BR122024010955A2 - LACTOBACILLUS PARACASEI STRAINS, FOOD, FOOD ADDITIVE AND PHARMACEUTICAL COMPOSITIONS, AS WELL AS USES OF A COMPOSITION COMPRISING THE STRAINS FOR PREVENTING OR TREATING INTESTINAL DISEASES, ALLERGIC DISEASES, AUTOIMMUNE DISEASES OR INFLAMMATORY DISEASES - Google Patents

Abstract

“LACTOBACILLUS PARACASEI STRAINS, FOOD, FOOD ADDITIVE AND PHARMACEUTICAL COMPOSITIONS, AS WELL AS USES OF A COMPOSITION COMPRISING THE STRAINS FOR PREVENTING OR TREATING BOWEL DISEASES, ALLERGIC DISEASES, AUTOIMMUNE DISEASES OR INFLAMMATORY DISEASES” The present invention pertains to: a novel human-derived probiotic strain that is Lactobacillus paracasei KBL382, Lactobacillus paracasei KBL384 or Lactobacillus paracasei KBL385; and a use of the strain for improving gut health and treating or preventing bowel diseases. The strain according to the present invention has excellent anti-inflammatory and immunomodulatory functions, has an excellent strengthening effect on the tight junctions of the intestinal tract wall, suppresses enteritis-induced weight loss and colon length reduction, thereby exhibiting a therapeutic effect on enteritis and significantly alleviating the symptoms of atopic dermatitis. Therefore, the strain may be useful as a probiotic material to enhance anti-inflammatory effects, strengthen immunity, improve intestinal health functions and alleviate allergic diseases.

Description

[001]Divided from BR 11 2020 022749 8, filed on 09/05/2019. Field of Technique

Field of Technique

[002] The present invention relates to strains of Lactobacillus paracasei and the use thereof. More specifically, the present invention relates to a healthy functional food composition for the relief of allergic symptoms, improvement of immunoregulation, relief of inflammatory symptoms, relief of atopic dermatitis and improvement of intestinal health; and a pharmaceutical composition for the treatment or prevention of allergic diseases, autoimmune diseases, inflammatory diseases, atopic dermatitis and/or intestinal diseases, comprising an effective amount of at least one selected from the group consisting of a KBL382 strain of Lactobacillus paracasei, a KBL384 strain of Lactobacillus paracasei, a KBL385 strain of Lactobacillus paracasei, microbial cells of said strain, cultures of said strain, lysates of said strain and extracts of said strain.

Fundamentals of the Technique

[003] Probiotics refer to microorganisms and their products having antibacterial activities and enzymatic activities to help balance intestinal microorganisms. In addition, probiotics are also defined as live bacteria in the form of a single strain or multiple strains to improve intestinal flora when fed to humans or animals in the form of dried cells or fermentation products. Probiotics must inhabit the human intestine, be non-pathogenic and non-toxic, and survive long enough until they reach the intestine. In addition, probiotics must maintain viability and activity until they are consumed in the released food, be sensitive to antibiotics used to prevent infection, and must not have antibiotic-resistant plasmids. Also, probiotics must be resistant to acids, enzymes, and bile in the intestinal environment.

[004]These probiotics may include, for example, Bacillus sp. which has an excellent ability to produce digestive enzymes such as amylase, protease, lipase, cellulase and phosphatase, Lactobacillus sp. which produces lactic acid and photosynthetic bacteria which prevent bad odor by using the odor-causing substances (such as ammonia, hydrogen sulfide and amines) remaining in the feces of farm animals in a metabolic process.

[005]In particular, Bacillus sp. and Lactobacillus sp. are known as very useful probiotics because they include strains that produce several antibacterial substances. These lactic acid bacteria produce antimicrobial peptides called Bacteriocins, which have antibacterial mechanisms that are not related to the antibiotic resistance mechanism. Bacteriocins have polymorphic characteristics so that their molecular weights, biochemical properties and antimicrobial ranges and mechanisms for the host vary considerably. Klaenhammer defines bacteriocin as a protein or protein complex that has direct antibacterial activity against species related to bacteriocin-producing bacteria.

[006] Meanwhile, irritable bowel syndrome (IBS) is a symptom characterized by abdominal pain and/or irritation associated with altered bowel movement or bowel habits, such symptoms cannot be explained with anatomical or biochemical abnormality. Common symptoms of IBS also include urinary urgency, abdominal distension, and sensation of incomplete bowel movement. Consequently, IBS can be classified as functional gastrointestinal disorders comprising conditions such as functional abdominal distension, non-cardiac chest pain, non-ulcer dyspepsia, and chronic constipation or diarrhea. In particular, in the case of IBS, since the related symptoms affect both the well-being and the normal functional aspect of patients, the disease has a great impact on morbidity and quality of life, in addition to abdominal pain and discomfort.

[007] Inflammatory bowel disease (IBD) is a condition in which chronic abnormal inflammation in the intestine recurs with remission and remission, encompassing all inflammatory bowel diseases, such as Crohn's disease, ulcerative colitis or Behcet's disease, but not limited to them. Much research has been conducted in the field of drug development to treat IBS and IBD. In this regard, several antidepressants are commonly used, although their efficacy in clinical trials is moderate and their clinical usefulness is limited due to significant side effects. Serotonergic medications have also been shown to be effective against global symptoms of IBS. However, the application of these medications has been restricted in various ways due to several recent safety concerns. Consequently, there is increasing interest in the development of a new therapeutic agent for IBS.

[008]Meanwhile, allergy is a biochemical phenomenon that exhibits a unique and altered response to a foreign substance (antigen, allergen). The foreign substance that causes the symptoms is called allergen, while the diseases arising from these symptoms are called allergic diseases. Allergy is a pathological process in the living organism resulting from the antigen-antibody reaction. In general, there are four types of allergies depending on the period until triggering the reaction and the involvement of the complement. Type 1, among these, is the anaphylactic type (immediate type) in which target organs are mainly digestive organs, skin and lungs and common symptoms include gastrointestinal allergy, urticaria, atrophodermatitis, allergic rhinitis and bronchial asthma, etc. The pathological mechanism of Type 1 is known as follows: when antigens contact IgE antibodies bound to the surface of mast cells and basophilic leukocytes, the target cells are activated to secrete chemical transmitters such as histamine, leukotriene and PAF, and then blood vessels and smooth muscles are contracted. Such a mechanism can often be combined with Type 4 (delayed type). In other words, such anaphylaxis and allergic reaction may arise due to a variety of changes in mast cells, etc. The activation of mast cells, which leads to degranulation, is caused by the binding of antigen, anti-IgE, lectin, etc. to Fc receptors, stimulation of anaphylatoxin, etc. or other drugs such as calcium ionophore, 48/80 compound, codeine and synthetic adrenocorticotropic hormone.

[009]Mast cells and basophils in the blood are known as the main cells in the body, causing many allergic diseases such as allergic rhinitis, allergic dermatitis, asthma, food allergy and anaphylactic shock. These cells have receptors (FcRI) for IgE on their surfaces which is an allergy causing antibody and the cells are stimulated by the allergy causing substances (antigen, allergen) to secrete their own allergy causing substances outside the cells (Kim K et al, Eur J Pharmacol, 581:191 - 203, 2008).

[010] Among allergic diseases, atopic dermatitis, as widely known to the public, is a chronic recurrent skin disease that affects newborns or children and may persist into adulthood. Similar to asthma or allergic rhinitis, atopic dermatitis is an inflammatory skin disease associated with local infiltration of T lymphocytes that produce IL-4 and IL-5. IL-4 controls the development of the T helper 2 (Th2) phenotype, resulting in overproduction of immunoglobulins (Ig) and eosinophilia and increased serum IgE levels. 80 to 90% of individuals who tested positive for skin testing to food and inhalant allergens had atopic dermatitis.

[011]There are different treatments to treat or prevent allergic diseases including atopic dermatitis, but no effective treatment has been discovered yet. Some drug-based treatments are known, but even short-term administration of the drug for treatment would develop tolerance and long-term administration could cause serious side effects, and thus, such drug-based treatments of allergic diseases have been avoided recently. Under the circumstances, without the treatment having any obvious and absolute effect, irritating symptoms such as itching and redness of the skin other than allergy often do not improve.

