KR101507519B1 - Compositon for inducing maturation of dendritic cell comprising MAB2560 - Google Patents

Abstract

The present invention relates to a dendritic cell maturation inducing composition comprising an MAB2560 protein derived from Mycobacterium abscessus as an active ingredient, and a method of using the same to differentiate immature dendritic cells into mature dendritic cells. The MAB2560 protein according to the present invention can effectively enhance the immune response of the body by effectively matured immature dendritic cells.

Description

TECHNICAL FIELD The present invention relates to a composition for inducing maturation of dendritic cells comprising MAB2560 protein as an active ingredient,

The invention Mycobacterium revenue pressure switch (Mycobacterium The present invention relates to a composition for inducing the maturation of dendritic cells comprising an MAB2560 protein derived from E. abscessus as an active ingredient and a method for differentiating immature dendritic cells into mature dendritic cells using the composition.

Dendritic cells or dendritic cells (DCs) are immune cells that form part of the immune system of mammals. These cells act as antigen-expressing cells that make the other cells of the immune system recognize them by treating the antigenic material and making it appear on the surface. Dendritic cells are present in small amounts in tissues that are in contact with the external environment, such as skin, nose, lung, stomach and intestinal wall, and the cells in the skin are called Langerhans cells. Dendritic cells can be found immature in blood, and when activated, they migrate to the lymphatic organs and interact with T cells and B cells to initiate an immune response. At certain developmental stages, they stretch the dendrites, called dendrites.

Dendritic cells are derived from hemopoietic bone marrow progenitor cells. These progenitor cells are initially transformed into immature dendritic cells, characterized by high endocytosis activity and T-cell activation capacity. Immature dendritic cells constantly digest surrounding pathogens such as viruses and bacteria. This is possible through pattern recognition receptors (PRRs) such as toll-like receptors (TLRs). TLRs recognize specific chemical characteristics found on a subset of pathogens, and immature dendritic cells digest the cell membrane through a process known as nibbling from living autologous cells. When they come into contact with existing antigens, they become activated to mature dendritic cells and migrate to the lymph nodes. Immature dendritic cells digest pathogens and break down their proteins into small pieces that, when mature, appear on the cell surface using MHC (Major Histocompatibility Complex) molecules. At the same time, it increases cell surface receptors that act as co-receptors in T-cell activation such as CD (Cluster of Differentiation) 80, CD86, and CD40. They activate helper T-cells, killer T-cells as well as B-cells by showing pathogen-derived antigens with specific non-antigenic co-stimulatory signals.

All helper T-cells are specific for one particular antigen. Only specialized antigen-expressing cells (macrophages, B lymphocytes and dendritic cells) activate the remaining helper T-cells when the correct antigen is present. However, macrophages and B lymphocytes can only activate memory T-cells, while dendritic cells are the most potent antigen-expressing cells because they can activate both memory and naive T-cells.

Mature dendritic cells can originate from monocytes, the circulating white blood cells of the body. Monocytes can be turned into dendritic cells as well as dendritic cells, depending on the appropriate signal. Monocytes originate from the bone marrow stem cells. Monocyte-derived dendritic cells can be generated from peripheral blood mononuclear cells (PBMC) in the laboratory. PBMCs can be implanted in tissue culture flasks to attach monocytes, which can be differentiated into immature dendritic cells by treatment with IL-4 and GM-CSF (granulocyte-macrophage colony stimulating factor). Subsequent treatment with TNF- will differentiate immature dendritic cells into mature dendritic cells.

Fully mature DCs differ qualitatively and quantitatively from immature DCs. Fully mature DCs express high levels of MHC (Major Histocompatibility Complex) type I and type II antigens, and high levels of T cell co-stimulatory molecules, CD80 and CD86. These changes increase the T cell activation capacity of dendritic cells, which not only increases the antigen density on the cell surface, but also increases the amount of T cell activation through the counterparts of co-stimulatory molecules on T cells, such as CD28 . In addition, mature DCs produce large amounts of cytokines, which stimulate and induce T cell responses. Two of these cytokines are interleukin 10 (IL-10) and interleukin 12 (IL-12). These cytokines have the opposite effect on the direction of the induced T cell response. When IL-10 is produced, a Th-2 type reaction is induced, whereas when IL-12 is produced, a Th-1 type reaction is induced. The latter response is particularly preferred when a cellular immune response is required, for example in cancer immunotherapy. The Th-1 type response induces and polarizes cytotoxic T lymphocytes (CTLs), which are the means of attacking the working factors of the cell's immune system. This means of action factor attack is most effective in inhibiting tumor growth. IL-12 also induces the growth of natural killer (NK) cells and has anti-angiogenic activity, which are all effective anti-tumor attack means. Thus, the use of dendritic cells to produce IL-12 is theoretically optimal for use in immunostimulation.

