Bifidobacterium longum subsp. infantis-derived microvesicles and uses thereof

Abstract

The finding that microvesicles derived from B. infantis, which is an anaerobic intestinal bacterium, have a prophylactic and therapeutic effect against inflammatory diseases has led to the present invention. According to the present invention, the active ingredient can be effectively delivered to intestinal epithelial cells, which are therapeutic targets for inflammatory bowel disease, thereby providing excellent therapeutic effects. In addition, the intestinal lactic acid bacteria-derived microvesicles of the present invention are derived from bacteria in the body, and thus exhibit excellent stability and are less likely to cause side effects when administered for a long period of time.

Description

Microvesicles derived from BIFIDOBACTERIUM LONGUM SUBSP. INFANTIS and their uses

[0001] The present invention relates to the use of microvesicles derived from intestinal bacteria for the treatment of inflammatory diseases.

[0002] Inflammatory bowel disease (IBD) is an intractable disease characterized by chronic inflammation of the gastrointestinal tract with repeated flare-ups and remissions of unknown cause. Crohn's disease (CD) and ulcerative colitis (UC) are known as representative diseases. Although it is a disease mainly common in Caucasians in North America and Northern Europe, the incidence of IBD has been increasing in Korea recently due to factors such as a westernized diet, improved hygiene, and a decrease in infectious diseases. Unlike other chronic diseases, IBD has a high risk of developing in young people, and its cause is unknown, making treatment difficult. In moderate to severe cases, steroids administered can cause serious side effects (elevated blood pressure / blood sugar, acne, weight gain, increased facial hair, cataracts, etc.). For patients dependent on steroids, immunomodulators of the thioprine series can be administered, but this can cause serious complications such as bone marrow suppression, tumor development, and adverse drug reactions in the liver and kidneys. In cases where symptoms are not controlled even with step-by-step treatment with aminosalicylic acid, steroids, and immunomodulators, biological agents can be administered, but problems such as opportunistic infections, hypersensitivity reactions, and decreased effectiveness due to immune function with long-term use can occur, making treatment difficult.

[0003] Therefore, in order to treat patients with inflammatory bowel disease who require long-term medication, a treatment strategy that prevents disease exacerbation and recurrence through intestinal mucosal healing and improves stability is required.

[0004] Extracellular vesicles (EVs) are nano-sized (50-150 nm) vesicles secreted from cells for the purpose of information transfer. These EVs contain various types of genetic material, proteins, and peptides for information transfer, and have the characteristic of easily transmitting information into the cell by binding to the cell membrane. Bacterial nanovesicles are separated from the bacterial cell membrane and are known to contain not only bacterial cell membrane components, but also genetic material such as bacterial DNA and RNA, membrane proteins, and pathogenic proteins. Bacterial nanovesicles function as information carriers such as the transfer of proteins or genetic material between homologous species, cell signaling, and contribute to the elimination of competing organisms or enhancing bacterial survival. In addition, they have the potential to modulate the pathogenesis of bacterial diseases by delivering toxins to the host.

[0005] The technical problem to be achieved by the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory diseases, which comprises microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0006] Another technical problem to be achieved by the present invention is to provide a food composition for preventing or improving inflammatory diseases, which comprises microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0007] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0008] To solve the above problem, the present inventors provide a pharmaceutical composition for preventing or treating inflammatory diseases, which comprises microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0009] According to one side, the microvesicles may have an average diameter of 50 to 150 nm.

[0010] According to one side, the microvesicles may be naturally or artificially secreted from Bifidobacterium longumsubsp.infantis.

[0011] According to one aspect, the microvesicles may be isolated from a culture solution of Bifidobacterium longumsubsp.infantis or from food cultured with the addition of Bifidobacterium longumsubsp.infantis.

[0012] According to one aspect, the inflammatory disease may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease, and inflammatory bowel disease.

[0013] According to another embodiment of the present invention, a food composition for preventing or improving inflammatory bowel disease is provided, comprising microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0014] According to another embodiment of the present invention, a method for producing microvesicles derived from Bifidobacterium longum subsp. infantis is provided, comprising: 1) culturing Bifidobacterium longum subsp. infantis under anaerobic conditions to obtain a culture solution; 2) centrifuging the culture solution; and 3) separating microvesicles through tangential flow filtration (TFF).

[0015] According to another specific example of the present invention, microvesicles derived from Bifidobacterium longumSubspeciesinfantis, manufactured by the above manufacturing method, are provided.