[012] WO 96/29083 and EP 554418 disclose two types of Lactobacillus strains that form colonies in the intestine, i.e. Lactobacillus plantarum 299v (DSM 6595) and Lactobacillus casei ssp. rhamnosus 271 (DSM 6594), etc. EP 415941 discloses a method for preparing a nutrient composition, comprising treating oatmeal porridge with enzymes before mixing it with lactobacilli. US Patent No. 7195906 discloses a strain of Bifidobacterium isolated from the excised and washed human gastrointestinal tract for the treatment of inflammatory diseases, especially gastrointestinal inflammation such as IBD and IBS.

[013]However, no strain having excellent effects on improving gut health, for example, treating IBD and IBS and relieving allergy symptoms has been found yet and in order to find strains having such effects, many research institutions are working on it.

[014]Under the circumstances, the present inventors screened a variety of strains based on the fact that a health-improving effect of probiotics is not a genus- and species-general attribute but rather a strain-specific one (Report of a joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food, London Ontario, Canada, 2002) and identified a new strain having excellent effects on immunoregulation, anti-inflammation activities, allergy relief, improvement of intestinal health and treatment of intestinal diseases. Then, the present invention was completed by confirming the superior effects of these strains.

Summary of the Invention

[015]The purpose of the present invention is to provide a new strain useful for improving intestinal health, treating or preventing intestinal diseases, improving immunoregulation, treating or preventing autoimmune diseases, alleviating allergic symptoms, alleviating inflammatory symptoms, alleviating and treating atopic dermatitis.

[016] In order to achieve the purpose, the present invention provides a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP).

[017] Also, the present invention provides a food composition or food additive composition comprising an effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[018]The present invention also provides a pharmaceutical composition for the treatment or prevention of intestinal diseases, comprising an effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain.

[019]The present invention also provides a pharmaceutical composition for the treatment or prevention of allergic diseases, comprising an effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain.

[020]The present invention also provides a pharmaceutical composition for the treatment or prevention of autoimmune diseases or inflammatory diseases, comprising an effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP), a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[021]The present invention also provides a method for treating intestinal diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain to a subject in need thereof.

[022]The present invention also provides a method for treating allergic diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain to a subject in need thereof.

[023]The present invention also provides a method for treating autoimmune diseases or inflammatory diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP), a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain, and extracts of said strain to a subject in need thereof.

[024]The present invention also provides a composition for the use of preventing or treating intestinal diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[025]The present invention also provides a composition for the use of preventing or treating allergic diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[026]The present invention also provides a composition for the use of preventing or treating autoimmune diseases or inflammatory diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[027] The present invention also provides the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP), a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for intestinal diseases.

[028] The present invention also provides the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for allergic diseases.

[029] The present invention also provides the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for autoimmune diseases or inflammatory diseases.

Brief Description of the Drawings

[030] FIG. 1 illustrates the result confirming the effect of controlling inflammatory cytokines by a KBL382 strain, a KBL384 strain and a KBL385 strain of the present invention in PBMC.

[031] FIG. 2 illustrates the result confirming the effect of controlling gene expression of T cell differentiation markers by a KBL382 strain, a KBL384 strain and a KBL385 strain of the present invention in PBMC.

[032] FIG. 3 illustrates the result of observing the change in the expression level of cytokines, (A) IL-2, (B) IFN-Y, (C) IL-4, (D) IL-13, (E) IL-17A, and (F) IL-10, according to the administration of a KBL382 strain, a KBL384 strain, and a KBL385 strain of the present invention, into PBMC cell lines so that T cells are activated by addition of anti-CD3 antibody.

[033] FIG. 4 illustrates the result confirming the effects on alleviating enteritis through (A) the change in body weight, (B) the change in colon length, (C) the change in colon length and change in cecum weight, (D) H&E staining of colon tissues, and (E) the change in MPO (myeloperoxidase) level in colon tissues after administering each of a KBL382 strain, a KBL384 strain, and a KBL385 strain of the present invention to mouse models that enteritis was induced.

[034] FIG. 5 illustrates the result of observing the effect of strengthening the tight junctions of a KBL382 strain and a KBL385 strain of the present invention through changing the expression level of the genes related to the tight junction of the intestinal tract wall, (A) ZO1, (B) Claudin3, and (C) MUC4.

[035] FIG. 6 illustrates the result of comparing the effects of (A) recovering colon length and (B) improving body weight loss by a KBL382 strain of the present invention and infliximab, an antibody for treating enteritis, which is commercially available, so as to confirm the effect of alleviating enteritis by the KBL382 strain of the present invention.

[036] FIG. 7 illustrates the observation result of (A) the measurement of dermatitis score, (B) the itch-relieving effect, (C) the skin thickness-reducing effect, and (D) the blood IgE concentration-reducing effect by administering a KBL382 strain of the present invention to animal models in which atopic dermatitis was induced, so as to confirm the effect of alleviating atopic symptoms by the KBL382 strain.

Detailed Description of the Invention and Best Modes for Carrying Out the Invention

[037]Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as understood by one skilled in the art. In general, the nomenclature used in this specification is well known in the art and is commonly used.

[038] The present invention confirmed an immunoregulatory effect of microorganisms derived from the human body, and discovered strains of Lactobacillus paracasei having an excellent immunoregulatory effect, namely, KBL382 (Accession No. KCTC13509BP), KBL384 (Accession No. KCTC13510BP) and strain KBL385 (Accession No. KCTC13511BP). Analysis of 16s rDNA of said strain demonstrates that said strain is a new strain that has never been known to the public.

[039]In one embodiment, the present invention relates to a novel probiotic strain of Lactobacillus paracasei KBL382, Lactobacillus paracasei KBL384 or Lactobacillus paracasei KBL385 and said strains are characterized by comprising 16s rDNA sequences of SEQ ID NOs: 1 to 3, respectively, as below.

[040]<SEQ ID NO: 1> 16s rDNA sequence from a KBL382 strain of Lactobacillus paracasei >KBL382 GCAGGTGGCGGGTGCTATACATGCAGTCGACGAGTTCTCGTTGATGATCGGTGCTTGCACC GAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAA GTGGGGGATAACAiπGGAAACAGATGCTAATACC GCATAGATCCAAGAACCGCATGGTTC TTGGCrGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCrAGTTG GTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCAC ATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAA TGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAAC TCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCA GAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATC CGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTCG GCTTA ACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGA ACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGC TGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGAπAGATACC CTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTπCCGCCCTTCAGTGC CGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAG GAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAG AACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGnTCCCCTTCGGGGGCA AAAATGACAGGTGGTGCATGGπGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCG CAA CGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGC CGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGG GCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAAT CTCTTAAAGCCATICTCAGTTCGGACTGTAGGCTGCAACTCGCCTAGACGAAGTCGGAATC GCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGπCCCGGGCCπGTACACACCGC CCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTAGGGAGCGA GCGTCTAAGTGGCTCACGCCT

[041]<SEQ ID NO: 2> 16s rDNA sequence from a KBL384 strain of Lactobacillus paracasei >KBL384 GCCAGTGGGGGGTGCTATACATGCAGTCGAACGAGπCTCGπGATGATCGGTGCπGC ACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCT TAAGTGGGGGATAACATπGGAAACAGATGCT AATACCGCATAGATCCAAGAACCGCATG GTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTA GTTGGTGAGGTAATGGCTCACCCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGG CCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCπCC ACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTπCGGGTCGTA AAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCA ACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGT TATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCC CTCGGC ΠAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGT GGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGC GGCrGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGA TACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGπGGAGGGTTTCCGCCCTTCA GTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTC AAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGiπAATTCGAAGCAACGC GAAGAACCTTACCAGGTαTGACATCπTTGATCACCTGAGAGATCAGGπTCCCCπCGGG GGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGG GTTAAGT CCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGA CTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGAC CTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGC TAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGA ATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACC GCCCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTπAGGGAGC GAGCCGTCTAAGGTGAACCAAAGΠTG

