TW201223534A - Novel Lactobacillus strains and their uses for modulating immune response - Google Patents

Abstract

The present invention relates to novel Lactobacillus strains, named MP137 and MP108, and their uses for modulating immune response. In particular, the Lactobacillus strains of the invention can promote Th1 immune response and inhibit Th2 immune response.

Description

201223534 6. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to novel lactic acid strains and their use for modulating immune responses. [Prior Art] The immune system is a defense system used by the body to resist foreign microorganisms or toxins. In the immune system, T cells may be differentiated into type 1 helper tau (Thl), type 2 helper T cells (Th2), and regulatory T cells (Treg) after stimulation. Among them, 'Thl cells lead cell-mediated immune response' can secrete interferon-γ (IFN-γ), and promote the proliferation of dendritic cells and macrophages secreting cytokines-12 (IL-12) and cytotoxic killing of tau lymphocytes. Helps fight against viral infections and cancer cells. Th2 cells dominate the humoral immune response, secreting cytokines-4 (IL-4), cytokine _5 (IL-5) and cytokine-13 (IL-13), promoting B cell proliferation and stimulating immunoglobulin e (igE) is produced against free pathogens; however, excessive Th2 responses may cause inflammation and allergic reactions such as asthma, allergic rhinitis, eczema, measles, and gastrointestinal disorders. In addition, s-controlled T cells are negatively regulated by the balance of Th1 and Th2 cells, and can be performed by secreting TGF-β or IL-10. It has been reported that regulated tau cells can inhibit autoimmune and allergic or asthmatic reactions and are currently considered to be useful for the prevention and treatment of allergic diseases. A study has shown that lactic acid bacteria (•"(10)Sp.) have immunomodulatory effects, can inhibit inflammation or alleviate allergic diseases such as atopic dermatitis, allergic rhinitis or asthma. The mechanism of action includes regulating the secretion, control and cytokine secretion.

The balance of Th2 response and the production of antibodies. However, the efficacy of lactic acid bacteria is Stmin-speciflc effects, which varies according to the type of strain, and there are many lactic acid strains in nature that have not yet been discovered or adequately studied. SUMMARY OF THE INVENTION 201223534 Cumming isolates a novel lactic acid strain i (10) from a Taiwanese healthy child's specimen, which is different from the conventionally known strain. The novel strain of the present invention has a plague, and the n-reaction of the nucleus tends to be conditioned, and the immune response should be countered to prevent infection and reduce allergic reactions. Therefore, in one aspect, the present invention provides an isolated lactic acid strain which has a characteristic selected from the group consisting of: Mpi37 If L is deposited in the Food Industry Development Institute of the Republic of China, and is registered. 910484; and the deletion of 8 strains, deposited in the Institute of Food Industry Development of the People's Republic of China, the registration number is BCRC9i〇483. Specifically, the present invention provides a new lactic acid strain _137 and a touch 〇8. In another aspect, the present invention provides a combination comprising the aforementioned lactic acid strain, wherein the composition may be a drug or a food, which may modulate an individual's immune response, and the name 疋 promotes the individual's immune response toward the Th1 immune response, thereby avoiding an immune response. More specifically, it reduces the allergic reaction in an individual. The composition of the present invention has the effect of treating asthma or allergic rhinitis or against intestinal pathogenic infections. Further, the present invention also provides the use of the lactic acid strain described herein for the preparation of a secreted immune response, a medicament for treating asthma or allergic rhinitis or a drug or food for infection with a pathogenic bacteria. The invention also includes a method for modulating an immune response, improving intestinal vaginal immunity, inhibiting an immune response, treating acute asthma or allergic rhinitis, or combating intestinal tract infections in a desired individual, which includes an effective amount of The lactic acid strain is administered to the individual. 'Details of various specific examples of the invention are as follows. The features of the present invention will be apparent from the following detailed description of the specific embodiments. There is no need for further elaboration, and it is believed that those skilled in the art to which the present invention pertains can utilize the invention to the broadest extent based on the foregoing description. Therefore, the description of the understanding of the town is for illustrative purposes only and is not intended to limit the remaining disclosure. 201223534 [Embodiment] All technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains, unless otherwise stated. The article "a" as used herein refers to one or more (i.e., at least one) grammatical acceptance of the article. The present invention provides an isolated lactic acid strain having the characteristics of MP137 or (10) strain of Sichuan, wherein the strain 7 is based on the February 26th, 4th, 8th, and 8th, the registration number is BCRC910484; And the MP108 strain was deposited in the Food Industry Development Research Institute of the Republic of China Foundation on September 6th, 2012. The deposit number is BCRC910483. In particular, the present invention provides a strain of 137 and/or strain. The lactic acid strains MP137 and MP1 〇8 of the present invention were isolated from the source of feces from healthy children in Taiwan. After preliminary tests, these strains are Gram-positive bacilli (Figs. 1 and 2)' and do not have enzymes, oxidase activity and motility, do not produce endospores, and grow under aerobic and anaerobic conditions. Further, 16S rDNA sequence analysis and ap 50 CHL system identification were performed, and the results are shown in Figures 3 and 4 and Table 1 and Table 2 (Example 2). Based on the comprehensive identification results, Μρ137 was identified as a novel strain of Lactobacillus casei sub-species "Fu-__ subsp.", and _108 was confirmed as a novel strain of Lactobacillus rhamnosus (). This hair, MP137 and The MP108 strain may be present in any manner, including live or dead form, as well as equivalent strains having the same characteristics and the cells or products derived from such strains. The term "immune regulation" as used herein, when used When a substance is described, it means that the substance has the function of changing or adjusting at least one immune system, including but not limited to, changing or adjusting the immune system, the member cells or the action molecules (for example, cytokines and antibodies). Quantity (or content) and / or activity. Various test methods have been developed in related fields to evaluate the level adjustment function of substances. As used herein, the term "treatment" includes the purpose of healing, healing, alleviating, improving, or affecting the disease, the symptoms of the disease, the tendency of the disease to cause thieves, or the tendency to develop the disease. The individual will be included or administered or administered to an individual having the disease, the symptoms of the disease, or a predisposition to the disease, or other treatment of the individual. ==^ issued in August, treatment of asthma or allergic rhinitis including the need to give a need to save the u H knife; ^ to reduce or reduce, slow asthma or allergic rhinitis symptoms, such as, reduce lung inflammation and improve nasal symptoms (such as itchy ears, itchy throat, red eyes, tears, etc.). The lactic acid g strain of the present invention has an immunomodulatory effect. In the production of specific f γ, and inhibition of the individual's 1L · 4, IL_5 or its production, for example, enhance the individual two and =: Pan Free Lu Mei i: Those who know the cytokines 1L·12 and Cong-γ negative The balance of TM and Th2 reaction can be used for the prevention of allergic diseases and therefore 'the milk of the invention (four) shoots to promote the immune response of the individual' (4) Th2 free of sputum, contributes to the round of dyeing and falling two: Ming Erding The second knowledge can enhance the individual's important role, and it will be secreted to the intestinal complex. In this way, the lactic acid strain of the present invention can enhance the intestinal immunity of the individual. ,also. Thus, in yet another embodiment, the milk strain of the present invention may be infected, including but not limited to Salmonella and Escherichia coli. In the special ^2012 letter 201223534 ί hair inhibits or replaces the intestinal pathogenic bacteria adsorbed on the intestine, thus "or the efficacy of treatment of intestinal pathogenic bacteria infection. In the case of the resistance of H production ^ lactic acid strain of the present day The lower lungs can be inflamed by lowering the breathing. Therefore, the lactic acid strain of the present invention is a kind of respiratory inflammatory disease, and the bronchial tube is caused by edema of the tube wall, so that the airway resistance is increased, and the sputum is boring, causing sudden death. Standard animal models have been developed in the field to measure 4 respiratory abilities. See the following examples. The secret genus is known to the general knowledge, allergic rhinitis shows a trend of 2, 2: and, in recent years, Thl/Th2 * Epidemic regulator has been the focus of prevention: ίίίί rhinitis - According to the present invention, the milk S text strain described herein can be used to combat allergic rhinitis. Used with other anti-allergic drugs. The anti-allergic drugs described herein can be used in the field to treat allergies, especially for the treatment of allergic rhinitis, including but not antihistamines, de-congestants and Anti-inflammatory drugs (eg, leukotriene antagonist, aldol, mast cell stabilizer, cromolyn, ketotifen, etc.). In a specific example, the lactic acid strain of the present invention is used in combination with an antihistamine. The lactic acid strain of the invention may be used as the anti-allergic drug at the same time or sequentially. On the other hand, the present invention also provides a composition comprising the aforementioned lactic acid strain. In one example, the composition of the present invention comprises the MPU7 strain. In another example, the compositions of the present invention comprise sputum 108. In particular, the compositions of the present invention can be used to modulate an individual's immune response. In one embodiment, the compositions of the present invention modulate the production of cytokines For example, increasing the production of IL-12, IL-10 or IFN-γ in an individual, and inhibiting the production of IL4, IL-5 or IL-13 in an individual. In another embodiment, the composition of the invention may modulate the individual The production of immunoglobulins, for example, enhances the production of IgG2a and inhibits the production of IgE in individuals. Thus, the composition of the present invention can promote the immune response of an individual to a Th1 immune response to inhibit Th2 immunity. Should help to fight against bacterial infections and reduce allergic reactions. 201223534 The composition of the f2 invention can also inhibit the production of airway resistance in individuals and / / allergy ί ί ί '. Available (4) for asthma. The composition of this (4) is also available The composition of the present invention which produces IgA on the surface of the two substances can contain an effective amount of the lactic acid strain, and the dosage of the case is determined according to the experience of the method. Preferably, the composition contains ...! The lactic acid bacteria on the lactic acid bacteria, the lactic acid strain containing 2.5xl G9 _5xlGllcfii, the more specific and versatile carrier can be combined with the present invention: == The active ingredient and the Wei method and the #丨 which are known to be suitable for the injection of the test paste may include, but are not limited to, a topical agent or a diluted sugar, a binder; for example, A:: 2 The present invention: a combination of a suspension liquid, a ritual, a solvent, a glycoside, and a soft and hard gelatin capsule. The composition invented in a special 201223534 tL' ί is in the form of a powder. In another specific implementation, it is too private 'to': and σ is in the form of a capsule. Further, the composition of the present invention is preferably administered orally. The composition of the present invention can be used as a medicine or a food, for example, a lactic acid powder or the like. The composition of the present invention is also - not limited to, an antioxidant, for example, a raw material t, a package of dan!: two s: sweeteners, for example, stevia, sugar substitute, saccharin; Blue, curcumin; and preservatives, for example, sodium sulphur, sulfite, benzoic acid, hexamethylene acid, and the like. # t if in the 'invention of the composition of the present invention, including - or a plurality of and including but not limited to anti-group _, de-congesting agent and anti-inflammatory pelvic seed set, including - or a plurality of first dose The unit, its lactic acid strain of the invention, and one or more second doses; an effective amount of other anti-allergic drugs. Specifically, the lactic acid functional strain can be used simultaneously with the antiallergic drug or sequentially. μ本' also includes a method for modulating an immune response in a desired individual, enhancing an immune response to the intestine, treating asthma or allergic rhinitis, or combating intestinal sensation=sensation τ, which includes the lactic acid S which will be effective* The strain is administered to the individual I. In the r example, the lactic acid strain is a 37 strain. In another example, the lactic acid strain is Cong 08. Preferably, the lactic acid strain is administered in a dose to the desired individual towel. In the method of the present invention, the dose to be administered may be adjusted as needed. Preferably, the lactic acid bacteria is administered at a dose of 108 cfU or more per day. In a specific example, the lactic acid bacteria in a daily dose of lG9-lG12efU