[0016] According to the present invention, bacterial-derived nanovesicles have a much higher ability to induce a host immune response compared to a single antigen. Furthermore, their small size and ability to deliver multiple toxins simultaneously facilitate their delivery to host cells, resulting in excellent therapeutic efficacy. Furthermore, given the nature of inflammatory bowel disease, long-term drug administration and treatment are necessary. The intestinal beneficial bacteria-derived microvesicles of the present invention, derived from intestinal bacteria, exhibit excellent stability and have minimal risk of adverse effects during long-term administration.

[0017] The effects of the present invention are not limited to those mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

[0018] Figure 1 shows the anaerobic bacteria cultivation process in the present invention.

[0019] Figure 2 shows the results of analyzing the characteristics of nanovesicles derived from intestinal beneficial bacteria.

[0020] Figure 3 shows the expression level of inflammatory cytokine mRNA after treating colon cancer epithelial cell lines with B. infantis-EV for 24 hours.

[0021] Figure 4 shows the results of analyzing phenotype and inflammatory cytokine expression to confirm the anti-inflammatory effect through regulation of macrophage polarization.

[0022] Figure 5 shows the results of disease activity and H&E staining after oral administration of the microvesicles of the present invention to a mouse model.

[0023] Figure 6 shows an IVM-CM image.

[0024] Figure 7 shows the results of an in vivo imaging experiment conducted after optical labeling of microvesicles for biomobility analysis.

[0025] The present inventors have confirmed that microvesicles extracted and isolated from Bifidobacterium longumsubsp.infantis by culturing it under anaerobic conditions have a therapeutic effect on inflammatory diseases, and have completed and provided an invention. In the present invention, a pharmaceutical composition for the prevention or treatment of inflammatory diseases is provided, which comprises microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0026] Exosomes, microvesicles, or extracellular vesicles are small, nano-sized (30–150 nm) vesicles secreted by most cells. The interior of exosomes and their phospholipid bilayer membranes are known to contain various cell-derived proteins, genetic material (DNA, mRNA, miRNA), lipids, and more. Furthermore, tissue-derived exosomes have been reported to reflect the condition of the tissue from which they were secreted, possess the cytobiological characteristics of the original cell, or contain secretory peptides from the original cell, and thus can be utilized in the diagnosis and treatment of diseases.

[0027] In the present invention, the Bifidobacterium longumsubsp.infantis refers to the Infantis subspecies of the Longum species of the genus Bifidobacterium, which is an intestinal anaerobic bacterium, and may preferably have 16s rRNA represented by sequence number 1.

[0028] In the present invention, the term "prevention" means any act of inhibiting or delaying the occurrence, spread or recurrence of cancer by administering the composition of the present invention, and "treatment" means any act of improving or beneficially changing the symptoms of the disease by administering the composition of the present invention.

[0029] The term "pharmaceutical composition" in the present invention refers to a composition manufactured for the purpose of preventing or treating the above-mentioned disease, and may be formulated and used in various forms according to conventional methods. For example, it may be formulated in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and may be formulated and used in the form of topical preparations, suppositories, and sterile injectable solutions.

[0030] In the present invention, "including as an active ingredient" means that the ingredient is included in an amount necessary or sufficient to realize a desired biological effect. In actual application, the amount included as an active ingredient can be determined by considering the amount for treating the target disease and not causing other toxicity, and may vary depending on various factors such as the disease or condition being treated, the form of the composition being administered, the size of the subject, or the severity of the disease or condition. A person of ordinary skill in the art to which the present invention pertains can empirically determine the effective amount of an individual composition without undue experimentation.

[0031] In addition, the pharmaceutical composition of the present invention may, depending on each formulation, additionally include one or more pharmaceutically acceptable carriers in addition to the above-described effective ingredients.

[0032] The pharmaceutically acceptable carrier may be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components, and may further include other conventional additives such as antioxidants, buffers, and bacteriostatic agents, if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into an injectable formulation such as an aqueous solution, suspension, or emulsion, or into a pill, capsule, granule, or tablet. Furthermore, the composition may be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA).

[0033] The composition of the present invention can be administered orally or parenterally in a pharmaceutically effective amount depending on the intended method, and the term “pharmaceutically effective amount” of the present invention means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined based on factors including the patient’s health condition, severity, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, drugs used in combination or simultaneously, and other factors well known in the medical field.

