KR100532362B1 - Bifidobacterium infantis which prevent and/or cure allergic diseases - Google Patents

Abstract

The present invention relates to lactic acid bacteria that can prevent or inhibit allergic diseases, Lactobacillus coprophilus Strain PL9001 (KCCM-10245), Lactobacillus fermentum Strain PL9005 (Lactobacillus fermentum Strain PL9005, KCCM-10250), Lactobacillus plantarum PL9011 (KCCM-10358) and Bifidobacterium infantis PL9506 (Bifidobacterium infantis PL9506, KCCM-10406). The strains of the present invention can be used as a prophylactic and therapeutic agent for various allergic diseases, and can be used directly or as additives in the preparation of allergic disease preventive and therapeutic agents.

Description

Bifidobacterium infantis which prevent and / or cure allergic diseases}

The present invention provides lactic acid bacteria that can prevent allergic diseases.

The present invention also provides lactic acid bacteria that can treat allergic diseases.

In another aspect, the present invention provides a food additive capable of suppressing allergies, including live bacteria, dead bacteria, crushed medicine, culture solution, culture extract of the present lactic acid bacteria.

In another aspect, the present invention provides a food and beverage having an anti-allergic function using lactic acid bacteria having an allergic inhibitory effect as a seed.

The present invention also provides a cosmetic containing lactic acid bacteria having an allergic inhibitory effect.

The present invention also provides a skin ointment containing lactic acid bacteria having an allergic inhibitory effect.

The present invention also provides a skin adhesive containing lactic acid bacteria having an allergic inhibitory effect.

The present invention also provides a dose containing lactic acid bacteria having an allergic inhibitory effect.

The rise in allergic diseases over the last few decades is explained by changes in the environment, along with decreased microbial exposure and changes in food intake.

Preventing an increase in the incidence of allergic diseases has become a major problem for the health of Western societies. The increase in allergic diseases is attributed to reduced exposure to microorganisms at the beginning of life due to improvements in the living environment, according to The hygiene hypothesis.

The first systematic study of allergy phenomena was the French physiologist Riche, who described anaphylaxis in 1902 and the next year, the French physiologist Arthus reported about the arthrosis (local anaphylaxis). After several injections of anemone toxin into his dog, Riche re-injected the same toxin and found that he died due to the distinctive sudden symptoms at a much lower dose than the initial dose. It means no defense. At first, it was regarded as a unique phenomenon of toxins, but afterwards, the same phenomenon was observed in other proteins.

The origin of the allergy is a compound word of Greek meaning 'dichroic effect (異 色 作用)'. In other words, if you have some kind of infectious disease, or pretreatment with a bacterial product or a heterogeneous substance, pay attention to the constant change in the body's reaction ability. It was considered that the phenomenon occurs to produce antibodies in vivo. Richet is also known to have applied this idea to the clinical practice of tuberculosis and applied tuberculin-specific skin reactions to the diagnosis of tuberculosis.

On the other hand, Artus was injected with a heterologous protein in rabbits and reinjected a small amount of the same substance subcutaneously after a period of time, and found that an inflammatory reaction accompanied by edema or necrosis appeared locally. In addition to these experimental facts, Riche shows a faster and stronger skin reaction in the second head of the person than in the first time, and in the serum disease the first symptoms appear after about a week, after the second time immediately after injection. The concept of allergy was introduced on the basis of extensive clinical observations, such as symptoms.

In other words, in addition to anaphylaxis, serum diseases, and so-called specific constitutions, all of them are considered to be biological reactions by antigen-antibody reactions, including immune phenomena once the concept is clarified. The weakened reaction capacity is called sensitivity, insensitivity or immunity.

However, biological reactions due to antigen-antibody reactions are a complex phenomenon, and because of the fact that their mechanisms are unknown, many definitions and terminology, such as allergies or atopy, have been published. woke up.

Currently, allergens define pathological processes among the effects of antigen-antibody responses on living organisms. In other words, clinically, hyperactivity (hypersensitivity) is strongly indicated on the surface, so allergy is almost synonymous with hypersensitivity.