Strain: In another specific example, a dose of 2.5 x 109 -SxloUcfU of the acid-stained strain was taken; in another embodiment, a lactic acid strain of 5 x 109 cfU per day was administered. Those skilled in the art will be able to formulate compositions of the present invention based on the teachings herein, using any of the conventional methods and techniques. 9 201223534 The present invention will be embodied by the following examples, but is limited to the examples set forth in this specification. Example 1 : Separation of strains and collection of stool samples from healthy children in Taiwan, cultured in R〇g〇sa plate medium of (10) selective medium at 37 ° C for 1 to 72 hours to obtain a suspected strain of Lactobacillus. Each colony. The culture was applied to the MRS plate for anaerobic incubation at 3rc for 48 to 72 hours, and the 菌ίΓί plate was used for the purification of the colony, and the isolates were obtained as described in Example 2 to obtain the isolated strain ΜΡ137 and] Ϊ́ρι〇8. The isolated strain was inoculated on the Mrs plate, and after anaerobic culture for 48 to 72 hours, the single-colony colony was picked and inoculated into the fresh pass 8 culture medium until the strain grew well (the turbidity judged by the naked eye), and then 1% was taken. The bacterial solution is transferred to another solution for 18 to 24 hours at a suitable temperature, and this step is repeated to activate the ii body 2. For humans, the resulting bacterial solution is cultured for subsequent testing. Example 2: Strain identification 2.1 Preliminary analysis A preliminary analysis was carried out according to the standard method. The results of the present invention were isolated strains 37 and deleted 08 as Gram-positive bacilli (Figs. 1 and 2), and were not exposed to enzymes, oxidases and exercise. Sexuality does not produce endospore, and it is used in both oxygen and anaerobic conditions. 2.2 16S rDNA PCR analysis The isolated halogen plants MP137 and Mpi〇8 of the present invention were subjected to Mg PCR analysis. The genomic DNA of the strain was extracted using a commercial kit (Jigacterial Gen〇mic DNy! Mimprep Kit, Anxygen Bi〇science). The positive and negative primers were added to the PCR tube (l6S_F: GGAGTTTGATCCTGGCTCAG(SEQIDNO: 1) 16S .R2 : AAGGAGGTGAT CCAGCCGCA ( SEQ ID NO: 2 ) ) . Reagents such as DNA polymerase, buffer, dNTPs, etc., are as follows. Genomic DNA 1-2μί(1〇〇ηδ) 201223534 0.5 pL 10 pL 8pL 1 μΐ 1 μί 77.5 Μΐ 100 μΐ 7^DNA polymerase (TakaraCo.) 1 Ox magnesium flush dNTP mixture (2.5 mM) Forward primer 16S-F (10 (^M) reverse primer 16S-R2 (100pM) h20 total volume

The 16S rDNA PCR reaction conditions include Step 1: 95 ° C, 3 minutes. Step 2: 95T, 30 seconds; 60 ° C '30 seconds' 72 ° C, 45 seconds, 3 cycles &; and Step 3 · 7TC. , 10 minutes' was finally placed at 4t to terminate the reaction. After the end of the PCR reaction, the PCR product was analyzed by agarose gel electrophoresis, and the colloid of the PCR product fragment of the predicted size was cut out to commercialize and Ge^pcR DNA.