[042]<SEQ ID NO: 3> 16s rDNA sequence from a KBL385 strain of Lactobacillus paracasei >KBL385 GCAGTTGGGGGGAGCTATACATGCAGTCGACGAGTTCTCGTTGATGATCGGTGCTTGCAC CGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTA AGTGGGGGATAACATTTGGAAACAGATGCTAATACCGC ATAGATCCAAGAACCGCATGGTT CTTGGCTGAAAGATGGCGTAAGGTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTT GGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCA CATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACA ATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAA CTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGπGCCGGCGTGACGGTATCCAACC AGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTAT CCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTC TAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGG AACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGG CTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATAC CCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGΠTCCGCCCΠCAGTG CCGCAGCTAACGCAπAAGCATTCCGCCTGGGGAGTACGACCGCAAGGπGAAACTCAAA GGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAA GAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGTTTCCCCTTCGGGGGC AAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGπGGGπAAGTC CC GCAACGAGCGCAACCCπATGACTAGπGCCAGCATTTAGπGGGCACTCTAGTAAGACrG CCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCπATGACCTG GGCTACACACGTGCTACAATGGATGTACAACGAGπGCGAGACCGCGAGGTCAAGCTAA TCTCTTAAAGCCAΠCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATC GCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGC CCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCrπAGGGAGCGA GCCGTCTAAGTGTACAAAGTT

[043] The present invention also confirmed that a KBL382 strain, a KBL384 strain and a KBL385 strain all have a strengthening effect on the tight junction of the intestinal tract wall in addition to anti-inflammatory and immunoregulatory effects, and that administration of a KBL382 strain, a KBL384 strain and a KBL385 strain to animal models in which enteritis was induced markedly alleviated the loss of body weight and the reduction of colon length due to enteritis. Furthermore, it also confirmed that administration of the KBL382 strain, among the above strains, to animal models in which atopic dermatitis was induced markedly improved the dermatitis score, alleviated the symptoms of itching, decreased the skin thickness and markedly improved the blood IgE concentration due to the allergic reaction.

[044] Accordingly, in another embodiment, the present invention relates to a food composition or food additive composition comprising an effective amount of at least one selected from the group consisting of microbial cells of a KBL382 strain, a KBL384 strain or a KBL385 strain, cultures of said strain, lysates of said strain and extracts of said strain.

[045] Said food composition or food additive composition can be readily used as the effective food for improving intestinal health and preventing intestinal diseases, for example, as main ingredients or secondary ingredients of food, food additives, healthy functional food composition or functional beverages, but is not limited to these.

[046] Said food composition or food additive composition can be readily used as the effective food for the relief of allergic symptoms, for example, as main ingredients or secondary ingredients of food, food additives, healthy functional food composition or functional beverages, but is not limited to these.

[047]Furthermore, said food composition or food additive composition can be readily used as the effective food for alleviating autoimmune diseases or inflammatory diseases, for example, as main ingredients or secondary ingredients of food, food additives, healthy functional food composition or functional beverages, but not limited to these.

[048]The term “food” refers to a natural or artificial product comprising at least one nutrient and more preferably, refers to a product that becomes edible through certain processing, usually encompassing all foods, food additives, health functional foods and functional beverages.

[049]The food that may comprise said food composition according to the present invention as an additive may include, for example, different types of food, beverages, chewing gum, tea, vitamin complex and functional food. Further, the food of the present invention includes special nutritional food (e.g., modified milk, baby food), processed meat products, fish meat products, tofu, muk, noodles (e.g., ramen, Asian noodles), bakery products, health food supplement, seasoning products (e.g., soy sauce, soybean paste, red pepper paste, mixed paste), sauces, confectionery (e.g., snacks), candy, chocolate, chewing gum, ice cream, dairy products (e.g., fermented milk, cheese), other processed food, Kim-chi, salty food (e.g., different kinds of Kim-chi, pickled food), beverages (e.g., fruit juice, vegetable juice, soy milk, fermented beverages), and natural seasonings (e.g., bouillon powder for ramen), but is not limited to these. Said foods, beverages or food additives can be prepared in conventional ways.

[050]The term “health functional food” is a group of foods to which value is added so that the function thereof is exercised and expressed for the predetermined purpose using physical, biochemical or bioengineering techniques thereto or a designed processed food so that the in vivo adjustment functions of the relevant food composition such as rhythm adjustment in prophylaxis, disease prevention and recovery from disease are sufficiently expressed. Such functional food may comprise food supplement additives which are scientifically acceptable in food terms and may further comprise suitable carriers, excipients and diluents, which are commonly used in the manufacture thereof.

[051] The term “functional beverages” as used in the present invention collectively refers to beverage products for quenching thirst or for enjoying taste. There is no particular limitation to these except comprising the composition for improving or preventing the said symptoms of intestinal disease as essential ingredients with the indicated proportion and various flavoring agents or natural carbohydrates may be contained therein as additional ingredients as in ordinary beverages.

[052] In addition to the above, the food comprising the food composition according to the present invention for the amelioration of said symptoms of intestinal disease or the prevention thereof may contain various nutrients, vitamins, minerals (electrolyte), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickening agents, pH adjusting agents, stabilizing agents, preservatives, glycerin, alcohol, carbonizing agents as used in carbonated beverages and the like and each of the above ingredients may be used alone or in combination with each other.

[053] In the food comprising the food composition according to the present invention, the composition of the present invention may be comprised in an amount of 0.001 wt.% to 100 wt.% and preferably 1 wt.% to 99 wt.%, based on the total weight of the food; in the case of beverages, it may be comprised in an amount of 0.001 g to 10 g and preferably 0.01 g to 1 g, based on 100 ml. For long-term intake for health and hygiene purposes or for health control purposes, however, the amount may be below the above-mentioned range; and since the effective ingredients do not have any problems in terms of safety profile, they may be used in an amount above the range and they are not limited to the above-mentioned amount range.

[054] The food composition according to the present invention may comprise strain KBL382, strain KBL384 or strain KBL385 alone or in combination with the acceptable carrier or may be prepared in the form of the composition suitable for consumption by humans or animals. That is, the composition may be added to food that does not contain any probiotic bacteria or some probiotic bacteria. For example, the microorganisms that may be used in combination with the strain according to the present invention in the preparation of the food of the present invention must be suitable for consumption by humans or animals and have probiotic activities to inhibit pathogenic and harmful bacteria or to improve the balance of microorganisms in the mammalian intestinal tract, after entry, but is not limited to these. Such probiotic microorganisms may include, for example, yeasts such as Saccharomyces, Candida, Pichia and Torulopsis, fungi such as Aspergillus, Rhizopus, Mucor and Penicillium and bacteria belonging to the genera of Lactobacillus, Bifidobacterium, Leuconostoc, Lactococcus, Bacillus, Streptococcus, Propionibacterium, Enterococcus and Pediococcus. Specifically suitable probiotic microorganisms may include, for example, Saccharomyces cerevisiae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus delbr uckii, Lactobacillus johnsonii, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus sakei, Lactococcus lactis or Pediococcus acidilactici. Preferably, the food composition according to the present invention may further comprise a mixture of probiotic microorganisms having excellent probiotic activities and superior effects on immunity enhancement to further enhance the effects thereof. Carriers that may be included in the food composition of the present invention may include, for example, extenders, high fiber additives, encapsulating agents and lipids, which are widely known in the art. The Lactobacillus paracasei strain in the present invention may be in lyophilized or encapsulated form or in the form of culture suspensions or dry powders.

[055]The composition of the present invention may also be provided in the form of a feed additive comprising said strain or a feed comprising the same.

[056]The feed additive of the present invention may be in the form of a dry or liquid formulation and may further comprise other non-pathogenic microorganisms in addition to said strain of KBL382, KBL384 or KBL385. Microorganisms that can be added to the feed additive may include, for example, Bacillus subtilis which can produce protease, lipase and sugar converting enzymes, the Lactobacillus strain having physiological activity and degradability of organic compounds under anaerobic conditions such as in the cow's stomach, filamentous fungi such as Aspergillus oryzae which have effects on weight gain of livestock, milk production and feed digestibility (Slyter, L. L. J. Animal Sci,1976, 43, 910 - 926) and yeast such as Saccharomyces cerevisiae (Johnson, D. E et al. J. Anim. Sci.,1983, 56, 735 - 739 ; Williams, P. E. V. et al,1990, 211).