[0034]

[0035] In addition, the present invention can provide a method for preventing or treating an inflammatory disease, comprising a step of administering microvesicles derived from Bifidobacterium longumsubsp.infantis to a subject.

[0036] In the present invention, the term “subject” is not limited to a mammal such as a livestock or a human that requires prevention, treatment, and / or diagnosis of the disease, but may preferably be a human.

[0037] The term "administration" in the present invention means providing a predetermined substance to a patient by any suitable method, and the pharmaceutical composition of the present invention can be formulated in various forms for administration to a subject, and a representative example of a formulation for parenteral administration is an injectable formulation, preferably an isotonic aqueous solution or suspension. The injectable formulation can be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component can be dissolved in saline or a buffer solution to be formulated for injection. In addition, formulations for oral administration include, for example, ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers, and these formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine) and a lubricant (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). The above tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and optionally may further contain a disintegrating agent such as starch, agar, alginic acid or its sodium salt, an absorbent, a coloring agent, a flavoring agent and / or a sweetening agent. The above formulations may be prepared by conventional mixing, granulating or coating methods.

[0038] In addition, the pharmaceutical composition of the present invention may further include auxiliary agents such as preservatives, wetting agents, emulsifying agents, salts for osmotic pressure control or buffers, and other therapeutically useful substances, and may be formulated according to conventional methods.

[0039] The pharmaceutical composition according to the present invention can be administered via various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular. The dosage of the active ingredient can be appropriately selected based on various factors, such as the route of administration, the patient's age, sex, weight, and severity of the condition. Furthermore, the composition of the present invention can be administered in combination with known compounds capable of enhancing the desired effect.

[0040] The pharmaceutical composition according to the present invention may be administered to humans or animals orally or parenterally, such as intravenously, subcutaneously, intranasally, or intraperitoneally. Oral administration also includes sublingual administration. Parenteral administration includes injection methods such as subcutaneous injection, intramuscular injection, and intravenous injection, as well as drip methods.

[0041] In the pharmaceutical composition of the present invention, the total effective amount of the microvesicles derived from Bifidobacterium longumsubsp.infantis according to the present invention can be administered to a patient as a single dose, or can be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the effective ingredient depending on the severity of the disease, but can typically be administered several times a day in an effective dose of 100 μg to 3,000 mg per administration for adults, but is not limited to the content.

[0042] In addition, the pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route, and administration method as long as it exhibits the effects of the present invention, and the pharmaceutical composition of the present invention may additionally include a known drug in addition to the microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases.

[0043] According to one side, the microvesicles may have an average diameter of 50 to 150 nm.

[0044] According to one side, the microvesicles may be naturally or artificially secreted from Bifidobacterium longumsubsp.infantis.

[0045] According to one aspect, the microvesicles may be isolated from a culture solution of Bifidobacterium longumsubsp.infantis or from food cultured with the addition of Bifidobacterium longumsubsp.infantis.

[0046] According to one aspect, the inflammatory disease may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease, and inflammatory bowel disease.

[0047] According to another embodiment of the present invention, a food composition for preventing or improving an inflammatory disease is provided, comprising microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

[0048] In the present invention, the term "improvement" means any action that beneficially changes a parameter related to the condition being treated, for example, at least reducing the severity of symptoms, or improving the disease.

[0049] The above food composition contains, for example, microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient, and when using it as an additive of the food composition, it can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. Generally, when manufacturing a food or beverage, the composition of the present invention is added in an amount of 15 wt% or less, preferably 10 wt% or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient can be used in an amount above the above range. That is, the mixing amount of the active ingredient can be appropriately determined depending on each purpose of use, such as prevention, health, or treatment.

[0050] The formulation of the above food composition may be in the form of powder, granules, pills, tablets, capsules, or any other form of general food or beverage.

[0051] The food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. Specifically, the food can include proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents, and examples of the carbohydrates include, but are not limited to, glucose, fructose, maltose, sucrose, oligosaccharides, dextrin, cyclodextrin, xylitol, sorbitol, erythritol, saccharin, or synthetic flavoring agents.

[0052] According to another embodiment of the present invention, a method for producing microvesicles derived from Bifidobacterium longumsubsp. infantis is provided, comprising: 1) culturing Bifidobacterium longumsubsp. infantis under anaerobic conditions to obtain a culture solution; 2) centrifuging the culture solution; and 3) separating microvesicles through tangential flow filtration (TFF).

[0053] According to another specific example of the present invention, microvesicles derived from Bifidobacterium longumsubsp.infantis, manufactured by the above manufacturing method, are provided.