The sequenced DNA sequence was assembled by a commercially available computer program (Vector NTI's conflg program, Invitrogen Co.) to obtain the correct DNA sequence, which was then transmitted to NCBI (http://wvmlbi>ninL nih.gov/) On the website, the nucleotide BLAST program was used for the alignment analysis, and the 16S rDNA sequence was selected as the preliminary identification result of the strain with the highest similarity. Figures 3 and 4 show the 16S rDNA partial sequences (SEQ ID NO: 3 and SEQ ID NO: 4) of isolates MP137 and MP108. The comparison showed that the isolate MP108 was closest to Lactobacillus rhamnosus (丨//(10) rhamnosus), Lactobacillus zeae, Lactobacillus casei, Lactobacillus subsp.

The subspecies (subsp. /O/erims) can only be found to be more than 98%, while the isolate MP137 is closest to the Lactobacillus casei subsp. subsp., Lactobacillus faecalis, and corn gluten. Bacillus, Lactobacillus kawaii and Lactobacillus rhamnosus have similarities of more than 98%. 2.3 API 50 CHL system identification 11 201223534 For the isolated strains of gas inventions, delete 37 and 08, 糸 鉴定 ’ 表 表 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 According to the results of the API identification system analysis, in the 49 test items, the isolate strain MP137 was close to the standard strain of Lactobacillus casei sub-chicken subspecies, and there were 43 items (Table 1). Table 1 API identification system analysis results of isolate strain MP137 API 50 CHL MP137 Lactobacillus paracasei subsp. paracasei BCRC 12248T 1. Glycerol - 2 · Erythritol - _ 3. D - arabinose - 4. L-arabinose - _ 5 · D · ribose + + 6. D - xylose - _ 7 · L · xylose - an 8-D-side ginseng (Adonitol) — 9· / 5-methyl-xyloside — 10· D-galactose + + 11 · glucose + + 12· D-fructose + + I3· D-mannose + + 14. L-sorbose + — 15· ϋ 糖_ — 16·Dulcitol — — 17. Inositol 12 201223534 18. D-mannitol + + 19. D-sorbitol + a 20. α-methyl-D-mannosidose — — 21. α-Methyl-D-glucoside — 22. N-acetylglucosamine + + 23. Amygdalin One + 24. Arbutin + + 25·Acetulin + + 26水杨苦+ + 27. D-fiber disaccharide + + 28. D-maltose + + 29. D-lactose + + 30. D-melanose one 31. D-sucrose + one 32. D-algae Sugar + + 33. Inulin + a 34. D-pine Melezitose + + 35. D-Raffinose - 36. Amidon - 37. Liver sugar - 38. Xylitol - 39. Gentobiose + + 40. D-Songose (Turanose) + .+ 41. D-Lesose (Lyxose) I - 42. D-Tagotose + + 43. D-fucose I - 44. L - Fucose - 45. D-arabitol - a 46. L-arabitol - 47. Gluconic acid + 13 201223534

43 48. 2-keto-gluconic acid 49. 5-keto-gluconic acid 49 items in the test In addition, according to the identification system analysis results, in the isolate MPH)8 and the standard strain rhamnosylose Head (10)) BCRC 10940τ is close, and there are 44 items (Table 2). ° Table 2 Analysis of API identification system of isolate MP108 API 50 CHL MP108 Lactobacillus rhamonsus BCRC 10940τ 1. Glycerol — 2. Erythritol — 3. D-arabinose — — 4. L-arabinose — _ 5. D-ribose + + 6. D-xylose _ 7. L-xylose — _ 8. D-side acetaminophen (Adonitol) — — 召–甲Base xyloside - a 10. D-galactose + . + lh D-glucose + + 12. D_ fructose + + 13. D-mannose + + 14. L-sorbose + + 15. L-rham Sugar + + 16. Dulcitol - 17. Inositol - 18. D-mannitol + + 14 201223534 19. D-sorbitol + + 20. α-Methyl-D-mannosidase - — 21. α-mercapto-D-glucoside + + 22. Ν-acetamidine glucosamine + + 23. Amygdalin one + 24. Arbutin + + 25. Esculin + + 26. Salicin + + 27. D-fiber disaccharide + 28. D-maltose + + 29. D-lactose + + 30. D-disaccharide - one 31. D-sucrose + - 32. D-trehalose + + 33. Inulin 1 - 34. D-santriose (Melezitose) + 35. D-Raffinose — 36. Amidon I — 37. Liver sugar — 38. Xylitol — _ 39. Gentobiose + + 40. D-Songose (Turanose) + + 41. D-lysose (Lyxose) — 42. D-Tagotose + + 43. D-fucose — 44. L-fucose — — 45. D-arabitol. — — 46. L-arabitol — _ 47. Gluconic acid + — 48. 2-keto-gluconic acid