[057]The feed additive of the present invention may additionally comprise at least one enzymatic agent in addition to said Lactobacillus paracasei strain KBL382, Lactobacillus paracasei strain KBL384 or Lactobacillus paracasei strain KBL385. The additional enzymatic agents may be in a dry or liquid form and may include, for example, steatolytic enzymes such as lipase, phytase for producing phosphate and inositol phosphate by degrading phytic acid, amylase, i.e., an enzyme for hydrolyzing the α-1,4-glucoside bond included in, for example, starch and glycogen, phosphatase, i.e., an enzyme for hydrolyzing organic phosphoric acid ester, carboxymethyl cellulase for degrading cellulose, xylase for degrading xylose, maltase for hydrolyzing maltose into two glucose molecules, and sugar-producing enzymes such as invertase for producing the glucose-fructose mixture by hydrolyzing sucrose.

[058] In using Lactobacillus paracasei strain KBL382, Lactobacillus paracasei strain KBL384 or Lactobacillus paracasei strain KBL385 of the present invention as feed additives, raw feed ingredients such as peanuts, peas, beets, pulp, grain by-products, animal viscera powder and fish meal powder, including various grains and soy protein, can be used. They can be processed or not and can be used without limitation. Processing can include, but is not limited to, such a process that the raw feed ingredients are loaded and can be compressed under pressure against a given output and for proteins, extrusion by which proteins are degenerated to increase availability can be preferably used. Extrusion has advantages of denaturing proteins through heat treatment and destroying anti-enzyme factors. In addition, for soy proteins, their digestibility can be improved through extrusion to inactivate antinutrients such as trypsin inhibitor, one of the protease inhibitors present in soybeans. In addition, extrusion can promote the improvement of protease digestibility, enhancing the nutritional value of soy proteins.

[059] In another embodiment, the present invention relates to a pharmaceutical composition for the treatment or prevention of intestinal diseases, comprising an effective amount of at least one selected from the group consisting of microbial cells of a Lactobacillus paracasei strain KBL382, a Lactobacillus paracasei strain KBL384 or a Lactobacillus paracasei strain KBL385, cultures of said strain, lysates of said strain and extracts of said strain.

[060] In another embodiment, the present invention relates to a pharmaceutical composition for the treatment or prevention of allergic diseases, comprising an effective amount of at least one selected from the group consisting of microbial cells of a Lactobacillus paracasei strain KBL382, a Lactobacillus paracasei strain KBL384 or a Lactobacillus paracasei strain KBL385, cultures of said strain, lysates of said strain and extracts of said strain.

[061] In another embodiment, the present invention relates to a pharmaceutical composition for the treatment or prevention of autoimmune diseases or inflammatory diseases, comprising an effective amount of at least one selected from the group consisting of microbial cells of a Lactobacillus paracasei strain KBL382, a Lactobacillus paracasei strain KBL384 or a Lactobacillus paracasei strain KBL385, cultures of said strain, lysates of said strain and extracts of said strain.

[062] The pharmaceutical composition of the present invention may be provided in a form of live bacteria, dried strain, cultures of said strain, lysates of said strain or a composition in combination with a pharmaceutically acceptable carrier or medium. The carriers or media that may be used herein may include solvent, a dispersant, a coating, an enhancer, a controlled release formulation (i.e. sustained release formulation) or at least one inert excipient including starch, polyol, granules, microfine cellulose, microcrystalline cellulose such as Celphere, Celphere pearls, diluent, lubricant, binder, disintegrant and the like. The tablet of the above composition may be, if desired, coated by a standard aqueous or non-aqueous technique. Examples of the pharmaceutically acceptable carrier and excipient for use as the pharmaceutically acceptable inert carrier and said additional ingredients may include, for example, a binder, a filler, a disintegrant, a lubricant, an antimicrobial agent and a coating agent, but are not limited to these.

[063] The present invention can be characterized in that said intestinal diseases are selected from the group consisting of abdominal distension, abdominal discomfort, infectious diarrhea caused by pathogenic microorganisms, gastroenteritis, inflammatory bowel diseases, neurogenic intestinal syndrome, irritable bowel syndrome, overgrowth of microorganisms in the small intestine and intestinal alimentary diarrhea, and the diseases also include those caused by damage to the barrier function of the intestine.

[064]Inflammatory bowel disease (IBD) may include Crohn's disease, the intestinal lesion that accompanies Behcet's disease, ulcerative colitis, bleeding rectal ulcer, and pouchitis and refers to a group of diseases including Crohn's disease and ulcerative colitis. Ulcerative colitis only affects the colon. Inflammation and ulceration from ulcerative colitis are limited to the two innermost layers, mucosa and submucosa of the four layers of the colon. Inflammation and ulceration from Crohn's disease may be diffuse throughout all layers of the intestinal wall in both the small and large intestine.

[065]Meanwhile, irritable bowel syndrome is a chronic condition that accompanies not only persistently recurring abdominal discomfort and pain such as bloating, but also changes in bowel habits such as diarrhea and constipation. Symptoms may be exacerbated by psychological factors or stressful social circumstances.

[066]In the present invention, allergic diseases may include, but are not limited to, atopic dermatitis, urticaria, allergic rhinitis, allergic conjunctivitis, asthma, food allergy, and anaphylactic shock.

[067] In the present invention, autoimmune diseases may include, for example, rheumatoid arthritis, lupus, systemic scleroderma, atopic dermatitis, psoriasis, psoriatic arthritis, asthma, Guillain-Barre syndrome, myasthenia gravis, dermatomyositis, polymyositis, multiple sclerosis, autoimmune encephalomyelitis, polyarteritis nodosa, Hashimoto's thyroiditis, multiple sclerosis, temporal arteritis, juvenile diabetes, alopecia areata, pemphigus, aphthous stomatitis, autoimmune hemolytic anemia, Wegener's granulomatosis, Sjogren's syndrome, Addison's disease, Crohn's disease, and Behcet's disease, but are not limited to these.

[068] Furthermore, inflammatory diseases of the present invention collectively refer to conditions having inflammation as a primary lesion and may include, for example, edema, conjunctivitis, periodontitis, rhinitis, otitis media, chronic sinusitis, pharyngolaryngitis, tonsillitis, bronchitis, pneumonia, gastric ulcer, gastritis, colitis, gout, eczema, acne, atopic dermatitis, contact dermatitis, seborrheic dermatitis, ankylosing spondylitis, rheumatic fever, fibromyalgia, osteoarthritis, psoriatic arthritis, rheumatoid arthritis, periarthritis of the shoulder, tendonitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, Sjogren's syndrome, myasthenia gravis, sepsis, vasculitis, bursitis, temporal arteritis, and multiple sclerosis, but are not limited to these.

[069]It is known that autoimmune diseases can be prevented or treated by activation of regulatory T cells (Treg) or differentiation into T cells. Treg, a type of CD4+ T cell, plays a role in regulating the immune system and controlling autoimmune diseases by maintaining tolerance to self-antigens. A transcription factor called Foxp3 is specifically involved in the development, maintenance and function of regulatory T cells, regulating the immune response through the secretion of the immunosuppressive cytokines IL-10 and TGF-β.

[070]In addition, it is known that when the differentiation rate of T cells into Th1 cells, Th2 cells and Th17 cells is low and the differentiation rate of T cells into Treg cells is high, the inflammatory response can be effectively suppressed.

[071]Th1 cells, Th2 cells and Th17 cells differentiated from splenic T cells are known to be associated with the expression of the following transcription factors and cytokines: In the case of Th1 cells, T-bet, IFN-Y and IL-12; in the case of Th2 cells, GATA3 and IL-5; in the case of Th17 cells, RORYt, IL-17.

[072] The present invention can be characterized in that the effects resulting from the strain of the present invention to alleviate, treat or prevent autoimmune diseases or inflammatory diseases can be induced by at least one selected from the following mechanisms: i) suppression of the expression or secretion of at least one selected from the group consisting of the inflammatory cytokines IL-6, TNF, IL-1b, IL-8, IL-2, IFN-Y, IL-4, IL-13 and IL-17A; ii) increased expression or secretion of the anti-inflammatory and immunoregulatory cytokine IL-10; and iii) increased expression of Treg involved in anti-inflammatory activities and immunoregulation.