Since dendritic cells are rare and difficult to isolate, the precise production and development of dendritic cells of different types and subsets and their interrelationships have only been briefly known. Dendritic cells communicate with other cells in the body. Such communication can take the form of direct cell-to-cell contact based on the interaction of cell surface proteins.

The cytokines produced by dendritic cells differ depending on the type of cell. Lymphoid dendritic cells can produce large amounts of type 1 IFN, which aggregates more activated macrophages and allows phagocytosis. Lymphoid dendritic cells are involved in central and peripheral immunoregulation, and myeloid dendritic cells are known to be involved in immune induction against adventitious antigens or infections. Thus, if dendritic cells fail to function normally, autoimmune diseases such as diabetes, rheumatoid arthritis, allergic hypersensitivity, or normal immune response against infectious diseases or cancer will not occur.

In cancer patients, the function of dendritic cells is deteriorated and it is difficult to induce normal cancer immunity. (LPS, TNF-, and IL-1), which regulate (activate) dendritic cell differentiation, are known to be a side effect of bio-toxicity.

On the other hand, Mycobacterium contains not only species that cause serious diseases such as tuberculosis, tuberculosis and leprosy, but also bacteria called opportunistic infectious microorganisms, About 72 species have been known to date, including saprophytic species, of which 25 are known to be related to human disease.

Mycobacterium abscessus is a bacterium widely found in nature such as soil and water. It is the only mycobacteria resistant to all antituberculous drugs currently in use. In the past, people with impaired immune function were recognized as opportunistic infectious agents causing disease in humans. Recently, however, they have been recognized as antigens causing serious diseases in humans regardless of immune function. The genome of the Mycobacterium infosus (ATCC19977) is about 5 Mb in size, and unlike other mycobacteria, it does not have a characteristic insertion sequence but has a full-length prophage of 81 kb. Mycobacterium infosus is known to be similar to M. smegmatis, which has relatively conserved genes compared to M. tuberculosis and has no pathogenicity.

As described above, dendritic cells play an important role in enhancing the immune function of the body itself. Therefore, the development of a non-toxic immunomodulatory agent that induces a strong immune response by promoting and matured dendritic cell differentiation and a clear understanding of its mechanism of action Is an important task for cell-mediated immunotherapy using dendritic cells

The inventors of the present invention have completed the present invention by discovering that the MAB2560 protein derived from Mycobacterium abscessus effectively matures dendritic cells.

It is an object of the present invention to provide a composition for inducing maturation of immature dendritic cells comprising MAB2560 protein as an active ingredient.

It is yet another object of the present invention to provide a method for inducing maturation of immature dendritic cells comprising treating mature dendritic cells with MAB2560 to mature dendritic cells.

It is still another object of the present invention to provide a composition for enhancing immunity comprising the composition for inducing dendritic cell maturation.

It is an object of the present invention to provide a composition for inducing maturation of immature dendritic cells comprising MAB2560 protein as an active ingredient.

It is yet another object of the present invention to provide a method for inducing maturation of immature dendritic cells comprising treating mature dendritic cells with MAB2560 to mature dendritic cells.

It is still another object of the present invention to provide a composition for enhancing immunity comprising the composition for inducing dendritic cell maturation.

MAB2560 according to the present invention promotes the differentiation of immature dendritic cells, thereby effectively matured dendritic cells, thereby effectively enhancing the immune response of the body.