S 15 201223534

In summary, the above results g Lactobacillus, and the isolate MP108 of the present invention belongs to the rhamnose isolate MPl37, which is a Lactobacillus casei sub-species. Example 3: Immune sputum 丄1 cell experiment External heat killing sputum sputum 3.1.1 Take the tube and take the 7th, put into the centrifuge, the concentration of the body fluid is ^% read, (10) listen to the bacterial body fluid. Icυ cfb/m was placed at 4 ° C for subsequent experiments. 3.1.2 Purification of CD3+ T cells is slow ΓΓίΪ Blood is about (10) ml, take 25 ml of diluted blood, m water sucrose hexamesamine (Ficoll-Hypaque) away = official 'centrifugation 40 〇 xg, 40 minutes, use density Difference, peripheral blood mononuclear cells were isolated, washed with phosphatase buffer, and the number of cells was counted. Peripheral blood mononuclear white blood cells (pBMCs) isolated from human peripheral blood were added to MACS buffer and CD3+ microbeads at appropriate ratios, allowed to stand at 4 ° C for 15 minutes, and then centrifuged 300 x g in 10-20 ml MACS buffer for 10 minutes. Wash off the excess beads. Finally, re-dissolve the cells with a small amount of MACS buffer and prepare to begin purification. First, the MACS column was wetted with 3 ml of MACS buffer. After the droplets were dried, the cells to be purified were added, and after dripping, the tube was washed with 3 ml of MACS buffer to obtain CD3i CD3+ cells, respectively. The cells were reconstituted with RPMI-1640 medium containing 2% serum, and the cells were frozen in liquid nitrogen by adding 10% DMSO, and the cells of CD3· were used for the culture of dendritic cells. 3. Culture of U human dendritic cells The remaining CD3 cells used for purification of T cells contain 1 〇 ° /. Qiqing's 201223534 RPMI-1640 medium was cultured, and additional goo u/ml recombinant human granulocyte giant cell colony stimulating factor (rhGM-CSF) and 400 U/mr recombinant human IL-4 were used to promote mononuclear spheres. Differentiation into immature dendritic cells. The cells were cultured in a 5% C 2 , 37 ° C incubator for 6 to 7 days, and then these suspension cells were washed from the culture plate to obtain dendritic cells having a purity of about 95%. 3丄4 bacterial sample regulation of human dendritic cell immune function The dendritic cells were collected and the number of cells was counted. Adjust the ratio of cells to dendritic cells and dendritic cells to 1:1, 10:1 or 100:1, and use lipopolysaccharide (LPS) 100 as a positive control group. After 48 hours of culture, the culture medium was collected and immunized with enzyme. The assay (ELISA 'eBiosience commercial reagent) measures the secretion of 1^-10 and IL_12 by dendritic cells. Use other lactic acid bacteria as a control, including adding milk-夂 囷 囷 (), Lactobacillus johnsonii _ 1 (丄·知六似洲) and, "Lactobacillus cerevisiae _2 (Z·知办似洲//-2 The results are expressed as the standard deviation of the mean sample distribution (Mean±SEM), j^O.05 indicates a significant difference, **p<〇〇1 indicates a very significant difference. Paired t check.

Lactobacillus reuteri 4 Lactobacillus johnii 2 NC LPS_ NCj The significance of the sample stimulates the secretion of il-12 by dendritic cells IL-12 (pg/ml) 1:1 10:1 — 100:1 502士283 3402±996** 8310±3276* 79 士30* 1186±509* 6346±3411 3士1 69±50 1888±1048 7±5 43±31 692 318* 8 soil 8 105±70 374 193 0±0 4001士 941** ., compared with NC, if * p < 0.05, ** p < 0.01 represents the system 17 201223534 volume 228 soil; 306 695 ± 293 ** 1171 soil 356 ** 139 ± 195 457 ± 311* 1063±355** 66±93 233±368 795±435* 64±50 101±149 834±381* 37±19 256±148 862±268** 11±1 687 soil 318* Compared with NC, if * p<〇.〇5,** p<0.01 represents IL-10 IL-10 (pg/ml) secreted by dendritic cells. 1:1 10:1 100:1 strain-MP137 MPlos Lactobacillus johnii

Lactobacillus johnii _2 NC

LPS --7___ NC indicates the significance of the culture control group. Table 5: Ratio of IL-12 to IL-10 secretion by dendritic cells stimulated by bacterial samples IL-12/IL-10 Proportion 1:1 10:1 100:1 MP137 2.19 Soil 0.68** 5.18±1.91* 12.07±6.22* MP108 1.01 ±0.38* 2.08 ±0.95* 6.4U2.94* Lactobacillus bulgaricus 0.04±0.02 0.32±0.11* 2.54 Soil 0.81** Lactobacillus johnsonii-1 0.11 士0.09 0.16±0.05** 1.01 0.50* Lactobacillus johnsonii-2 0.07 士0.07 0.32±0.10** 0.50±0.18* NC 0.00 士0.00 LPS 13·71±7.35* NC indicates the culture control group, compared with NC, if *p<〇.〇5 , ** p < 0.01 represents statistical significance. As shown in Tables 3 to 5, the isolates MP137 and MP108 of the present invention stimulated dendritic cells to produce more IL-12 and IL-10, which were shown to contribute to the Th1 response and induce the production of regulatory T cells (Treg), and IL. The ratio of -12/IL-10 is higher than 201223534 咼' indicating that the Thl reaction is stronger than the Treg reaction. 3丄5 bacteria clever to the immune function of 1 run by dendritic cells. Next, it was further confirmed that the secretion of IFN-γ, IL-10 and XL-4 was analyzed by the immune function of dendritic cells affecting T cells. The canine cells were co-stimulated with each strain for 48 hours, and these dendritic cells were stopped by γ-rays to reduce their proliferation ability, and the finely woven 赃4 cells/grooves were adjusted while the number of previously prepared CD3+ T cells was adjusted. For lxlO5 cells / affected dendritic cells, fine tT cells produce Cong·γ strains ~---dumb: γ(ρ^ιι

: The two cells were co-cultured for 48 hours, and the supernatant of the cells was collected, and the secretion of fine, IM and 1L_4 was determined by using the ICP (eBi()si commercial reagent). The stimulation of the lysogen (phytoxin 'ΡΗΑ10 μ8/Πΐ1) was used as a control group, and other = control group 'including Lactobacillus kawaii, Lactobacillus johnsonii, and yoghurt j dry _2. The statistical method is the same as described above. 111 37±16* 27±6** 8±1* 10±4 3 Earth 2 2±1 7±6 2±1 43±21* Wind 212±16〇38±Χ3^ 6±2 7±2 12± 9 ΜΡ137 ΜΡ108 Lactobacillus bulgaricus 4 Lactobacillus johnii _2 NC Τ

DC

T+PHA means Τ fine fetus L..., 丄 丄 丄 ,, DC means dendritic cells, T+PHA stands for hemagglutinin. Compared with NC, if P<_=c_ table does not train her test, T table * only T cell to · -rr - λ 19 201223534 Table 7: Strain-affected dendritic cells stimulate T cells to produce IL-10 IL-10 (pg/ml) strain 1:1 10:1 MP137 43 soil 2* 33±14 MP108 21±3** 16±1** Lactobacillus bulgaricus 8±2 7±1 Lactobacillus johnii-1 9 ± 1 9 soil 1 Lactobacillus johnsonii 8 8 ± 1 8 ± 1 NC 7 ± 2 T 5 ± 1 DC 6 ± 1 T + PHA 34 soil 12 * NC indicates the culture control group, T indicates only T cells, DC indicates dendritic cells, and T+PHA indicates T cells plus phytohemagglutinin. Compared with NC, if * p < 0.05, ** p < 0.01 represents statistical significance. Table 8: Strain-affected dendritic cells stimulate T cell production of IL-4 IL-4 (pg/ml) Strain 1:1 10:1 MP137 1±1 1±1 MP108 1±1 1±1 Calcium lactate Bacillus 2±2 1±1 Lactobacillus johnsonii-3 3±3 5±3 Lactobacillus johnsonii 1 1±1 1 soil 1 NC 2±2 T 4±4 DC 1±1 T+PHA 12 soil 3* NC indicates a culture control group, D indicates only T cells, DC indicates dendritic cells, and T+PHA indicates T cells plus phytohemagglutinin. Compared with NC, if * p < 0.05, ** p < 0.01 20 201223534 represents statistical significance. Affecting dendritic cells to stimulate tau cells to produce 丽-γ and L-1〇 secretion