[073]The term ‘treating’, unless otherwise stated, refers to reversing or alleviating the diseases or conditions used with said term or one or more symptoms thereof, inhibiting the progression thereof or preventing the same. The term ‘treatment’ as used in the present invention refers to an action of ‘treating’ as defined above. Accordingly, treatment or therapeutic regimen for intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases in mammals may include one or more of the following: (1) Inhibiting the development of intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases, (2) Preventing the spread of intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases, (3) Alleviating intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases, (4) Preventing the recurrence of intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases, and (5) Alleviating the symptoms of intestinal diseases, allergic diseases, autoimmune diseases, or inflammatory diseases.

[074] A composition of the present invention for preventing or treating intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases may comprise a pharmaceutically effective amount of a Lactobacillus paracasei strain KBL382, a Lactobacillus paracasei strain KBL384, a Lactobacillus paracasei strain KBL385 alone or in combination with at least one of pharmaceutically acceptable carriers, excipients or diluents.

[075] In the present invention, the term “effective amount” means an amount that is high enough to provide a desired effect but is low enough to prevent serious side effects under medical judgment. The amount of microorganisms administered to the body by the composition of the present invention can be appropriately adjusted in consideration of the route of administration and the target of administration.

[076] The composition of the present invention can be administered to a subject once or more per day. Unit dosage means physically separate units suitable for unitary administration to human subjects and other mammals and each unit comprises a suitable pharmaceutical carrier and a predetermined amount of the microorganisms of the present invention to provide a therapeutic effect. The dosage unit for oral administration to an adult patient preferably contains 0.001 g or more of the microorganism of the present invention and the oral dosage of the composition of the present invention is 0.001 g to 10 g and preferably 0.01 g to 5 g per dose. The pharmaceutically effective amount of the microorganism of the present invention is 0.01 g to 10 g/day. However, the dosage varies depending on the severity of the patient's disease and the microorganisms and effective auxiliary ingredients used together. Furthermore, the total daily dosage can be divided into several times and continuously administered as needed. Accordingly, the above dosage ranges do not limit the scope of the present invention in any way.

[077]Furthermore, the term “pharmaceutically acceptable” as used above refers to a composition that is physiologically acceptable and does not cause an allergic reaction such as gastrointestinal upset, dizziness or similar reaction when administered to a human being.

[078] A composition of the present invention may be formulated using methods known in the art so that rapid, sustained or delayed release of the active ingredients after administration to a mammal may be provided. Dosage forms may be powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injection solutions or sterile powders. Furthermore, the composition of the present invention for preventing or treating intestinal diseases may be administered via various routes, including oral, transdermal, subcutaneous, intravenous or intramuscular administration. The dosage of the active ingredients may be appropriately selected depending on various factors such as the route of administration, the patient's age, sex, weight and the severity of the patient's condition. The composition of the present invention for preventing or treating gastrointestinal diseases may be administered in combination with a known compound having the effect of preventing or treating the symptoms of intestinal diseases.

[079] The pharmaceutical composition of the present invention, in particular, may be provided in an oral unit dosage form of an enteric coated formulation. The term “enteric coating” as used herein comprises any known pharmaceutically acceptable coating that can remain in the stomach without being degraded by gastric acid and can sufficiently disintegrate in the intestinal tract to release the active ingredients therein. The “enteric coating” of the present invention refers to a coating that can be maintained for 2 hours or more when an artificial gastric juice such as a pH 1 HCl solution is contacted therewith at 36°C to 38°C and can subsequently degrade, preferably in an artificial intestinal juice such as a pH 6.8 KH2PO4 buffer solution within 30 minutes.

[080] The enteric coating of the present invention is coated on a core in an amount of about 16 mg to 30 mg, preferably 16 mg to 20 mg or 25 mg or less. When the thickness of the enteric coating of the present invention is 5 μm to 100 μm and preferably 20 μm to 80 μm, satisfactory results can be obtained as an enteric coating. The enteric coating material can be suitably selected from known polymeric materials. Suitable polymeric materials are listed in various known documents [L. Lachman et al., The Theory and Practice of Industrial Pharmacy, 3rd ed., 1986, pp. 365 ~ 373; H. Sucker et al., Pharmazeutische Technologie, Thieme, 1991, pp. 355 - 359; Hagers Handbuchder pharmazeutischen Praxis, 4th ed., Vol. 7, pages 739~742 and 766~778, (SpringerVerlag, 1971); and Remington’s Pharmaceutical Sciences, 13th ed., pages 1689~1691 (Mack Publ., Co., 1970)] and cellulose ester derivatives, cellulose ethers, a copolymer of acrylic resin and methyl acrylate, and a copolymer of maleic acid and phthalic acid derivatives may be included herein.

[081] The enteric coating of the present invention can be prepared using a conventional enteric coating method in which an enteric coating solution is sprayed onto a core. Suitable solvents used for the enteric coating process are alcohols such as ethanol, ketones such as acetone, halogenated hydrocarbon solvents such as dichloromethane (CH2Cl2) and mixed solvents of these solvents. A softener such as di-n-butyl phthalate or triacetin is added to the coating solution in a ratio of 1:about 0.05 to about 0.3 (coating material:softener). It is suitable to carry out the spraying process continuously and it is possible to adjust the spraying amount in consideration of the coating conditions. The spraying pressure can be variably adjusted and satisfactory results can generally be obtained with a spraying pressure of about 1 bar to about 1.5 bar.

[082] In another embodiment, the present invention relates to the use of said strain or composition for the prevention or treatment of intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases and to the use of said strain or composition to prepare a therapeutic agent for intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases.

[083] Specifically, the present invention relates to a composition for the use of preventing or treating intestinal diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[084] Furthermore, the present invention relates to a composition for the use of preventing or treating allergic diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[085] Furthermore, the present invention relates to a composition for the use of preventing or treating autoimmune diseases or inflammatory diseases, comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain.

[086] The present invention also relates to the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for intestinal diseases.

[087] Furthermore, the present invention relates to the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for allergic diseases.

[088] Furthermore, the present invention relates to the use of a composition comprising at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain, to prepare a preventive or therapeutic drug for autoimmune diseases or inflammatory diseases.

[089]The term ‘prevention’, as used here, is associated with preventing, delaying, impeding or inhibiting diseases in order to reduce them.

[090]The term ‘treatment’, as used here, is associated with the care of an individual suffering from diseases in order to improve, cure or reduce the symptoms of the diseases or reduce or stop the progression of the diseases.

[091]In another embodiment, the present invention provides a method for preventing or treating intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases, comprising administering a pharmaceutically effective amount of said strain or said composition to a subject in need of prevention or treatment of said diseases or in need of alleviation of intestinal health, allergic reactions, decreased level of immunity or inflammatory reactions.

[092] Specifically, the present invention provides a method for treating intestinal diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511 BP), cultures of said strain, lysates of said strain and extracts of said strain to a subject in need thereof.

[093] Further, the present invention provides a method for treating allergic diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain to a subject in need thereof.

[094] Further, the present invention provides a method for treating autoimmune diseases or inflammatory diseases, comprising administering a therapeutically effective amount of at least one selected from the group consisting of a Lactobacillus paracasei strain KBL382 (Accession No. KCTC13509BP), a Lactobacillus paracasei strain KBL384 (Accession No. KCTC13510BP) or a Lactobacillus paracasei strain KBL385 (Accession No. KCTC13511BP), cultures of said strain, lysates of said strain and extracts of said strain to a subject in need thereof.

[095] Since the pharmaceutical composition used for the method for preventing or treating said intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases and the method of administration thereof have been described above, the overlapping levels between the composition and the method will be omitted here to avoid excessive complexity of the present descriptive report.

[096]Meanwhile, said individual to whom the composition for preventing or treating said intestinal diseases, allergic diseases, autoimmune diseases or inflammatory diseases may be administered includes all animals including humans. For example, the individual may be an animal such as a dog, cat or mouse.

[097]The present invention will now be described in more detail by means of examples. These examples are intended to illustrate the present invention only and it will be apparent to those skilled in the art that the scope of the present invention should not be construed as being limited by these examples.