Sterile animal experiments have shown that the gut microbiota plays an important role in triggering the immune system to suppress various reactions, including allergies. In clinical trials, we also found a high rate of allergy in children with a small number of lactic acid-producing bacteria, such as intestinal lactic acid bacteria and nasal bacteria. It was also confirmed that the gut bacteria regulate immune atopy. On the other hand, there is some evidence that probiotic added foods suppress allergy and atopic development. Recent studies have shown that several lactic acid bacteria stimulate the production of type I and II interferons and pro-interferon monokines (IL-12 and IL-18). Thus, lactic acid bacteria of immunomodulatory activity can be used as food additives (adjuvant) to change the intestinal flora, thereby providing sufficient pro-T helper cell 1 (Th1) to correct Th2-type to restore immune phenomena and promote allergic reactions. Can provide STAT-activating signals (Cross ML, Gill HS. 2001. Can immunoregulatory lactic acid bacteria be used as dietary supplements to limit allergies? Int. Arch. Allergy Immunol. 2001 Jun; 125 (2): 112-9 Review.)

Clinical trials have shown that ingesting fermented foods, such as yogurt, can alleviate the symptoms of atopy and reduce the development of allergies by immunomodulation. Intake of fermented milk containing lactic acid bacteria improved production of Type I and Type II interferons as a whole. In animal experiments, lactic acid bacteria promoted interferon expression and reduced the production of IL-4 and IL-5 by allergens. Recent results suggest that lactic acid bacteria strongly induce pro-interferon monokines (IL-12 and IL-18), which are stimulated by the interaction of receptors and cell walls on the surface of leukocytes (mononuclear phagocytes) with NF-kappa B and STAT signaling It is caused by pathways. However, the degree and quality of immune regulation induced by lactic acid bacteria depends on strain (Cross ML, Stevenson LM, Gill HS. 2001. Anti-allergy properties of fermented foods: an important immunoregulatory mechanism of lactic acid bacteria? Int Immunopharmacol 1 ( 5): 891-901).

Allergic reactions are thought to result from the lack of exposure to microorganisms that maintain mutual balance in childhood. The composition of the first intestinal flora is considered to be an important determinant in the development of immune response forms and normal intestinal defense. Experimental, clinical and pathological studies over the past decades have shown that non-pathogenic gut bacteria are essential factors for the body's immune system to mature in a nontopic manner.

In allergic diseases, the regulatory effects of probiotics have been proven to improve the therapeutic effects or prevent the occurrence of Atopic exima in children with probiotic added foods. This means that gram-negative lipopolysaccharide and CpG from bacterial DNA activate immunoregulatory genes through toll-like receptors in the intestinal epidermal cells. Probiotics also act to degrade and alter the polymer to reduce antigens in the gut. (Kalliomaki M, Isolauri E. 2002. Pandemic of atopic diseases--a lack of microbial exposure in early infancy? Curr Drug Targets Infect Disord 2 (3): 193-9). Probiotic foods also allow gut-associated lymphoid tissue (GALT), which is often deficient in infants, to mature. Probiotic nutrition normalizes intestinal permeability, balances intestinal bacterial flora, improves intestinal immune defense mechanisms (IgA), alleviates intestinal inflammatory responses, and inhibits cytokines, a hallmark of local and general allergic inflammation (Miraglia del Giudice M Jr, De Luca MG, Capristo C. 2002. Probiotics and atopic dermatitis.A new strategy in atopic dermatitis. Dig Liver Dis 34 Suppl 2: S68-71). Some lactic acid probiotics have inhibitory effects on allergic inflammation, which is thought to be involved in proteolytic and immunomodulatory activities (von der WT, Ibnou-Zekri N, Pfeifer A. 2002. Novel probiotics for the management of allergic inflammation.Dig Liver Dis. 34 Suppl 2: S25-8). Therefore, scientifically formulated functional foods containing probiotics and other functional elements are expected to help prevent and control allergic diseases (Laiho K, Ouwehand A, Salminen S, Isolauri E. 2002. Inventing probiotic functional foods for patients with allergic disease.Ann. Allergy Asthma Immunol. 89 (6 Suppl 1): 75-82).