[0054]

[0055] The terms used in the examples are for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0056] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0057] The present invention is susceptible to various modifications and embodiments. Specific embodiments are illustrated in the drawings and described in detail in the following detailed description. However, this is not intended to limit the present invention to specific embodiments, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. In describing the present invention, detailed descriptions of related known technologies will be omitted if they are deemed to obscure the gist of the present invention.

[0058] Example 1. Establishment of culture conditions for beneficial intestinal bacteria and isolation of nanovesicles.

[0059] In order to establish the conditions for culturing anaerobic beneficial intestinal bacteria for the discovery of therapeutic substances for inflammatory bowel disease, the following experiment was conducted. An anaerobic gas pack (AnaerogGen, Thermo Scientific), which creates an anaerobic environment by generating CO2 and absorbing oxygen in the anaerobic jar, was placed in an anaerobic jar to establish the conditions for culturing anaerobic beneficial intestinal bacteria. Using the established anaerobic culture conditions, Bifidobacterium longumsubsp. infantis was cultured for 48 hours in MRS broth containing 0.05% w / v L-cysteine. The biological resource used in the present invention was purchased from the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology (Collection number KCTC 3249 (=KCTC 3270) and is identical to ATCC 15697 and JCM 1222). The 16s rRNA sequence of the biological resource is as shown in SEQ ID NO: 1. The isolation technique for nanovesicles derived from anaerobic intestinal beneficial bacteria was optimized as follows. Afterwards, the Bifidobacterium longumsubsp. infantis culture medium was centrifuged at 6000 rpm at 4℃ for 20 minutes to remove bacterial debris and filtered once more through a 0.22 um filter. Nanovesicles derived from Bifidobacterium longumsubsp. infantis (B. infantis-EV) were isolated and concentrated from the filtered culture medium using a Tangential Flow Filtration System (TFF, Vivaflow 50, 100K MWCO). Through the above separation process, compared to the existing nanovesicle isolation method using ultra-high-speed centrifugation and filtration concentration, the extraction time was shortened and B. infantis-EV production with high yield, high purity, and high uniformity was possible. A schematic diagram of the anaerobic vessel is shown in Fig. 1A, the culture process is shown in Fig. 1B, and a photograph of the culture device is shown in Fig. 1C.

[0060] To analyze the characteristics of nanovesicles derived from beneficial gut bacteria, the following experiments were conducted. The size distribution of the extracted nanovesicles derived from beneficial gut bacteria was analyzed and evaluated through nanoparticle tracking analysis (NTA). The results are shown in Fig. 2A. The number and size of nanovesicles derived from Bifidobacterium longumsubsp.infantis were measured by NTA. The average diameter of B. infantis-EV was 73.8 nm, and the number was measured to be 3.96e+10 particles / mL.

[0061] The extracted intestinal beneficial bacteria-derived nanovesicles were fixed by adsorption onto a grid for transmission electron microscopy, stained with uranyl acetate, and the size and shape of the nanovesicles were confirmed using a transmission electron microscope (TEM), and the results are shown in Figure 2B.

[0062] The extracted intestinal beneficial bacteria nanovesicles were fixed on a poly-L-lysine cover glass and coated with gold. The size and shape of the nanovesicles were confirmed using a scanning electron microscope (SEM), and this is shown in Figure 2C.

[0063]

[0064] Example 2. Evaluation of the anti-inflammatory efficacy of B. infantis-EV

[0065] In order to evaluate the anti-inflammatory effect of B. infantis-EV cultured and isolated through the above Example 1 on the colon cancer epithelial cell line, 1 x 10 colon cancer epithelial cell line HT29 cells 6After treating dogs with 100 ng / mL of LPS and B. infantis-EV for 24 hours, the expression levels of inflammatory cytokine mRNA were confirmed through qRT-PCR, and the results are shown in Fig. 3. Compared to the control group, the group treated with B. infantis-EV showed a decrease in the expression of inflammatory cytokine mRNA, including IL-1β, IL-2, and IL-6.