Example 1. Screening of probiotic strains with immunoregulatory function

[098] Probiotic strains having an immunoregulatory function were screened using the THP-1 monocytic leukemia cell line and peripheral blood mononuclear cells (PBMC). Strains derived from the human intestine or vagina, respectively, were seeded to the above two cell lines so that the cell line:strain ratio was 1:100, and the ratio of IL-10, a major cytokine indicating inflammation control, to IL-6, a major cytokine indicating an inflammatory reaction (IL-10/IL-6), and the ratio of IL-10 to IFN-Y, a major cytokine indicating autoimmune reaction (IL-10/IFN-Y) were measured. A total of 23 strains were used for screening as follows: two strains of Lactobacillus gasseri, one strain of Lactobacillus reuteri, five strains of Lactobacillus rhamnosus, two strains of Lactobacillus fermentuma, four strains of Lactobacillus paracasei, four strains of Lactobacillus salivarius, one strain of Lactobacilus plantarum, two strains of Lactobacillus acidophilus, and two strains of Lactococcus lactis.

[099] As a result of the screening, the highest IL-10/IL-6 values were shown in the three Lactobacillus paracasei strains KBL382, KBL384 and KBL385. In particular, among the three Lactobacillus paracasei strains, KBL382 had the highest IL-10/IL-6 value of 2.22 in THP-1 cell line and the IL-10/IFN-Y value was 9 in peripheral blood mononuclear cells, showing a very high immunoregulatory effect. Table 1 below shows the results of the IL-10/IL-6 value and IL-10/IFN-Y value measured for three Lactobacillus paracasei strains out of the twenty-three probiotic strains used in the present screening. Consequently, three strains of Lactobacillus paracasei KBL382, KBL384 and KBL385 were predicted to have a high effect on improving intestinal health and further experiments were conducted on these strains.

Example 2. Verification of the effect of Lactobacillus paracasei strains KBL382, KBL384 and KBL385 on inflammatory cytokine expression and regulation of T cell differentiation (in vitro test)

[0100] In order to verify the immunoregulatory effect of said Lactobacillus paracasei strains KBL382, KBL384 and KBL385, the ability to generate cytokines involved in immunoregulation was confirmed in the THP-1 cell line treated with Lactobacillus paracasei strains KBL382, KBL384 and KBL385, while whether the T cell differentiation marker gene was expressed was confirmed in PBMC treated with Lactobacillus paracasei strains KBL382 and KBL385.

[0101]First, the THP-1 cell line was seeded onto each well of a 24-well plate by 1 x 105 cells, differentiated into mature macrophages, and then the culture solution was replaced. After 3 hours, one well was treated with LPS at a concentration of 1 μg/mL as a positive control, and the other wells were treated with Lactobacillus paracasei strain KBL382, KBL384, or KBL385, each by 1 x 107 cells based on the number of viable bacteria; after 24 hours, the culture solution was collected, and the amount of each cytokine was measured using the BD Cytometric Bead Array (CBA) Human Inflammation Kit (Cat No. 551811) according to the manufacturer's method. As a negative control (Cont), the well was treated with PBS buffer, and the same experimental procedure was performed. As a result, as shown in FIG. 1, the measurement showed that a group of THP-1 cell lines treated with Lactobacillus paracasei strains KBL382 and KBL385 had a significantly lower amount of inflammatory cytokines IL-6, TNF, IL-1b, and IL-8 than the LPS-treated group. In the case of treatment with strain KBL384, it was confirmed that IL-6 was significantly lower than the LPS-treated test group, but TNF, IL-1b, and IL-8 were not significantly different from the LPS-treated group. In the case of IL-10, no significant increase or decrease was observed in the KBL382, KBL384, and KBL385-treated groups compared to the LPS-treated group, but it was observed that the overall increase in the IL-10 level was induced compared to the negative control group. Therefore, it was concluded that Lactobacillus paracasei strains KBL382 and KBL385 were effective in significantly suppressing the generation of inflammatory cytokines in THP-1 cells and the subsequent experiment was conducted to verify the T cell differentiation pattern.

[0102] In order to confirm the effect of KBL382 and KBL385 strains on T cell differentiation, PBMC cells were seeded onto each well of a 24-well plate by 5 x 105 cells and then Lactobacillus paracasei strain KBL382 or KBL385 was added by 5 x 106 cells. As a control, E. coli (E. coli 0157 EC4115) was added to each well by 5 x 106 cells based on the number of viable cells or LPS was added at a concentration of 500 ng/mL. E. coli is known to increase the expression of T-bet, GATA3 and RORYt genes, which are T cell effector cell markers related to the inflammatory response, while LPS is known to increase the expression of FOXP3 gene, a marker of Treg cells related to the regulation of inflammation. The PBMC cell test group prepared under the above conditions was incubated for 5 days, and then the cells were obtained and collected. In order to confirm the gene expression level, first, the easy-spinTM Total RNA Extraction Kit (Intron) (DNA-free) was used to extract RNA, and then the High Capacity RNA-to-cDNA Kit (ThermoFisher) was used to synthesize cDNA. A real-time PCR was conducted on the cDNA synthesized with the Rotor-Gene SYBR Green PCR Kit (Qiagen) using a Rotor-Gene® Q instrument (Qiagen) according to the manufacturer's method, and the mRNA expression of T-bet which is the Th1 marker gene (effector cell marker), GATA3 which is the Th2 marker gene, RORYt which is the Th17 marker gene, and FOXP3 which is the Treg cell marker gene were measured. At this time, the expression level of B2M genes was measured as an internal control to correct the relative gene expression level between each test group. The base sequence of the primer used to confirm the expression of each gene is as follows. B2M Forward: 5’-CCA GCA GAG AAT GGA AAG TC-3’’(SEQ ID NO. 4) Reverse: 5’-GAT GCT TCT TAC ATG TCT CG-3’ (SEQ ID NO. 5) T-bet Forward: 5’-CCC CAA GGA ATT GAC AGT TG-3’ (SEQ ID NO. 6) Reverse: 5’-GGG AAA CTA AAG CTC ACA AAC-3' (SEQ ID NO. 7) GATA3 Forward: 5'-CTG CAA TGC CTG TGG GCT C-3' (SEQ ID NO. 8) Reverse: 5'-GAC TGC AGG GAC TCT CGC T-3' (SEQ ID NO. 9) RORYt Forward: 5'-AAG AGE CAT CGC CAA AGC AT-3' (SEQ ID NO. 10) Reverse: 5'-TCC ACA TGC TGG CTA CAC UM-3' (SEQ ID NO. 11) FOXP3 Forward: 5'-TCA AGC AGE GCC AGG CG-3' (SEQ ID NO. 12) Reverse: 5'-CAG GAG CCC TTG TCG GAT-3' (SEQ ID NO. 13)

[0103] As a result, as shown in FIG. 2, it was confirmed that Lactobacillus paracasei strains KBL382 and KBL385, compared to E. coli, maintained the expression level of T-bet, GATA3, and RORYt genes, which are effector cell markers of inflammatory T cells, at a significantly low level, but significantly increased mRNA of FOXP3, a marker of Treg cells to maintain immune homeostasis similarly to LPS.

Example 3. Verification of the effect of Lactobacillus paracasei strains KBL382, KBL384 and KBL385 on T cell immunoregulation (in vitro test)

[0104]Meanwhile, in order to verify the immunoregulatory effect of T cell by Lactobacillus paracasei strains KBL382, KBL384 and KBL385, the expression level of inflammatory cytokines was checked when T cells were activated in PBMC cells and then followed by treatment with said Lactobacillus paracasei strains KBL382, KBL384 and KBL385.

[0105]PBMC cells were seeded onto each well of a 96-well plate by 2 x 105 cells with an anti-CD3 antibody (1 μg/mL, ebioscience) to activate T cells and then a Lactobacillus paracasei strain KBL382, KBL384 or KBL385 was added by 1 x 107 cells to be “bacteria:PBMC = 50:1”, based on the number of viable cells. As a control, Escherichia coli K12 (ATCC 10798) was added to each well by 1 x 107 cells based on the number of viable cells. After incubating the prepared T cell-activated PBMC cell test group under the above conditions for 3 days, the supernatant was collected and the amount of each cytokine was measured using the BD Cytometric Bead Array (CBA) Human Inflammation Kit (Cat No. 551811) according to the manufacturer's method.