Atopic disease is said to be caused by the lack of exposure to microorganisms that equilibrate early in life. In other words, the initial composition of the intestinal flora is considered to be an important determinant of the development of immune response and normal intestinal inhibitory action. The probiotic regulatory role in human allergic diseases was first emphasized in the laboratory by showing inhibitory effects on leukocyte proliferation and IL-4, and probiotic supplementation was effective in infants' atopic eczema. (Laiho K, Hoppu U, Ouwehand AC, Salminen S, Isolauri E. 2002. Probiotics: on-going research on atopic individuals.Br J Nutr 2002 88 Suppl 1: S19-27)

In clinical trials, probiotic administration in pregnant and lactating women increased the immune defense function of breast milk. When not administered, the amount of anti-inflammatory transforming growth factor beta2 (TGF-beta2) in breast milk was 1,340 pg / mL [95% CI, 978-1702, compared with 2885 pg / mL [95% CI, 1624-4146]. In probiotic mothers, the development of Atopic eczema during the first two years of the newborn was significantly reduced compared to the baby of the mother who received a placebo (15% and 47%, respectively; relative risk, 0.32 [95% CI, 0.12-0.85]; P = .0098). Mother's atopy is the risk factor for atopic eczema in newborns. The most virtue of probiotic prescription was a high concentration of IgE in the umbilical cord. Administration of probiotics during pregnancy and lactation is a safe, effective and protective effect on atopic eczema in the first two years of life (Rautava S, Kalliomaki M, Isolauri E. 2002. Probiotics during pregnancy and breast-feeding might confer immunomodulatory protection against atopic disease in the infant.J Allergy Clin Immunol 109 (1): 119-21). When relatives or husbands received Lactobacillus GG for mothers and newborns with atopic eczema, allergic rhinitis, and asthma for six months, the incidence of atopic eczema was reduced by half compared to the non-administered group (Kalliomaki M, Salminen S). , Arvilommi H, Kero P, Koskinen P, Isolauri E. 2001. Probiotics in primary prevention of atopic disease: a randomised placebo-controlled trial.Lancet 357 (9262): 1076-1079). Lactobacillus GG (ATCC 53103) regulated antigen adsorption by increasing local antigen-specific immune responses (particularly IgA). Fecal alpha 1-antitrypsin, tumor necrosis factor-alpha, and eosinophil cationic protein in feces are administered to mothers who add or feed Lactobacillus GG to lactated skim milk in newborns allergic to atopic eczema and milk. Was measured. In newborns fed hydrolyzed skim milk supplemented with Lactobacillus GG, atopic dermatitis was significantly improved in one month of experiments. The amount of alpha 1-antitrypsin was decreased and fecal tumor necrosis factor-alpha was significantly decreased. However, the amount of fecal eosinophil cationic protein did not change. These results have shown that probiotics can be used to treat intestinal defense mechanisms and reduce intestinal inflammation in patients with atopic dermatitis and food allergies (Majamaa H, Isolauri E. 1997. Probiotics: a novel approach in the management of food allergy.J Allergy Clin Immunol 99 (2): 179-85).

Probiotic research is currently underway to identify specific strains with antiallergic functions (Isolauri E, Rautava S, Kalliomaki M, Kirjavainen P, Salminen S. 2002. Role of probiotics in food hypersensitivity.Cur Opin Allergy Clin Immunol 2 (3): 263-71).

Therefore, an object of the present invention is to provide a strain capable of preventing allergies.

The strains are Lactobacillus coprophilus Strain PL9001, KCCM-10245, Lactobacillus fermentum Strain PL9005, KCCM-10250, Lactobacillus plantarum Strain PL9011, KCCM-10 And Bifidobacterium infantis Strain PL9506 (KCCM-10406), these lactic acid bacteria have been found to have Helicobacter pylori growth and adhesion inhibition, and the patented strains, Lactobacillus copropylus PL9001 KCCM-10245, Lactobacillus Fermentum PL9005 is KCCM-10250, Lactobacillus Plantarum PL9011 is KCCM-10358, Bifidobacterium Infantis PL9506 is KCCM-10406, and these strains are Helicobacter pylori and It has been found to have intestinal bacterial growth and intestinal epithelial cell adhesion inhibition.

It is another object of the present invention to provide a lactic acid bacterium capable of treating allergic diseases.

It is another object of the present invention to provide a lactic acid bacterium capable of preventing allergies caused by various allergens in the air.

It is another object of the present invention to provide a lactic acid bacterium capable of preventing allergies caused by various food allergens.

It is another object of the present invention to provide a lactic acid bacterium that can prevent or treat atopic dermatitis.

In another aspect, the present invention is to provide a lactic acid bacteria that can prevent or treat allergic rhinitis, asthma, atopic eczema.