MP137 MP108 Lactobacillus reuteri 4 Lactobacillus johnii 2 NC T DC T+PHA IFN-y/IL-lO ratio 1:1 10:1 3.76±1.75*1.54±0.31** 1.01±0.24* 1.18±〇.53 0.42±0.28 〇.31±〇.15 0.22±0.13 〇.23±〇.15 7.61 士5.02 2.61±0.94* 0.87±0.36 〇.77±0.26 1.78±1.36 2.39±1.285_ NC_表The non-culture control group 'T indicates only T cells, DC indicates dendritic cells, and T+PHA indicates T cells plus phytohemagglutinin. Compared with NC, if * p < 0.05, ** p < 0.01 represents statistical significance. As shown in Tables 6 to 9, the isolates MH37 and MP108 of the present invention can be visualized, and dendritic cells stimulate T cells to secrete IFN-γ and IL-10, but the expression of IL-4 is lower, indicating that T cells can be promoted to Thl. Reacts and stimulates IL_1 secretion and inhibits Th2 response. 3. 2 Animal experiment 3·2·1 Preparation of bacterial powder After centrifugation of the culture liquid of the test bacteria, the supernatant is removed, leaving the bacterial body ^ 'add appropriate protective agent' to be placed at -8 (TC pre-freeze Overnight, the sample was placed in a buncher and lyophilized in a row to obtain a bacterial powder, including the 137 powder (3.2xl〇ncfU/g) and the Mp1〇8 powder (18xl〇"cfti/g of the present invention. 21 201223534 3.2.2 Animals and camping silver treatment Balb/c female rats were used in the animal room of Taipei Medical University. The mice in the ι〇 group were kept in separate cages and fed freely. Drinking water and Dian will give extra powder samples in a tube-and-tube manner every five days; after six weeks of feeding, they will sacrifice 'to conduct various functional evaluation tests to regulate allergic immune response. According to the ratio of experimental animals to human body surface area The dose was converted into a small tube feeding dose of 2.6xl〇8 Cfu/time/mouse, once a day, this dose ^ is twice the dose, called the 1st dose group, also carried out 〇.5 times the dose group, 丨Dandan group and control group, each dose group is as follows: 11 times 0.5 times dose group: double dose group daily feeding 〇·2 Ml contains 1.3xl08 cfU of bacterial powder (equivalent to adult dose 5χ1〇10); ^ 5 times dose level daily chain 0.2 1111 contains 2.6乂108〇£11 probiotic amount of bacteria (equivalent to adult dose Lxl〇ii); Control group: daily feeding 0.2 1111 containing 1.3\109 pool of probiotics powder (equivalent to adult dose 5xl〇u); and daily feeding by tube, pipe feeding the same volume of 0.2 ml Sterile distilled water. * The ratio of the surface area of the mouse to the human body is 〇.〇〇26,7 kg of body weight of the adult per gram of test substance, equivalent to a dose of 26 grams per day for 20 grams of mice.卩 From the egg to (〇 VA) allergic gas # animal model, every two mice into the Wei cavity injection, given OVA antigen and adjuvant mixed, the reduction will be carried out (four) blood collection, blood centrifuge 12 , _ _, , 钟彳清 'Save in _ 耽 to carry out the subsequent antibody immunization points!! Method for the second reading of the powder samples for six weeks, give the mice inhaled 〇 VA antigen ' ί ^ 彳 sacrifice milk collection lung Wash the spleen and spleen cells, and perform lung washing and spleen cytokine secretion. Figure 5 shows the mouse 5 test procedure. 22 201223534 3·2·4 Body Weight Test During the animal test, the body weight of the mice was recorded every two weeks to estimate the effect of feeding probiotics on the growth of the mice. Table 10 is the record of 'ft' Condition ' to review il〇: mouse weight measurement results ~, σ fruit.

19.31 Soil 0.23 19,12^54 20.15 nmM MP108

0.5 times 18.89 ± 0.33 19.00 ± 0.34 1 time 19.98 ± 0.47 20.17 ± 0.53 - J times 19.93 ± 0.23 20.00 ± 0.45 MP137 20.82 ± ο% 21.65 ± 0.54 21, 42 ± 〇, λ5 22.83 ± 〇 4 〇 23.08 ± 〇 4 〇 ^83 ^36 〇·5 times 19.01 ±0.43 19.42 ± 0.52 21.02 ± η plus

3.2.S Determination of Specific Immunoglobulin Concentration in Blood In the process of establishing a mouse animal model, blood sampling at different time points was performed to measure the anti-potency of IgA, IgE, and IgG2a of anti-〇VA antigen in serum. The determination of the antibody will be carried out by using an ELISA reagent, and the concentration of each antibody will be calculated by measuring the absorbance by an ELISA reader. Tables 13 to 13 show the measurement results. IgA of small-mouse serum-saturated OVA antigen will be measured _ ~~-蔓·^!^ Week 1~3rd week 5th week ± 0.005^ 0.062 ± 0.008 〇j45 Soil 0.046 0.345 ± 0.078 MPl〇8 -- ---" 0.5 times 0.045 ± 0.004 〇113 ± 〇〇11 〇·666 〇146a 〇873 ± 〇〇44b 1 times 0.042 ± 0.001 0.098 ± 0.002 〇.717 soil 0.214b 1.150 ± 0.146b —ϋΙ_0.084 Soil 0.003 0.463 Soil 0.053 0.710 ±0.056 23 201223534 MP137 0.5 times 0-047 ± 0.003 0.089 ± 0.005 0.648 ± 0.1l9a 0.798 ± 0.215a 1 times 0.049 ± 0.004 0.100 ±0.008 0.554 ± 0.101 0.730 ± 0.072 5 times 0.047 ± 0.006 0.086 ± 0.019 0.378 ±0.046 0.6Q7 + n 0S7 a, b and c < 0.001). The table reading control group has statistical significance (10) < () take M < 〇〇 1; c, households can be seen from Table 11 ' ”... after the mice are fed the powder of the isolate of the present invention, body = The amount of specific IgA antibody in the OVA antigen was increased, indicating an effect of improving intestinal immunity. Week 0, Week 3, Week 3, 5th control group, 0.001 ± 0.0005 0.045 ± 0.005 0.057 Soil 0.007 0.348 ± 〇 126 ΜΡ108 一一〇·5 times 0.002 ± 0.0004 0.043 ± 0.005 0.014 ±0.002b 0.082 ± 〇.〇26b 1 times 0 0.030 soil 0.005 0.017 ±0.004b 0.065 ± 〇.〇i7b 5 times 0 0.016 ± 0.006a 0.017 ±0.004 b 0-057 + π n^b 0.5 times 〇_〇〇〇2 ± 0.0002 0.023 ± 0.006 0.021 ± 0.003b 〇·1ΐ5 士& 1 times 0 0.020 ±0.008 0.013 ±〇.〇〇4b 0.128 ±〇 *〇61 ~~~0.0002 d: 0.0002 0.012 ± 0.004b Q.Q14 ± 0.004b Qjq^. 〇^ a, b and c indicate statistically significant (a,