Figure 1 shows a recombinant protein MAB2560 isolated by cloning.
Fig. 2 is a graph showing the results of an experiment for measuring the survival rate of dendritic cells treated with MAB2560.
FIG. 3 is a graph showing the expression levels of CD80, CD86, MHC class I, and II, which are specific protein surface factors in dendritic cells treated with MAB2560.
FIG. 4 shows secretion amounts of cytokines (TNF-, IL-6, IL-1, IL-12 p70, IL-10) in dendritic cells treated with MAB2560.
FIG. 5 is a graph showing the results of measurement of dextran-FITC phagocytosis of dendritic cells treated with MAB2560.
FIG. 6 is a graph showing the results of measuring the activity of T cell stimulation of dendritic cells treated with MAB2560 and the secretion amount of cytokine of T cells. FIG.
FIG. 7 shows changes in signal transduction system in dendritic cells treated with MAB2560. FIG.

The present invention provides a composition for inducing maturation of dendritic cells comprising MAB2560 protein as an active ingredient.

The MAB2560 is derived from M. bovis, preferably a protein having the amino acid sequence shown in SEQ ID NO: 1, a protein encoded by the nucleotide sequence shown in SEQ ID NO: 2, and a functional equivalent of the protein . "Functional equivalent" means at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably at least 70% or more, more preferably 90% or more, Refers to a protein having a homology of at least 95% and exhibiting substantially the same physiological activity as the protein represented by SEQ ID NO: 1.

The MAB2560 protein of the present invention, as well as its amino acid sequence variants, are also within the scope of the present invention. A variant of the MAB2560 protein means a protein having a sequence that differs by deletion, insertion, non-conservative or conservative substitution, or a combination thereof, with the MAB2560 native amino acid sequence and one or more amino acid residues. Amino acid exchanges in proteins and peptides that do not globally alter the activity of the molecule are known in the art. The MAB2560 protein or variant thereof may be prepared by natural extraction or by a recombinant method based on the synthetic sequence (Merrifleld, J. Amer. Chem. Soc. 85: 2149-2156, 1963) or DNA sequence (Sambrook et al., Molecular Cloning, Cold Spring Harbor Laboratory Press, New York, USA, Second Edition, 1989).

The present invention can induce dendritic cell maturation by isolating and purifying only the MAB2560 portion of the entire genes of M. tuberculosis isolates using an E. coli expression system.

Immature dendritic cells mature to form mature dendritic cells. Adult dendritic cells lose the ability to capture up-regulated expression of cell surface molecules and various cytokines that capture and co-stimulate the antigen. In particular, mature dendritic cells express MHC type I and type II antigens at higher levels than immature dendritic cells and regulate Cluster of Differentiation (CD) 80+, CD83 +, CD86 + and CD14-. Induces an increase in antigenic density on the dendritic cell surface with more MHC expression, whereas upregulation of the co-stimulatory molecules CD80 and CD86 leads to T cell activation through co-stimulatory molecular counterparts such as CD28 on T cells Enhance the signal.

The present invention can facilitate differentiation into mature dendritic cells through a composition comprising MAB2560. The MAB2560 of the present invention may be included in the composition at a concentration of from 0.1 μg / ml to 100 μg / ml, though not limited thereto.

The present invention also provides a method for inducing maturation of immature dendritic cells comprising treating immature dendritic cells with MAB2560 to matured into mature dendritic cells.

The MAB2560 protein of the present invention can be treated at a concentration of 0.1 [mu] g / ml to 100 [mu] g / ml, but not immature dendritic cells.

The present invention can cultivate cells treated with MAB2560 to induce maturation by treating MAB2560 with immature dendritic cells. For induction of differentiation into suitable mature dendritic cells, the cells may be cultured for 12-36 hours to mature, but are not limited thereto.

Matured dendritic cells promoted by the MAB2560 protein treatment in the present invention can express general characteristics of mature dendritic cells. That is, according to the present invention, immature dendritic cells are differentiated into mature dendritic cells by treating MAB2560 with immature dendritic cells, and more preferably TNF-α, IL-12p70, IL-6 and IL- Can be converted to increased mature dendritic cells.

The present invention also provides a composition for inducing the maturation of dendritic cells comprising MAB2560 protein as an active ingredient.

According to the present invention, immature dendritic cells are treated with MAB2560 to mature immature dendritic cells, and dendritic cell maturation can increase the expression of surface protein markers. More specifically, immature dendritic cells can be treated with MAB2560 to mature dendritic cells and consequently to induce T cell activity, thereby increasing the immune response. Accordingly, the MAB2560 of the present invention can be usefully used in medicines for enhancing immunity and health functional foods.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. The pharmaceutical composition according to the present invention may be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or other oral formulations, external preparation, suppository and sterilized injection solution, . Suitable formulations known in the art are preferably those disclosed in Remington ' s Pharmaceutical Sciences, Mack Publishing Company, Easton PA.

Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations are prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, It is prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and suspension include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For a desired effect, the composition of the present invention can be administered at a daily dose of 0.001 to 1000 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way. The dosage may vary depending on factors such as the type of disease of the individual, the severity, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, Can be determined.

The pharmaceutical composition of the present invention may be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

When the composition of the present invention is used as a food additive, the composition can be added as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the composition of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The maturation of dendritic cells can be monitored by methods known in the art and cell surface markers can be detected by assays familiar to the art, such as flow cytometry and immunohistochemistry. The cells can also be monitored for cytokine production (e.g., ELISA, FACS, and other immunoassays).

Hereinafter, the present invention will be described in more detail by way of examples. It should be noted, however, that the following examples are provided to further illustrate the present invention and are not intended to limit the scope of the present invention.

Example  1. Isolation and recombination of dendritic cells MAB2560 of Cloning

1. Isolation and induction of dendritic cells

Femoral bone marrow was collected from 6 to 8 weeks old C57BL / 6 mice using bone marrow injection. The collected bone marrow was washed and the red blood cells were removed with ammonium chloride. The cells were resuspended in RPMI 1640 (10% FBS (Fetal bovine serum, calf serum), 2 mM L-glutamine, 100 U / ml penicillin / streptomycin, 50 μM mercaptoethanol, 0.1 mM nonessential amino acid, 1 mM sodium pyruvate , 20 ng / ml GM-CSF, 20 ng / ml IL-4) medium at 37 ° C and 5% CO ₂ for 8 days. GM-CSF and IL-4 were used to induce differentiation into dendritic cells.

1.2 Recombination MAB2560 of Cloning

The MAB2560 region was amplified by PCR using the genomic DNA of Mycobacterium abscessus (ATCC19977) as a template. 5'-CAT ATG ACG AGC GTT GAC GTG CCC TCG-3 'as the forward primer and 5'-AAG CTT TAG GCC GGA GGG CAC GAC CTT-3' as the reverse primer. The isolated PCR product was digested with NdeI and HindIII enzymes and inserted into the pET-28a vector, an expression vector. E. coli BL21 transformed with the pET-28a vector into which the MAB2560 gene was inserted was cultured in LB medium (containing 100 μg / ml ampicillin) at 37 ° C for 12 hours. Thereafter, 1 mM isopropyl-D-thiogalactopyranoside (IPTG) was added and incubated for 6 hours and the cells were lysed with lysis buffer (1 M DTT, lysozyme, PMSF). The recombinant protein was purified using nickel-nitrilotriacetic acid (Ni-NTA, Invitrogen, Carlsbad, CA, USA) agarose according to the manufacturer's method. The final purified recombinant protein was identified by SDS-PAGE analysis.

The results are shown in Fig.

As shown in Figure 1, recombinant MAB2560 protein appeared at 26 kDa position.

Example  2. MAB2560 Cytotoxicity

To confirm the effect of MAB2560 on the survival of dendritic cells, dendritic cells were treated with MAB2560 at a concentration of 1, 2.5 μg / ml or LPS (lipopolysaccharide) at a concentration of 50 ng / ml as a control for 24 hours. Then, double staining with propidium iodide (PI) and annexin-V was performed and analyzed by flow cytometry.

The results are shown in Fig.

As shown in FIG. 2, it was confirmed that MAB2560 at a concentration of 2.5 μg / ml did not show toxicity in dendritic cells.

Example  3. MAB2560 Of surface expression of dendritic cells

In order to confirm the effect of MAB2560 on the maturation of dendritic cells, the following experiment was conducted. The cells were maintained in OptiMEM medium containing GM-CSF and IL-4 to homogenize the maturation of all cells. Then, 2.5 μg / ml MAB2560 or 50 ng / ml LPS was added to dendritic cells After the time treatment, dendritic cells were recovered. LPS is a well known substance that promotes the maturation of dendritic cells. To analyze the expression of surface factors associated with dendritic cell maturation, staining was performed using a cell surface factor antibody such as anti-CD80, anti-CD86, anti-MHC I and II, and analyzed using a flow cytometer FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA).