The lactic acid bacteria can be formulated into various compositions. Preferably, the composition is also preferably in the form of pharmaceutical compositions such as capsules, tablets, and powders, and in forms easy to add various foods. Such formulations may be prepared using known pharmaceutically acceptable carriers, excipients, solvents or auxiliaries by known methods. Such methods and components are well known and are described in detail in standard texts and manuals, such as, for example, Remington. 1995. The Science and Practice of Pharmacy. Mack Publising Co. Easton, PA 18042, USA. It is.

In addition, formal bacteria can be prepared in food by methods generally known in the art.

It can also be used as a seed for fermented foods, including fermented dairy products, by methods well known in the art to reduce food allergens.

Hereinafter, the present invention will be described in detail.

In order to achieve the above object, the present invention provides an allergy prevention agent and a therapeutic agent.

In addition, the present invention is Lactobacillus coprophilus Strain PL9001, KCCM-10245, Lactobacillus fermentum Strain PL9005 (KCCM-10250), Lactobacillus plantarum PL9011 (Lactobacillus) for allergy prevention and treatment plantarum Strain PL9011, KCCM-10358) and Bifidobacterium infantis PL9506 (Bifidobacterium infantis Strain PL9506, KCCM-10406).

In addition, the present invention is Lactobacillus copropylus PL9001 (KCCM-10245), Lactobacillus Fermentum PL9005 (KCCM-10250), Lactobacillus plantarum PL9011 (KCCM-10358), Bifidobacterium infantis for allergy prevention and treatment Providing allergy prevention and therapeutic oral medications in which one or more of PL9506 (KCCM-10406) is taken from 1 × 10 ⁷ CFU to 1 × 10 ¹² CFU 1-3 times a day.

In another aspect, the present invention is Lactobacillus coprophilus Strain PL9001, KCCM-10245, Lactobacillus fermentum Strain PL9005, KCCM-10250), Lactobacillus plantarum Strain PL9011 (Lactobacillus plantarum Strain PL9011) 10358) and Bifidobacterium infantis Strain PL9506 (KCCM-10406), including external skin preparations including functional cosmetics for allergy prevention and treatment purposes.

In another aspect, the present invention is Lactobacillus coprophilus Strain PL9001, KCCM-10245, Lactobacillus fermentum Strain PL9005, KCCM-10250), Lactobacillus plantarum Strain PL9011 (Lactobacillus plantarum Strain PL9011) 10358) and Bifidobacterium infantis Strain PL9506, KCCM-10406).

In another aspect, the present invention is Lactobacillus coprophilus Strain PL9001, KCCM-10245, Lactobacillus fermentum Strain PL9005, KCCM-10250), Lactobacillus plantarum Strain PL9011 (Lactobacillus plantarum Strain PL9011) 10358) and Bifidobacterium infantis Strain PL9506 (KCCM-10406), which provide fermentation products for allergy prevention and treatment purposes.

Hereinafter, preferred embodiments of the present invention are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the present invention is not limited to the following examples.

Experimental Example 1 Passive Cutaneous Anaphylaxis (PCA) test

As an animal model, Ovalbumin was administered in mice to induce systemic hypersensitivity reactions, and serum was isolated at 21 days after inoculation into the skin of rats, and 30 minutes later, ovalbumin was readministered through the microvenous vein. Evans blue solution was used as an indicator of skin hypersensitivity. Using the experimental animal model constructed using this method, the effect of each test substance on the skin hypersensitivity reaction was evaluated.

Materials and methods

Test substance

Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9506 (KCCM-10406) were lyophilized and stored under refrigeration under sealed conditions. Diluted in sterile PBS before administration of the test material was incubated for 1 hour in a 37 ℃ Shaking incubator.

Laboratory animals

50 6-week-old Wister Rats and 3 6-week-old BALB / c mice were obtained from Charles River (Japan). Rats were set high and low concentration groups for each test substance, including negative and positive controls (groups administered with PBS instead of test substance). All experiments in an animal room controlled with temperature 22 ± 3 ℃, relative humidity 50 ± 10%, number of ventilation 10-12 times / hr, lighting time 12 hours (08: 00 ~ 20: 00), and illuminance 150-200 lux The animals were bred for a while, and the water and feed were freely supplied.

Induction of systemic anaphylaxis in mice

Three mice were mixed with 50 µl Ovalbumin (Sigma: grade V) and 0.2 ml of 2 mg of Al (OH) ₃ as inducers of the hypersensitivity reaction, and were administered intraperitoneally once every 0 and 14 days of the experiment.