&lt;0.001) 〇. C, P As is apparent from Table 12, the production of a specific IgE antibody against OVA antigen by the mouse in the bacterium of the isolate of the present invention was inhibited by the effect of suppressing the allergic reaction. 'Expression is poor' 1: IgG2a with anti-OVA antigen in mouse serum contains tears, 丨3 weeks 篦 week 1 -&quot;-two two ^ 0.002 soil 0.0005 0.003 ±(λ0007 0.1 MPios &quot; :--- -^ 〇·5 times 0.001 ±0.0005 0.209 soil 0.014a 0.695 ±〇(mb 24 〇6〇" 0.043 201223534 1 1 〇*〇〇2 ± 0.0004 0·180 ±〇.〇28 0.797 ±〇.〇7〇 b 0.69❶士〇.〇56b _〇j〇3± 0.0005 0.192^0.074 0.692 0.697 ±0.066b MP137 &quot;~- 0.5 times 0.002 ±0.0006 0.221 ±0.080a 〇·692±〇.〇73·&gt; 〇6 〇5 〇〇38b 1 times 〇·(8)3±〇._2 0·175±〇.〇41 0.692±0.063b 〇499 soil 〇._a 5 times 50005 〇·182±〇·〇64 0.S78 士〇. 112b i571 ± 0·076«&gt; -————__U^/l ± Ό.υ/Ο = C indicates statistical significance compared with the control group (a, p &lt;0.05; b, p &lt;〇1; c,p As shown in Table 13, after the mice were fed the powder of the isolate of the present invention, the production of the specific IgG24 body in the body of the OVA antigen was significantly increased, indicating that the mouse body was not promoted. The effect of Thl cell immune response. 3·2,6 mouse lung irrigation fluid IL_1 3 Containing Dong determination for 丨®/^ lung lavage fluid to detect IL-13 content in lung lavage fluid by ELISA. The antibody of cytokine is dissolved in buffer and placed at room temperature overnight, every other day.祛* flushing, adding filling buffer to stand at room temperature for 2 hours, lacking = buffer buffer, adding human lung washing solution at room temperature, buffering, adding biotin ship anti- The antibody of cytokines, let stand 2 Xiaoxiao 'Dodge _ liquid rinse, then add the sequel to the room ^ after washing with the stripping buffer', finally add the coloring agent to the color, to read _ the wavelength absorption value Freshness. Table 14 shows the results of the determination. <Intra-IL-13 content, the result ϊίτ- ——____676.2±75.06 MPlos ~~-- 413.8±42.0b 330.6±49.8C 488.4±21.0a 0.5 times ΜΡ137 25 201223534 519.3±46.88 0.5 times 1 times 5 times a, b and c &lt; 0.001) 〇 517.8 ± 16.78 405.2 ± 107.14c means statistically significant compared with the control group (a, p &lt;〇.〇5; b, ρ &lt;0.〇ι;

From the results of Table 14, it was found that the mice significantly reduced the amount of secretion of the IL rinsing liquid IL_13 after the powder of the isolate of the present invention, indicating that the Th2 response was inhibited. The cytokine secretion of the spleen cells of the mice was determined by removing the spleen from the burdock and then preparing the spleen into a single cell suspension. Further, the red spheroid was removed by the blister, and the white blood cells were _ TMSS solution. Cleaning is a step forward. After the isolated cells are adjusted to the appropriate concentration, the lymphocytes are stimulated by the allergens that have been quantified. After IL ^ manufacturing = culture, the upper layer is secreted to determine its cytokines. Table 1 (1) Determination of cytokine secretion in the spleen (legs)

76.8±10.46 MP108 〇·5 times 1 times 5 times — ΜΡ137 59_2±5.71 47.2±7.09a 19.2±4.27c 39.33±7.55a 40.8±12.09 36.0±9.88a --- ____UU*VTy.〇〇&lt;0 f〇 f) The C table has no statistical significance compared with the control group. &lt;0·05; b, P &lt;0.01; It can be seen from Table I5 that 'small gas is fed to the isolate of the isolate of the present invention, spleen 26 201223534 The amount of IL-5 secreted by the visceral cells was significantly decreased, indicating that the sputum response was inhibited. 3·2·8 mice airway resistance change determination mouse health food] VIP108 powder, feeding dose such as 3 2 2 paragraph after six weeks of anesthesia, money age, given through the pipeline ^ j = same concentration gasification B Methahdine's atomization and asthma, and also recorded changes in respiratory resistance. The results of the sputum gas are expressed as relative resistance as follows: Phase T force test force __ force) / (salt salt ^ fruit as shown in Figure 6 (4) 'age physiological saline sensitized mice resistance 11⁄2 inhaling gasification Sewing increase of the 8 bacteria powder, the beer suction channel resistance in the purchase of her food physiological food _ low, 5 times the age of the letter of the mice, the respiratory resistance of the mice showed a small - resistance In addition, the MP137 bacteria powder was used to measure the resistance of the mouse suction channel. The 5 times dose group·· ^ Tian Guan proud 0.2 ml containing 6.5χ1〇6 1 times the dose group 5 times the dose control group: , powder (equivalent to adult dose 2.5 Χΐ〇9);, day, instrument 0.2 ml containing 1.3xl〇7 cfU probiotics powder (equivalent to adult dose 5xl〇9); 2 days ^ feeding (^ (5) containing probiotics with phantoms, , the mushroom powder (equivalent to adult dose 2.5xl 〇 10); and mother day also with tube silver operation, tube dumplings with the same volume 〇. 2ml of sterile distilled water. Age f accumulation of whistle is _26,7 〇対 body (four) adults Daily S substance 'equivalent to 2G grams of mice daily grazing _26 This experiment uses the Buxco system to measure small breaths under non-invasive operation