The results are shown in FIG.

As shown in FIG. 3, MAB2560 increased expression of CD80, CD86, MHC class I, and MHC class II, which are protein surface factors of mature dendritic cells.

Example  4. MAB2560 On the secretion of cytokines from dendritic cells

To determine the effect of MAB2560 on dendritic cell cytokine secretion, dendritic cells were injected with 2.5 μg / ml MAB2560 or 50 ng / ml as a control group LPS was treated and cultured for 24 hours. The supernatant was then applied to an ELISA kit for TNF-α, IL-6, IL-1β, IL-10 and IL-12 p70 to determine cytokine secretion. IL-12 p70 and IL-10 are major cytokines secreted from dendritic cells, and play important roles in differentiating T cells into Th1 and Th2, respectively. IL-12 is a typical Th1-type cytokine, and IL-10 is known to inhibit the activity of activated monocytes, NK cells, and cytokines (including IL-12 p70) produced by Th1 cells.

The results are shown in Fig.

As shown in FIG. 4, MAB2560 increased expression of TNF-α, IL-6 and IL-1β, which are proinflammatory cytokines in dendritic cells, and increased expression of IL-12 p70.

Example  5. MAB2560 On by Dendritic  Analysis of antigen acquisition ability

Immature dendritic cells are good for antigen acquisition, but mature dendritic cells are known to decrease antigen acquisition capacity. Therefore, an experiment was conducted to confirm the effect of MAB2560 protein on dendritic cell endocytic activity. Immature dendritic cells were treated with 2.5 μg / ml of MAB2560 or 50 ng / ml of LPS as a control and cultured for 24 hours. Fluorescein-conjugated dextran (molecular weight 40,000; Molecular Probes, Eugene, OR) was added to the cultured dendritic cells at 1 / / ml for 1 hour. After incubation at 37 ° C and 4 ° C for 30 minutes, the cells were washed with cold staining buffer to stop the reaction. After that, the cells were stained with PE (Phycoerythrin) -binding CD11c (BD bioscience), and dextran-FITC (Fluorescein isothiocyanate) was assayed using a flow cytometer.

The results are shown in Fig.

As shown in Fig. 5, it was confirmed that the dendritic (antigen) predoporative ability of dendritic cells treated with MAB2560 was lower than that of dendritic cells not treated with MAB2560. These results indicate that MAB2560 matured dendritic cells in terms of function.

Example  6. MAB2560 On by Dendritic  T cell activity assay

The most important characteristic of mature dendritic cells in vivo is the induction of T lymphocyte activity. CD4 + cells isolated from the spleen of OT-I and OT-II mice (BALB / c mice), ovalbumin transgenic mice, and mice were immunized with MAB2560 to determine if maturation-induced dendritic cells activated T lymphocytes CD8 + T cells were cultured with MAB2560-treated dendritic cells derived from C57BL / 6 mice and the degree of proliferation of T cells was measured. In addition, the secretion amount of cytokine (IFN-y) secreted from T cells was analyzed using an ELISA kit.

The results are shown in Fig.

As shown in FIG. 6, it was confirmed that when treated with MAB2560, the degree of T cell proliferation was higher than that of the control group. This suggests that MAB2560 can increase the immune activation capacity by proliferating T cells by matured dendritic cells. In addition, the secretion of IFN-γ, which is a cytokine secreted from T cells, indicates that treatment with MAB2560 biases the cells into Th1-type cells.

Example 7. Activation of dendritic cell signaling system by MAB2560

Activation of the mitogen activated protein kinases (MAPK) signaling system, such as ERK, JNK, and p38, is known to be essential for dendritic cell maturation. Therefore, immature dendritic cells were treated with 2.5 μg / ml of MAB2560 at 0 min, 5 min, 15 min, 30 min, 45 min, and 60 min, We confirmed the phosphorylation of ERK, JNK, and p38, which are related to the signal transduction system. The amount of protein was measured by a Western blotting method using a specific antibody.

The results are shown in Fig.

As shown in FIG. 7, it was confirmed that the amounts of p-p38, p-ERK and p-JNK phosphorylated by MAB2560 treatment were increased.