[0106] As a result, as shown in FIG. 3 , in the PBMC cell lines so that T cells were activated by adding an anti-CD3 antibody, all Lactobacillus paracasei strains KBL382, KBL384, and KBL385 significantly decreased the expression of IL-2, which is a Th1 cytokine, IL-4 and IL-13, which are respectively IFNY and Th2 cytokines, and IL-17A, which is a Th17 cytokine, compared to the test group treated with only the E. coli-added strain and PBS, while the anti-inflammatory cytokine IL-10 was markedly increased in all three Lactobacillus paracasei strains used in the experiment, compared to the test group treated with only PBS; it was confirmed that especially strain KBL 382, among the three, significantly increased IL-10 compared to the E. coli-added group. The experiment results as above led to a conclusion that all Lactobacillus paracasei strains KBL382, KBL384 and KBL385 significantly decreased T-cell inflammatory cytokines and increased anti-inflammatory cytokines.

Example 4. Verification of the effect of Lactobacillus paracasei strains KBL382, KBL384 and KBL385 on enteritis alleviating effects (in vivo test)

[0107]The present example was to confirm whether Lactobacillus paracasei strains KBL382, KBL384 and KBL385 showed the effects of improving intestinal function even in vivo. For this purpose, C57BL/6 mice were divided into groups of 10 mice each and then fed with tap water with 2% DSS dissolved therein for 9 days and thereby inducing enteritis. At the same time, mice in the control group were orally administered with 200 μL of PBS daily and mice in the test group were daily provided via oral administration with 200 μL each of Lactobacillus paracasei strains KBL382, KBL384 or KBL385 which was diluted in PBS to be 2 x 1010 CFU/mL so that the daily administration amount could be adjusted to 4 x 109 CFU. Then, during the 9 days that enteritis was induced by DSS, the body weight changes of the mice in the control group and test group were measured daily, and on the 9th day after DSS was given, the mice were subjected to autopsies to measure the colon length.

[0108]As a result, as shown in FIG. 4, it was confirmed that with respect to the changes of body weight, the groups treated with strains KBL382, KBL384 and KBL385 all showed a significant effect on alleviating weight loss, compared with the mice in the control group without any treatment; with respect to the change in colon length, the amplitude of the decrease in colon length was significantly improved in all groups of three strains, compared with the mice in the control group.

[0109]Furthermore, H&E staining was performed to measure the degree of inflammation in the test groups of KBL382 and KBL385 strains and Myeloperoxidase (MPO) levels were measured in the tissues. After autopsy, in order to examine the colon lesions by H&E staining, the distal part of the colon was fixed in 10% neutral formalin solution and then paraffin tissue specimens were cut at a thickness of 5 μm, stained with hematoxylin & eosin and observed with a light microscope. Furthermore, for the measurement of MPO, the colon tissue was first placed in RIPA buffer to which a protease inhibitor was added and was disrupted with a homogenizer. After centrifuging the disrupted tissue at 4 °C. at 15,000 x g for 10 min, MPO was measured using the supernatant according to the ELISA kit protocol (Hycult Biotech, MPO, Mouse, cat. HK210-02).

[0110] As a result, as shown in FIG. 4, it was confirmed through the H&E staining results that all the test groups that the two strains, KBL382 and KBL385, were administered, had alleviated inflammation compared to the control group, and the level of MPO was slightly decreased in the KBL385 test group and significantly decreased in the KBL382 test group. Using the colon tissues thus obtained were used to confirm the strengthening effect on the tight junction of the intestinal tract wall.

Example 5. Effects of Lactobacillus paracasei strains KBL382 and KBL385 on strengthening the tight junction of the intestinal tract wall

[0111] In order to verify the effects of Lactobacillus paracasei strains KBL382 and KBL385 on the strengthening of the tight junction of the intestinal tract wall, the mRNA expression levels of genes involved in the tight junction of the intestinal tract wall were compared and measured in the cells isolated from the mouse colon tissue isolated in Example 4. To verify the amount of gene expression, RNA was first extracted from the tissues using the easy-spinTM Total RNA Extraction Kit (Intron) (DNA-free) and then cDNA was synthesized with the High Capacity RNA-to-cDNA Kit (ThermoFisher). The Rotor-Gene SYBR Green PCR kit (Qiagen) was used to measure in vivo the mRNA expression level of zonula occluden-1 (ZO-1), Claudin3 and MUC-4, which are the marker proteins of the tight junction of the intestinal tract wall, using a Rotor-Gene® Q (Qiagen). The expression level of the hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene was measured as a control to normalize the relative gene expression level between each test group. The primer used to confirm the expression is prepared to be able to specifically bind to each gene as follows. Zo-1 Fw: 5'-ACC CGA AAC TGA TGC TGT GGA TAG-3' (SEQ ID NO. 14) GC TGG AGE TGA AC-3’ (SEQ ID NO. 17) MUC-4 Fw: 5’-GTC TCC CAT CAC GGT TCA GT-3’ (SEQ ID NO. 18) Rw: 5’-TGT CAT TCC ACA CTC CCA GA-3’ (SEQ ID NO. 19) HPRT Fw: 5’-TTA TGG ACA GGA CTG AAA GAC-3’’ (SEQ ID NO. 20) Rw: 5’-GCT TTA ATG TAA TCC AGC AGG T-3’’ (SEQ ID NO. 21)

[0112] As a result, as shown in FIG. 5, it was found that the levels of the three genes measured in the mice dosed with the KBL382 and KBL385 strains were increased compared to the control group dosed with PBS alone and thus the tight junction of the intestinal tract wall was recovered; consequently, said strains can be used to treat diseases such as IBD and IBS.

Example 6. Comparison of the enteritis-alleviating effects of an enteritis therapeutic antibody and KBL382

[0113]Infliximab (product name, Remicade) is a recombinant antibody therapeutic drug used as an injection for autoimmune diseases such as rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, pediatric Crohn's disease, psoriasis, and psoriatic arthritis. The purpose of this study was to confirm the difference in the effect on alleviating symptoms of intestinal disease in vivo between the KBL 382 strain and infliximab.

[0114]For comparison of the effects, after dividing the C57BL/6 mice into groups of eight mice, the mice were fed with tap water with 2% DSS dissolved therein for 9 days, inducing enteritis. At the same time, 200 μL of PBS was orally administered to the mice in the control group daily, while 200 μL of the KBL382 strain diluted in PBS to be 2 x 1010 CFU/mL was orally administered to the mice in the test group daily so that the daily administration amount could be adjusted to 4 x 109 CFU. To the therapeutic antibody administration group, the infliximab antibody was administered once on Day 3 to be a dose of 5 mg/kg per mouse.

[0115]Later, during the 9 days that enteritis was induced by DSS, the body weight changes of the mice in the control group and test group were measured daily and on the 9th day after DSS was given, the mice were subjected to autopsies to measure the colon length.

[0116]As a result, as shown in Fig. 6(A), with respect to the change in colon length, it was confirmed that the amplitude of the decrease in colon length was significantly improved in the groups that the KBL382 strain was treated and that infliximab was administered, compared with the mice in the control group. It was confirmed that the effect of improving the colon length of the group that the KBL382 strain was treated was similar to or slightly higher than that of the group that infliximab was administered.

[0117] With respect to the change in body weights, as shown in Fig. 6(B), the effect on decreasing body weight was significantly improved in the groups that the KBL382 strain was treated and that infliximab was administered, compared to the mice in the control group without any treatment. It was confirmed that the effect of alleviating initial weight loss for five days after administration was superior in the infliximab administered group over the KBL382 strain treated group, but later, the ameliorating effect by infliximab was decreased, while the effect of the KBL382 strain treated group on weight loss was increased, showing similar effects between the two groups until the 9th day. Consequently, with respect to the use of the said strain for the treatment of inflammatory bowel diseases and irritable bowel syndrome, it was confirmed that a therapeutic effect similar to commercially available therapeutic antibodies can be expected.

Example 7. Effects of KBL382 on the relief of atopic conditions

[0118]In order to verify the effect on atopic alleviation of the KBL382 strain, the NC/Nga mouse model, an animal model of atopic skin disease, was used.