S 27 201223534 « 3 changes. The resistance changes, computer analysis and according to the data collected from the respiratory tract ^, ', ' of the mouse, the results are obtained by the index (Penh value) is not ^Pen^ value = interval (pause) xpiF / pEF; interval = ( Te_Tr) / Tr (p positive spurt flow rate (peak inspiratory fl0w); PEF: maximum exhalation flow rate (peak exPiratory fl0w rate)). Stimulate the respiratory tract of the mouse first ί ΐ ίί ff 盐水 _ _ _ _ _ followed by the low to high chlorinated ethylene 3 '12 · 5 '25 '5G and 1 GGmg / mL) ' The respiratory physiology changes were recorded, as well as the average penh value. As a result, as shown in Fig. 6(B), the respiratory function index value of the mice in the water control group increased with the dose of I inhaled acetylcholine*. Allergic mice, turtles, sputum, 5 times, 1 time, and 5 times, the value of the slashing value is low, and the surface of the respiratory tract is very small, especially for the respiratory tract caused by 50 and 100 mg/mL. Obstructed. 3.2.9 Analysis of the type of inflammatory cells in the lung lavage fluid The heart 鄕 鄕 t 人 37 37 37 37 37 37 37 37 37 37 37 37 37 37 37 37 37 37 菌 菌 37 菌 对 对 对 对 对 对 对 对 对 对After receiving respiratory allergen stimulation, the small emulsion was sacrificed for lung lavage collection. The lung lavage fluid was centrifuged, and the supernatant was taken out and placed in -2 (TC was stored. The washed out cells were used for cyt〇_e instrument to put the cells on the sputum, and then dried in a sputum, and then washed in a lyophilic solution. Glass; ί 'The cells are counted by microscopy. Judging by variegated silk and cell type 'The cells are divided into four categories: single nucleus (_〇 cyte), lymphocyte (lymphocyte), eosinophilic white blood cell (e〇sin) 〇phil) and neutrophil. Figure 7 shows the results of the analysis. The results showed that in the unsensitized mice (na.ive group), there was no inflammation in the lungs. There is almost no accumulation of eosinophilic white &amp; ball and palliative white ball. In sensitized mice (water control group), there is obvious acidity in the lungs due to the inflammation caused by the addition of cells. The white blood cell and the neutral white ball ^. Comparing the experimental group with the water L scale, it was observed that in the small gas 'Eosinophilic white ball and the neutrophil infiltrating lungs with the powder of the present invention, the 201223534 represents the lung situation. There is a trend of reduction, especially in the 1x dose group with significant ginger Inflammation has improved. ~ ' 3.3 bed test (MP108 combined with anti-tissue Zyrtee therapy) 3.3.1 Observed ethnic observation group for 100 to primary medical clinic, suffering from allergic rhinitis for more than one year and nasal congestion, runny nose The severity of cough or throat itching is moderately above 6-13 years old, but the following conditions are excluded: (1) severe asthma; (2) acute asthma attacks in the past three months; (3) Suffering from acute or chronic sinusitis; (4) having used long-acting antihistamines in the past 1 day; (5) using short-acting anti-nasal or systemic effects in the past 3 days (6) in the past 7 days _ over white three Dilute antagonists; rhyme (7) steroid drugs that have been used intranasally, inhaled or systemically in the past 1 month; ' (8) past nasal congestion in the past 3 days; and (9) past 3 months Anti-allergic probiotics for internal use. 3·3·2 MP108 anti-allergic probiotics wins 108 g powder according to the red method and produces a sac, which is 9 cfu/cap ° 3.3.2 test $1]· The first month of treatment (first 〇·3〇天, the treatment period): one anti-histamine 1 ton per day Zyrtec^-one 1-8 anti-allergic probiotic capsules; Treatment/presentation 2 to 3 months (30 to 90 days, maintenance period): discontinuation of antihistamine, taking a mother-day anti-allergic probiotic capsule. 3.3.3 Assessment item 29 201223534 Patient assessment includes: + Total symptom score, divided into nasal symptoms: runny nose, stuffy nose, itchy nose, sneezing; and non-nasal makeup. Day P a main, Vaughan, ± tears. Divided into 4 etc. Head (4) or ear secrets, money itching, red eyes, sputum = symptoms disappear completely. 1 - Slight (symptoms appear, but do not affect individuals). Home) 2. - Moderate (symptoms are obvious, affecting individuals, but do not affect sleep or life from 3 = severe (interfering with living or sleeping, or even unable to work and rest). (2) Nasal resistance test: left and right. (3) Conscious improvement Degree assessment. Divided into 5 equals = great improvement. 1 = some improvement. 2 - no improvement. 3 = some deterioration. 4 = obvious deterioration. Vortex in the 30th, 60th and 90th days of the above (丨) full Resistance test for eight nasal passages, 6th and 9th in the basin (King symptom number and (2 degree evaluation.) 〒 60 60 and 90 days extra 仃 (3) consciously improve China 3.3.4 test results ^Γ00 children participated, and finally a total of 59 completed 3 months test 16:

201223534 Diagnosis has more than one year of allergicity 59 100 history of rhinitis asthma 16 27.12 atopic dermatitis 11 18.64 Table 17: improvement in the overall symptom score improvement number of nasal symptoms 30th day 37% 48 day 60 33% 44 day 90 39% 45 Non-nasal symptoms Day 30 47% 44 Day 60 44% 42 Day 90 48% 40 Nasal + Non-nasal Day 30 4.1% 50 Day 60 37% 48 Day 90 43% 48 Table 17 showed that in 59 children who were included in the analysis, the nasal symptom score decreased by 39%, the non-nasal symptom score decreased by 48%, and the total symptom score (nasal + non-nasal) decreased by 43%. . Table 18: Nasal resistance score _ improvement range _ improvement number left nasal resistance 30th day 8% 32 60th day 26% 42 31 201223534 Day 90—__34%____46 Right nasal resistance 30th day 24% 39 60 Day 30% 42 Day 90 ___45%____46 Average nasal resistance on day 30 16% 38 Day 60 28% 43 Days again - 39% _____ Table 18 shows that in 59 children included in the analysis 'Participate in this trial III After a month, the average nasal resistance improved by nearly 40%. Example 4: Assay analysis against intestinal pathogenic infections This example measures the efficacy of the isolated strains MP108 and MP137 of the present invention against intestinal pathogenic infections by exclusion and displacement experiments. The intestinal pathogens tested in this example include Salmonella subsp. quinquefolium, BCRC10744), and Escherichia coli (co//, BCRC15372), and commercially available lactic acid strains (Lactobacillus casei variety r/zizwwosm, Lcr35 and

Lactobacillus acidophilus, as a scorpion. In the exclusion experiment, the intestinal cell line Caco-2 cells (BCRC60182) were cultured and stabilized in the medium (Dulbecco's modified Eagle, s Medium, DMEM), and then the lactic acid bacteria to be tested were added, wherein the ratio of lactic acid bacteria to Caco 2 cells was 1〇. 1 (1〇MOI, multiplicity of infection). After co-cultivation at 37 ° C for 1.5 hours, the unattached milk was washed away with phosphate buffer (pBS). Next, pathogenic bacteria were added to infect Cac〇_2 cells, and the ratio of the two was pathogenic: Cac0-2 cells were 10:1. Further, 1.5 small pieces were co-cultured at 37 ° C, and the unattached strain was washed with PBS, and Gram stain was performed to count the number of pathogenic bacteria attached to Caco-2 cells. Figure 8 shows the results of the exclusion experiment. (A) Experimental results for Salmonella, and experimental results for the large intestine 32 201223534. Further, in the replacement experiment, the culture conditions were the same as those in the above exclusion experiment, but the pathogenic bacteria were added to the intestinal cell line Caco2 cells (the ratio is pathogenic bacteria: Caco-2 cells = 10:1) at 37. (: After co-cultivation for 15 hours, wash the unattached pathogens with PBS; then add lactic acid bacteria (the ratio is lactic acid bacteria: Caco-2 cells = 10:1), and continue to cultivate 15 at 37t: After washing the unattached strain with PBS, Gram staining was performed to count the number of pathogenic bacteria attached to Caco-2 cells. Figure 9 shows the results of the displacement experiment, in which (A) results for Salmonella were tested. , and (B) is the experimental result for E. coli. The symbol "#" indicates that there is a significant difference between the corpse and the 对照组5, and the symbol "*" indicates that the two groups have significant differences (&lt;5) As shown in Figures 8 and 9, the isolated strains (10) and (7) of the present invention can successfully inhibit or replace the intestinal pathogenic bacteria adsorbed on the intestinal tract, thereby having the effect of fighting pathogenic bacteria infection. ^ As confirmed by the above results, The invention divides and deletes (10) and deletes 37 to immunize f energy, which can promote TM reaction, inhibit Th2 reaction, reduce allergy, hair growth, sexual cell reaction, improve intestinal immunity, and reduce the resistance of asthmatic individuals. The effect is also resistant to intestinal pathogenic bacteria The function of the skilled person will be able to do without departing from the spirit of the invention; the scope of the invention is covered by other disclosures within the scope of the application. [Simple description of the schema] * The previous text and the real money can be borrowed _ Add a pattern

What is the St. 1 Description of the Invention 'Appropriate embodiment a. It should be noted that the present invention is not limited to the descriptions set forth herein.