[0066] Likewise, to confirm the anti-inflammatory effect of B. infantis-EV through regulation of macrophage polarization, the following experiment was conducted. The mouse macrophage cell line Raw 264.7 was differentiated into M1 macrophages by treating it with 100 ng / mL of LPS and 10 ng / mL of IFN-γ, and at the same time, B. infantis-EV was treated. The phenotype and inflammatory cytokine expression were analyzed, and the results are shown in Fig. 4. Fig. 4A is a micrograph showing the change in macrophage phenotype. Fig. 4B shows the expression of M1 macrophage markers, and Fig. 4C shows the expression of inflammatory cytokines. When observing the change in phenotype, a decrease in the M1 macrophage phenotype was confirmed in the B. infantis-EV treated group, and a decrease in the expression of M1 macrophage markers such as CD86 and INOS, as well as a decrease in the expression of inflammatory cytokines such as IL-1β, was confirmed. This is because B. infantis-EVs inhibit M1 macrophage differentiation and may be involved in anti-inflammatory action.

[0067] Similarly, to confirm the anti-inflammatory effect of B. infantis-EV in the DSS-induced acute enteritis mouse model, the following experiment was conducted. A DSS-induced acute enteritis mouse model was established by dissolving 3% DSS in drinking water and administering it for 5 days. 1x10 B. infantis-EV was injected into the mouse model. 10After oral gavage administration of the particles a total of 5 times, the disease activity index (DAI) was checked and H&E analysis was performed. The results are shown in Fig. 5. Compared to the DSS control group, the group administered B. infantis-EV showed the most significant increase in intestinal length, decrease in disease activity index (DAI), and increase in body weight (Fig. 5 B, C, and D). This confirmed the excellent anti-inflammatory effect of B. infantis-EV in the DSS-induced acute enteritis mouse model.

[0068]

[0069] Example 3. Biomobility analysis

[0070] To analyze the biotransport of B. infantis-EVs in a DSS-induced acute enterocolitis mouse model using in vivo imaging, the following experiments were conducted. B. infantis-EVs were labeled with a fluorescent marker (Cy5) and orally administered to a DSS-induced acute enterocolitis mouse model. Twenty-four hours after administration, the EVs were analyzed using Intravital Confocal & Two Photon Convertible Microscopy (IVM-CM), which combines confocal microscopy and two-photon convertible microscopy to enable real-time, high-resolution, three-dimensional imaging of cellular changes within tissues in a living mouse model. The corresponding images are shown in Fig. 6. Compared to the control mice, B. infantis-EVs were more highly expressed in the colon crypts of the DSS-induced acute enterocolitis mouse model. This demonstrates that B. infantis-EVs have a targeting preference for inflammatory bowel disease lesions.

[0071] B. infantis-EVs labeled with a photolabel (Cy5) were orally administered to a DSS-induced acute enteritis mouse model and observed using a night owl in vivo imaging device after 0, 1, 3, 8, 24, and 48 hours. The results are shown in Fig. 7. In the DSS-induced acute enteritis mouse model, B. infantis-EVs remained in the mouse body for approximately 24 hours, and compared to the control group, they were confirmed to remain in the intestine for a longer period of time in the B. infantis-EV group.

[0072]

[0073] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0074] Therefore, other implementations, other manufacturing examples and equivalents to the patent claims also fall within the scope of the claims described below.

Claims

1. A pharmaceutical composition for preventing or treating inflammatory diseases, comprising microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient.

2. In paragraph 1, A pharmaceutical composition wherein the microvesicles have an average diameter of 50 to 150 nm.

3. In paragraph 1, A pharmaceutical composition wherein the above microvesicles are naturally or artificially secreted from Bifidobacterium longumsubsp.infantis.

4. In paragraph 1, A pharmaceutical composition characterized in that the above microvesicles are isolated from a culture solution of Bifidobacterium longumsubsp.infantis or from a food cultured with Bifidobacterium longumsubsp.infantis.

5. In paragraph 1, A pharmaceutical composition, wherein the inflammatory disease is at least one selected from the group consisting of ulcerative colitis, Crohn's disease, intestinal Behcet's disease, indeterminate colitis, chronic gastritis, chronic peptic ulcer, celiac disease, and inflammatory bowel disease.

6. A food composition for preventing or improving inflammatory bowel disease, comprising microvesicles derived from Bifidobacterium longumsubsp.infantis as an active ingredient. 7.1) A step of culturing Bifidobacterium longumsubsp.infantis under anaerobic conditions to obtain a culture solution; 2) a step of centrifuging the culture solution; and 3) A method for producing microvesicles derived from Bifidobacterium longumsubsp.infantis, comprising: a step of separating microvesicles through tangential flow filtration (TFF); 8. In paragraph 7, Microvesicles derived from Bifidobacterium longumsubsp.infantis, manufactured by the above manufacturing method.

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