IgE measurement in sensitized mouse serum.

On day 0, day 14, and day 20 of the experiment, blood was collected from the intraorbital venous plexus, and the serum was obtained by centrifugation at 3000 rpm for 20 minutes after refrigeration for 2 hours. The serum IgE sandwich ELISA kit (Shibayagi, Japan) was used to measure the amount of IgE expressed in the serum mixed with each serum and serum collected from each individual on day 20.

Administration of Test Substances in Rats

The experimental group contained Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), Bifidobacterium infantis PL9506 (KCCM-10406) for 21 days and 1 × 10 ⁹ CFU / ml. Quantitatively, 1 × 10 ⁷ CFU / ml was administered orally once a day between 10 am and 12 am using zondae.

Negative and positive controls were administered with sterile phosphate buffered saline (PBS) instead of the test substance.

Induction of skin sensitization in rats

The combined serum from three mice was diluted 10-fold with PBS, and intradermal injection was performed at 100 μl each to the back of the rats from which hair was removed. PBS was administered to the negative control group. After 30 minutes, ovalbumin 100 ㎍ / 0.5mL and 0.5% Evans blue solution (Sigma) were mixed and administered intravenously.

Measurement of ovalbumin-specific IgE in Rat Skin

After induction of skin sensitization in rats, euthanasia was performed using ether after 2 hours, and 0.15 g of skin tissue of the administered portion was quantified. To extract Evans blue, skin tissue collected in 400 μl of Formamide (Sigma) was deposited for 48 hours, and the absorbance (Optical Density, OD) value was measured at 650 nm using an ELISA reader.

Statistical treatment

Duncan's multiple test of SAS was used to test group significance.

result

Body weight and feed intake

Body weight and feed intake of rats increased from the beginning to the end of the group without any significant difference.

Serum IgE in Mice Induced Systemic Sensitization

On the 20th day, the amount of IgE in the serum was increased 8 to 10 times compared to the 0 day of Ovalbumin administration. To induce hypersensitivity in rats, the serum IgE of serum from three mice on day 20 of the test was 138.05 ng / ml (Table 1).

Table 1-1 Changes in IgE Concentrations in Anaphylaxy-induced Mouse Serum

Determination of Skin Sensitization in Rats

To induce hypersensitivity, the antigen-inoculated site was blue due to the deposition of Evans blue when visually observed. In this experiment, Evans blue is a measure of the combined amount of ovalbumin and specific IgE. Therefore, the lower the OD value, the more the test substance inhibited IgE-derived skin hypersensitivity. Evans blue test results showed that in the group inoculated with 10-fold dilution of mouse serum, there was a significant decrease in all test substance-treated groups except the Lactobacillus confusus PL9001 high-dose group compared to the PBS-administered group. 1. The Lactobacillus confusus PL9001 high-dose group was not significant but decreased compared to the PBS-administered group.

These results indicate that L. coprophilus PL9001 (KCCM-10245), Lactobacillus fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and B. infantis PL9506 (KCCM-10406) have inhibitory effects on skin sensitization. Shows that there is.

Experimental Example 2 Systemic Hypersensitivity Inhibition Test

Systemic hypersensitivity reactions have been reported to increase cytokine associated with IgE. To date, systemic hypersensitivity reactions are known to increase Th2 related cytokine and to increase or decrease Th1 related cytokine mildly. Therefore, in this experiment, the systemic hypersensitivity reactions of Lactobacillus in experimental animals induced systemic hypersensitivity reactions were measured by measuring the changes of cytokine derived from Th1 / Th2 cells.

Materials and methods

Test substance

Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9506 (KCCM-10406) were lyophilized and stored under refrigeration under sealed conditions. Diluted in sterile PBS before administration of the test material was incubated for 1 hour in a 37 ℃ Shaking incubator.

Laboratory animals

50 6-week-old BALB / C mice were obtained from Charles River (Japan). High and low dose groups were set for each test substance, including negative and positive controls, and 5 dogs were placed in each group. 22 ± 3 ℃, relative humidity 50 ± 10%, ventilation times 10-12 times / hr, lighting time 12 hours (08: 00-20: 00), during the whole experimental period in the animal room controlled by the illuminance 150-200 lux The animals were bred, and the water and feed were freely supplied.