[0119]After dividing the NC/Nga mice into groups of nine mice, the back of each mouse was shaved from the lower ear to the upper tail, and the mice were left for 24 hours. Then, an ointment comprising Dermatophagoides farinae extract (DFE), a house dust mite (HDM) antigen, was applied once or twice a week for seven weeks on the shaved portion so that each mouse could be treated with 100 mg of DFE and thereby inducing atopic dermatitis. From the third week of dermatitis induction, 200 μL of PBS was orally administered to the mice in the control group daily; each of Lactobacillus rhamnosus strain KBL382 and strain KBL365, which were diluted in 200 μL of PBS to be at least 1 × 109 CFU/mouse, were orally administered to the mice in the test group by 200 μL daily. Then, during four weeks of administration of the bacteria, the dermatitis scores of the mice in the control group and test group were measured weekly, and at the 4th week after administration of the bacteria, the scratching time and skin thickness of the mice and the blood IgE concentration after conducting autopsies of the mice were measured. 7-1. Evaluation of dermatitis score

[0120]To assess DFE-induced skin lesions, the dermatitis score was measured by the following method. Skin conditions were monitored by taking images for 4 weeks at one-week intervals starting from the 3rd week since the strain was administered. Four indicators of dryness, edema, erythema/hemorrhage and erosion/excoriation of the skin were checked. The total score was evaluated with 0 points for no lesion, 1 point for mild, 2 points for moderate and 3 points for severe.

[0121]As a result, as shown in FIG. 7(A), the dermatitis score was significantly reduced in the group dosed with the KBL382 strain, compared with the control (negative) group where atopic dermatitis was induced and the group that the KBL365 strain was administered. As a result, the effect of alleviating the atopic dermatitis score according to the input of the KBL382 strain was verified. 7-2 Effects on Alleviating Itching Symptoms

[0122] In order to verify the itch-relieving effect of the administration of KBL382 strain in mouse models suffering from DFE-induced atopic dermatitis, the number of scratches was measured for the mouse during 10 minutes after 4 weeks of the administration of the strain.

[0123]As a result, as can be seen in FIG. 7(B), it appeared that the number of scratches was significantly reduced in the test group administered KBL382, compared to the group administered PBS, which confirmed that the itching symptoms of atopic dermatitis were alleviated by the administration of the KBL382 strain. 7-3. Decrease in Skin Thickness

[0124] In order to verify the effect of decreasing skin thickness after administration of KBL382 to mouse models suffering from DFE-induced atopic dermatitis, the mouse ear thickness and dorsal skin thickness were measured with calipers four weeks after the strain was administered and the relief of the edema symptom due to atopic dermatitis was observed. As a result, as can be seen in FIG. 7(C), it was observed that in the test group dosed with KBL382, the ear and skin thicknesses were significantly reduced, compared to the control group and the group administered with KBL365. 7-4. Decrease in Blood IgE Concentration

[0125] It was found that the IgE concentration in patients having atopic dermatitis increased mainly as the clinical severity of atopic dermatitis increased (Matsumoto M, J. Immunol. 1999). Therefore, the concentration of IgE, a representative hematological factor appearing as atopic dermatitis arises, was measured by collecting blood three weeks after the strain was administered, separating the serum therefrom, and using the Mouse IgE ELISA Set (Cat No. 555248, BD OptEIA™). As a result, as shown in FIG. 7(D), it was found that the blood IgE concentration was significantly decreased in the test group that KBL382 was orally administered, which indicated the treatment effect of atopic dermatitis after the administration of KBL382.

[0126] The specific aspects of the present invention have been described in detail above and it is obvious to those skilled in the art that these specific aspects are only preferred embodiments and the scope of the present invention is not limited thereby. Therefore, the scope of the present invention is substantially defined by the following claims, with equivalents to the claims. Name of the Depositary Organization: Korea Research Institute of Bioscience & Biotechnology Accession No.: KCTC13509BP Accession Date: 20180417 Name of the Depositary Organization: Korea Research Institute of Bioscience & Biotechnology Accession No.: KCTC13510BP Accession Date: 20180417 Name of the Depositary Organization: Korea Research Institute of Bioscience & Biotechnology Accession No.: KCTC13511BP Accession Date: 20180417

Industrial Applicability

[0127] The strains of Lactobacillus paracasei KBL382 (Accession No. KCTC13509BP), Lactobacillus paracasei KBL384 (Accession No. KCTC13510BP) and Lactobacillus paracasei KBL385 (Accession No. KCTC13511BP) according to the present invention have excellent anti-inflammatory and immunomodulatory functions, have an excellent strengthening effect on the tight junction of the intestinal tract wall, suppress enteritis-induced weight loss and shorten the length of the colon, thereby exhibiting a therapeutic effect for enteritis and weakening the allergic reaction of cells and significantly alleviating the symptoms of atopic dermatitis and thereby providing improvement of allergic diseases. Therefore, a single strain can achieve enhancement of anti-inflammatory effects, strengthening of immunity, improvement of intestinal health functions and alleviation of allergic diseases, and thus may be useful as a probiotic material.

Sequence Listing Free Text

[0128]An electronic file of the attached Sequence Listing

Claims (19)

1. Strain CHARACTERIZED by the fact that it is Lactobacillus paracasei KBL382 with Accession No. KCTC13509BP. 2. The strain of claim 1, wherein said strain comprises the 16s rDNA sequence of SEQ ID NO: 1. 3. Strain CHARACTERIZED by the fact that it is Lactobacillus paracasei KBL384 with Accession No. KCTC13510BP. 4. The strain of claim 3, wherein said strain comprises the 16s rDNA sequence of SEQ ID NO: 2. 5. Strain CHARACTERIZED by the fact that it is Lactobacillus paracasei KBL385 with Accession No. KCTC13511BP. 6. The strain of claim 5, wherein said strain comprises the 16s rDNA sequence of SEQ ID NO: 3. 7. Food composition CHARACTERIZED by the fact that it comprises an effective amount of at least one selected from the group consisting of the strain, as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain. 8. Food additive composition CHARACTERIZED by the fact that it comprises an effective amount of at least one selected from the group consisting of the strain, as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain. 9. Food composition, according to claim 7, CHARACTERIZED by the fact that said composition is a healthy functional food composition for improving intestinal health. 10. Pharmaceutical composition for the treatment or prevention of intestinal diseases, CHARACTERIZED by the fact that it comprises an effective amount of at least one selected from the group consisting of the strain, as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain. 11. Pharmaceutical composition, according to claim 10, CHARACTERIZED by the fact that said intestinal diseases are selected from the group consisting of abdominal distension, abdominal discomfort, infectious diarrhea caused by pathogenic microorganisms, gastroenteritis, inflammatory bowel diseases, neurogenic bowel syndrome, irritable bowel syndrome, overgrowth of microorganisms in the small intestine and alimentary intestinal diarrhea. 12. Food composition, according to claim 7, CHARACTERIZED by the fact that said composition is a healthy functional food composition to improve allergic symptoms. 13. Pharmaceutical composition for the treatment or prevention of allergic diseases, CHARACTERIZED by the fact that it comprises an effective amount of at least one selected from the group consisting of the strain, as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain. 14. Pharmaceutical composition, according to claim 13, CHARACTERIZED by the fact that said allergic disease is atopic dermatitis, urticaria, allergic rhinitis, allergic conjunctivitis, asthma, food allergy or anaphylactic shock. 15. Food composition, according to claim 7, CHARACTERIZED by the fact that said composition is a healthy functional food composition to alleviate autoimmune diseases or inflammatory diseases. 16. Pharmaceutical composition for the treatment or prevention of autoimmune diseases or inflammatory diseases, CHARACTERIZED by the fact that it comprises an effective amount of at least one selected from the group consisting of the strain, as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain. 17. Use of a composition comprising at least one selected from the group consisting of the strain as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain, CHARACTERIZED by the fact that it is for preparing a drug for preventing or treating intestinal diseases. 18. Use of a composition comprising at least one selected from the group consisting of the strain as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain, CHARACTERIZED by the fact that it is for preparing a drug for preventing or treating allergic diseases. 19. Use of a composition comprising at least one selected from the group consisting of the strain as defined in any one of claims 1 to 6, cultures of said strain, lysates of said strain and extracts of said strain, CHARACTERIZED by the fact that it is for preparing a drug for preventing or treating autoimmune diseases or inflammatory diseases.