<110> Pusan National University Industry-University Cooperation Foundation <120> Compositon for inducing maturation of dendritic cell          MAB2560 <130> 135 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 201 <212> PRT <213> Mycobacterium abscessus <400> 1 Met Arg Thr Ile Ala Ile Arg His Arg Ala Val Ile Gly Leu Ser Ala   1 5 10 15 Val Ala Leu Ile Thr Val Gly Cys Ser Asn Gly Thr Ser Val Asp Val              20 25 30 Pro Ser Pro Glu Val Gly Leu Ile Ala Thr Thr Ser Ser Ala Ala Pro          35 40 45 Ala Gln Pro Ala Glu Val Lys Leu Ile Gly Glu Arg Asp Val Glu Val      50 55 60 Thr Leu Thr Gly Pro Ile Ala Ala Lys Tyr Ser Ser Ala Ser Glu Ser  65 70 75 80 Gln Lys Gln Ala Leu Gly Lys Pro Leu Thr Gly Asp His Asn Ala Gly                  85 90 95 Thr Arg Glu Ser Gly Ala Val Phe Gln Gln Phe Gln Gly Gly Ala Ile             100 105 110 Ile Ala Lys Asn Asn Gln Ala Gly Thr Pro Ala Phe Ile Val Val Gly         115 120 125 Lys Ile Arg Asp Ala Trp Asn Ile Gln Arg Asp Ala Asp Gly Thr Pro     130 135 140 Ser Ile Thr Gly Asn Asn Gly Ser Ala Gly Pro Leu Gly Leu Pro Thr 145 150 155 160 Ser Asp Glu Asn Thr Glu Gly Asp Gln Leu Val Ser Thr Phe Glu His                 165 170 175 Gly Lys Ile Glu Tyr Asn Ala Lys Ser Gly Glu Val Ala Val Thr Val             180 185 190 Asn Gly Lys Val Val Ser Gly Leu         195 200 <210> 2 <211> 606 <212> DNA <213> Mycobacterium abscessus <400> 2 atgaggacga tcgcaatccg ccacagggca gtcatcggcc tgagcgccgt agcgctgatc 60 actgtgggct gcagcaatgg cacgagcgtt gacgtgccct cgcccgaggt cgggctgatc 120 gccaccacct cgtccgcggc gcctgcgcag cccgccgagg tcaagctgat cggggagagg 180 gacgtcgaag tgacgctgac cggcccgatt gctgccaagt attcatcggc gagtgagagc 240 cgaagcagg ctctcggcaa gccgctgacg ggtgaccaca acgcgggtac acgcgagagt 300 ggcgcggtgt tccagcagtt ccagggtggc gccatcatcg ccaagaacaa ccaggcgggc 360 acgccggcct tcatcgtcgt gggcaagatc cgcgatgcct ggaacatcca gcgcgacgcg 420 gcggcaccc cgtcgatcac cggcaacaat ggctcggccg gcccgttggg cctgcccacc 480 agcgacgaaa acaccgaggg tgatcagctc gtgtcgacct ttgagcacgg caagatcgag 540 tacaacgcga agagcggcga ggtagcggtc acggtcaacg gcaaggtcgt gccctccggc 600 CTatag 606

Claims (8)

A composition for inducing in vitro maturation of dendritic cells isolated in vivo comprising MAB2560 protein as an active ingredient.
The composition for inducing in vitro maturation of dendritic cells isolated in vivo according to claim 1, wherein the MAB2560 protein is derived from Mycobacterium abscessus.
The composition for inducing in vitro maturation of dendritic cells separated in vivo according to claim 1, wherein the MAB2560 protein is contained at a concentration of 0.1 μg / ml to 100 μg / ml.
A method for inducing in vitro maturation of immature dendritic cells, comprising: treating immature dendritic cells isolated in vivo with MAB2560 protein to produce mature dendritic cells.
5. The method of claim 4, wherein the MAB2560 protein increases the production of TNF- [alpha], IL-12p70, IL-6 or IL-1 [beta] during the maturation of immature dendritic cells. vitro maturation induction method.
5. The method according to claim 4, wherein the MAB2560 protein is treated at a concentration of 0.1 쨉 g / ml to 100 쨉 g / ml.
A pharmaceutical composition for enhancing immunity comprising MAB2560 protein as an active ingredient.
A food composition for enhancing immunity comprising MAB2560 protein as an active ingredient.

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