Administration of Test Substances in Rats

In the experimental group, strains of Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9506 (KCCM-10406) were treated with 2 × 10 ⁹ CFU / ml for 21 days. And 2 × 10 ⁷ CFU / ml, and orally administered 0.5 ml each time between 10 am and 12 am once a day. Negative and positive controls were administered sterile PBS instead of test material.

Induced systemic anaphylaxis

50 μl Ovalbumin (Sigma: grade V) and 2 mg of 0.2 ml of Al (OH) ₃ were mixed well as an inducer of the hypersensitivity reaction in the entire experimental group and intraperitoneally administered once each on the 0 and 14 days of the experiment. The negative control group was intraperitoneally administered 0.2 ml of PBS. In order to avoid oral administration and duplication of the test substance, administration was performed between 18 and 20 hours.

Culture of Splenocytes for Cytokine Measurement

Animals were euthanized with ether and spleens were removed aseptically on the 21st day of test substance administration. The spleens were chopped with scissors in a flask containing 5 ml RPMI 1640 medium (Sigma), and then suctioned and discharged through a 24 gauge needle, placed in a 15 ml Falcon tube and centrifuged at 1700 rpm for 15 minutes to discard the supernatant. Red blood cells were hemolyzed by incorporating ACK lysis buffer (Tris-NH ₄ Cl) into the precipitate. The precipitate was then washed three times with PBS and then 4 × 10 ⁶ cells / ml in RPMI 1640 complete medium (RPMI 1640 medium containing 10% heat inactivated fetal calf serum, 100 U / ml penicillin, 100 μg / ml streptomycin). Quantitatively, the cells were aliquoted into 12-well culture plates and incubated for 72 hours in a CO ₂ incubator by adding 100 μg / ml of ovalbumin. Negative control cells were also isolated and cultured with ovalbumin. After incubation, the supernatant was collected by centrifugation at 1500 rpm for 5 minutes, and commercialized IL-2, IL-5, IL-12 (Bender Medsystems, Vienna) Elisa kit and IL-4, IL-6, IL-10, TNF- The amount of cytokines expressed using α, INF-γ (Chemicon International, CA, USA) Elisa kit was measured.

Statistical processing

Duncan's multiple test of SAS was used to test group significance.

Results and Discussion

IgE is increased by IL-4 derived from Th2 cells stimulated by IL-1, and IL-5, IL-6 and IL-10 function similar to IL-4 as Th2 cell derived cytokines. IL-10 is derived from Th2 cells, but specifically inhibits cytokine derived from Th1 cells. On the other hand, Th1 cell-derived cytokines inhibit IL-4, reducing the production of IgE, which are IL-2, IL-12 and INF-γ. IL-2 is a cytokine derived from Th1 cell stimulated by IL-12 and induces proliferation of T cell and NK cell and reduces IgE production by inhibiting IL-4.

In the results of FIG. 2A, Il-2 increased little in the positive control group, and significantly increased in the experimental substance-administered group compared with the positive control group. In addition, the high concentration group was significantly increased in each experimental material than the low concentration group. This shows that each test substance has an effect of inhibiting IgE production and preventing systemic hypersensitivity.

IL-4 is a cytokine derived from Th2 cells and is an important factor for producing IgE in B cells, and also plays a role in reducing IL-6 and TNF-α. It also acts to increase the activity of MHC Ⅱ class, APC (antigen presenting cell) and is used as an important indicator of immune response. In FIG. 2B, IL-4 was increased in the positive control group compared to the negative control group, and the test substance-administered group was significantly decreased in comparison with the positive control group. There was no significant difference between the doses, but the high-dose group tended to decrease compared to the low-dose group.

IL-5 is derived from Th2 cells and helps IL-4 to increase IgE synthesis. In FIG. 2C, IL-5 significantly increased in the positive control group compared to the negative control group.

IL-6 is a cytokine derived from Th2 cells, and showed similar results with IL-4. In the low-dose group of Bifidobacterium infantis, there was no significant but decreasing trend compared to the positive control group (FIG. 2D).

IL-10 is derived from Th2 cells and specifically has Th1 cytokine inhibitory function. Thus, similar results to IL-4 and the like were expected, but rather, the cytokine derived from Th1 cells showed a tendency to increase compared to the positive control group (Fig. 2E). Since IL-10 has an inhibitory effect on Th1 cell-derived cytokine, it is thought to be slightly elevated to further suppress the increase of Th1 cell-derived cytokines IL-2, IL-12, INF-γ.