Separation of strains observed by the strain of Cong 37 under the microscope. The isolated strain was observed under the microscope. ID N,. 3) The 16S ΦΝΑ partial sequence of the isolate 7 (SEQ 33 201223534 Figure 4 is the 16S rDNA partial sequence of the isolate MP108 of the present invention (SEQ ID NO: 4). Figure 5 shows the mouse test procedure of Example 3.2. Fig. 6 shows the results of measurement of changes in respiratory resistance of mice treated with (A) MP108 strain and (B) Mp137 sound of the present invention. ~ 丨η Figure 7 shows analysis of inflammatory cell types in lung lavage fluid of mice 妗 indicates monocytes Eos means acid white blood cells, Neu means: fruit, Mo ball and Lym means lymphocyte cells. Not neutral white Figure 8 shows the results of the exclusion experiment of Example 4, (the experimental results of A bacteria, and (B) are for Escherichia coli ^ for Salmonella Figure 9 shows the results of the displacement experiment of Example 4, the results of a (,, σ fruit. bacteria, and (8) is for the large intestines Shamen 201223534 Sequence Listing &lt;110&gt; Taiwan Toyo Pharmaceutical Industry Co., Ltd. Ltd. Food Industry Development Research Institute &lt;120&gt; Novel lactic acid strain and its use for regulating immune response

&lt;130> IT0163/TY0008TW &lt;150&gt; TW099142822 &lt;151&gt; 2010-12-08 &lt;160> 4 &lt;170&gt; Patentln version 3.5 &lt;210> 1 &lt;211&gt; 20 &lt;212&gt; DNA &lt;213&gt;Artificialsequence&lt;220&gt;&lt;223&gt;Introduction&lt;400&gt; 1 ggagtttgat cctggctcag &lt;210&gt; 2 &lt;211&gt; 20 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt;&lt;400&gt; 2 aaggaggtgat ccagccgca &lt;210&gt; 3 &lt;211&gt; 500 &lt;212&gt;DNA_&lt;213&gt; Lactobacillus paracasei subsp. paracasei &lt;400&gt; 3 tcgaagcaac gcgaagaacc ttaccaggtc ttgacatctt ttgatcacct gagagatcag gtttcccctt cgggggcaaa atgacaggtg gtgtatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttatga ctagttgcca gcatttagtt gggcactcta gtaagactgc cggtgacaaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg gatggtacaa cgagttgcga gaccgcgagg tcaagctaat ctcttaaagc cattctcagt tcggactgta ggctgcaact cgcctacacg aagtcggaat cgctagtaat cgcggatcag cacgccgcgg tgaatacgtt ccc Gggcctt gtacacaccg cccgtcacac catgagagtt tgtaacaccc gaagccggtg gcgtaaccct tttagggagc

&Lt; 210> 4 &lt; 211 &gt; 500 &lt; 212 &gt; DNA &lt; 213 &gt; sugar lactobacilli (Lactobacillus rhamnosus) &lt; 400> 4 tggagtttga tcctggctca ggatgaacgc tggcggcgtg cctaatacat gcaagtcgaa cgagttctcg ttgatgatcg gtgcttgcac cgagattcaa catggaacga gtggcggacg ggtgagtaac cctgccctta agtgggggat aacatttgga aacagatgct of acgtgggtaa 1 201223534 aataccgcga tagatccaag aaccgcatgg ttcttggctg aaagatggcg taagctatcg 240 cttttggatg gacccgcggc gtattagcta gttggtgagg taatggctca ccaaggcgat 300 gatacgtagc cgaactgaga ggttgatcgg ccacattggg actgagacac ggcccaaact 360 cctacgggag gcagcagtag ggaatcttcc acaatggacg caagtctgat ggagcaacgc 420 cgcgtgagtg aagaaggctt tcgggtcgta aaactctgtt gttggagaag aatggtcggc 480 agagtaactg ttgtcggcgt 500 page 2

Claims (1)

201223534 VII. Patent application scope: The lactic acid isolated from the fine H, wherein the milk _ strain has the characteristics selected from the following ft 囷 strain: the 37 strain is deleted, deposited in the Republic of China Corporation and the Institute of Industrial Development, the registration number For bcrc, it is deposited in the Chinese People's Republic of China Foods Development Institute, the registration number is BCRC910483. According to the data? Please apply the lactic acid strain of the first item of the patent scope, in which the lactic acid strain =, deposited in the Food Industry Development Research Institute of the Republic of China, and the registered version of the tiger is BCRC910484. According to the lactic acid strain of the first item of the patent scope, the lactic acid strain is MP108 miscellaneous, and is deposited in the Food and Drug Administration of the Republic of China, and is registered as BCRC91G483.展研九 4. A composition comprising a lactic acid strain as claimed in claims 1 to 3. 5^If the towel is please, please use the composition of the fourth item, which is 魏 囷 or dead 囷. 6. The composition of claim 4, which is in the form of a powder or a capsule. 7. A composition as claimed in claim 4 for use in regulating an individual epidemic response. 8. A composition as claimed in claim 4 for use in regulating the production of a cytokine in an individual. ^ 9. The composition of claim 8 for use in promoting the production of IL-12 or IFN-γ in an individual. 10. The composition of claim 8 for use in boosting the production of IL-10 in an individual. — 11. The composition of claim 8 for use in inhibiting the production of IL-4, IL-5 or IL_B. 12. The composition of claim 4, which is used to modulate the production of immunoglobulin (Ig). 201223534 of P1U body 13. The composition of claim 12, which is used to enhance the production of IgG2a in an individual. 14. The composition of claim 12, which is used to increase the production of IgA in an individual. 15. The composition of claim 12, which is for use in inhibiting the production of IgE in an individual. 16. The composition of claim 4, which is used to enhance an individual's intestinal immunity. 17. A composition as claimed in claim IA, for use in inhibiting an allergic reaction in an individual. 18. The composition of claim 4, for use in the treatment of asthma or allergic rhinitis. 19. The composition of claim 4, for use in combating intestinal pathogen infection. 20. The composition of claim 19, wherein the intestinal pathogen is Salmonella or Escherichia coli. 21. The composition of claim 4, wherein it is a composition. 22. The composition of claim 4, which is in oral form. A pharmaceutical or food for preparing a lactic acid strain according to any one of claims 1 to 3 for the preparation of a sputum immune response, treating asthma or allergic rhinitis or combating intestinal pathogenic infections. Use.