHIKO SAKAI ET AL.: "Lactobacillus gasseri SBT2055 induces TGF-beta expression in dendritic cells and activates TLR2 signal to produce IgA in the small intestine", PLOS ONE, vol. 9, no. 8, 2014, pages 1 - 11, XP055371441 * |
| JIA YU -CHEN ET AL.: "Effects of Lactobacillus helveticus on Immune Response and Cytokines in Intestinal Mucosa of Mice", FOOD SCIENCE, vol. 31, no. 23, 2010, pages 335 - 9 * |
| JUNG-EUN KIMA ET AL.: "Lactobacillus helveticus suppresses experimental rheumatoid arthritis by reducing inflammatory T cell responses", JOURNAL OF FUNCTIONAL FOODS, vol. 13, February 2015 (2015-02-01), pages 350 - 362, XP055371439 * |
| TOMOHIRO HOSOYA ET AL.: "Lactobacillus helveticus SBT2171 inhibits lymphocyte proliferation by regulation of the JNK signaling pathway", PLOS ONE, vol. 9, no. 9, 2014, pages 1 - 9, XP055371434 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019210242A (en) * | 2018-06-04 | 2019-12-12 | 雪印メグミルク株式会社 | Srcap expression-inhibiting compositions containing lactic acid bacteria of the genus lactobacillus as an effective ingredient |
| JP7224114B2 (en) | 2018-06-04 | 2023-02-17 | 雪印メグミルク株式会社 | Composition for suppressing SRCAP expression containing lactic acid bacteria of the genus Lactobacillus as an active ingredient |
Also Published As
| Publication number | Publication date |
|---|---|
| JP6739155B2 (en) | 2020-08-12 |
| JP2017066047A (en) | 2017-04-06 |
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