IL-12 is a cytokine derived from Th1 cell and functions to inhibit IgE production by increasing expression of INF-γ and IL-2. In FIG. 2F, only the Lactobacillus confusus PL9001 (KCCM-10245), the low concentration group and the L. fermentum PL9005 (KCCM-10250) high concentration group increased significantly compared to the positive control group, except for the L. plantarum PL9011 (KCCM-10358) low concentration group. There is no significance in the group but it is increasing.

INF-γ is derived from Th1 cell, and when the systemic hypersensitivity reaction was induced, the positive control group showed a tendency to decrease compared to the negative control group. (FIG. 2G).

TNF-α is a cytokine derived from Macrophage, which acts on inflammatory cells and is involved in chronic inflammation. There was no significant change in the negative and positive controls, and the Lactobacillus group except for Bifidobacterium infantis PL9506 (KCCM-10406) was significantly decreased in the high dose group. Bifidobacterium infantis was significantly increased in the low dose group (FIG. 2H). In other words, other lactic acid bacteria except Bifidobacterium infantis PL9506 (KCCM-10406) administration group have been shown to be effective in reducing cytokine.

As shown in the results of Experimental Examples 1 and 2, Lactobacillus copropylus PL9001 (KCCM-10245), Lactobacillus latement PL9005 (KCCM-10250), Lactobacillus plantarum PL9011 (KCCM-10358) and Bifido of the present invention. Bacterial Infantis PL9506 (KCCM-10406) has been shown to have an allergic inhibitory effect in both animal experiments.

As mentioned above, the Lactobacillus copropylus PL9001 (KCCM-10245), Lactobacillus permanent PL9005 (KCCM-10250), Lactobacillus plantarum PL9011 (KCCM-10358) and Bifidobacteria Infantis PL9506 of the present invention. (KCCM-10406) can prevent or treat skin allergies, food allergies and systemic allergies and allergic diseases. In addition, PL strain, PL strain lysate, or PL strain culture filtrate of the present invention can be used for the prevention and treatment, PL strain and PL strain culture filtrate is allergic related diseases including atopic eczema, allergic rhinitis, dermatitis It can be used for the purpose of prevention and treatment. In addition, fermented foods using the strains can be used for the purpose of preventing and treating food allergies.

1. Concentration of Evans blue deposited in rat skin tissue. Negative control group (NST) was orally administered PBS for 21 days, PBS was administered intradermally during serum sensitization. Positive control group (PST) was orally administered with PBS for 21 days, and serum of Anaphylaxis-induced mice was intradermally administered. High concentration group (H) and low concentration group (L) were administered to Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9508 (KCCM-10406) strains. Were administered at 1 × 10 ⁹ CFU / ml and 1 × 10 ⁷ CFU / ml for 21 days, respectively, and the serum of Anaphylaxis-induced mice was intradermally administered. Each letter on the bar graph is significantly different. p <0.05

Figure 2. IL-2 (A), IL-4 (B), IL-5 (C), IL-6 (D), IL-10 (E), IL-12 (F in Spleen Cell Culture Supernatants ), INF-γ (G), TNF-α (H) expression concentrations. Negative control group (NST) was orally administered with 0.5ml of PBS for 21 days, intraperitoneally administered with 0.2ml of PBS on days 0 and 14, and splenocytes were separated on day 21 and incubated with 100 μl / ml OA for 72 hours. The positive control group (PST) was orally administered 0.5 ml of PBS for 21 days, 0.2 ml of OA intraperitoneally on day 0 and 14, and splenocytes were separated on day 21 and incubated with 100 μl / ml OA for 72 hours. High and low concentration groups were administered Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9508 (KCCM-10406). (H) 1 × 10 9 CFU / ml and (L) 1 × 10 7 CFU / ml for 21 days, and spleen cells were isolated on day 21 after OA 0.2ml intraperitoneal administration on day 0 and day 14, and 100 μl / Incubated for 72 hours with ml OA. Each letter on the bar graph is significantly different. p <0.05

Claims (9)

delete delete An allergy preventing or alleviating agent comprising Bifidobacterium infantis Strain PL9506, KCCM-10406. The method of claim 3, wherein the disease is allergic related diseases, including allergic rhinitis, allergic dermatitis, atopic eczema, asthma, food allergy. Cosmetics for persons with atopic skin, comprising Bifidobacterium infantis Strain PL9506, KCCM-10406. delete delete delete delete