KR102347797B1 - Cell surface antigen of Macrophage and the use thereof - Google Patents

Abstract

The present invention relates to a composition for diagnosing cancer comprising an agent for measuring the expression level of a CD169 (Cluster of Differentiation 169) protein or a gene encoding the same.

Description

Cell surface antigen of Macrophage and the use thereof

The present invention relates to a cell surface antigen protein specifically present on the surface of macrophages for cancer cells, particularly brain tumors, and to various uses thereof.

Cancer is one of the most common causes of death worldwide. About 10 million new cases occur each year, accounting for about 12% of all deaths, making it the third leading cause of death. Among various types of cancer, especially brain cancer occurs regardless of age, and has a higher incidence in children than other cancers. Brain cancer is divided into primary brain tumors that occur in the brain tissue and the meninges that surround the brain, and secondary brain tumors that metastasize from cancer that occurs in the skull or other parts of the body. , local symptoms such as balance disturbance, and symptoms of intracranial hypertension. The brain cancer includes various types of cancer, such as glioblastoma multiforme, glioma malignant, lymphadenoma, germ cell tumor, and metastatic tumor, unlike cancers that occur in other tissues where one or two types of cancer are tissue-specific. , among these, glioma, especially glioblastoma, is the most malignant and aggressive and has a very poor prognosis. It is a very fatal disease. In patients with glioblastoma, since the boundary between brain cells and tumor cells is not clear, it is known that it is almost impossible to completely remove cancerous tissue surgically.

Accordingly, although various studies are being conducted to develop a method for treating the glioblastoma, since glioblastoma includes various genetic mutations, the average survival rate of glioblastoma patients is remarkably high even now that the combination treatment method is used. low, the development of new therapeutic agents is required.

However, since the brain in which the glioblastoma occurs is an immune-privileged organ in which the inflow and activity of immune cells is very limited, virtually no immune response to remove tumor cells occurs, so compared to other cancers, Most of the immune cells are microglia or tumor-associated macrophages that suppress the anticancer immune response, and the infiltration of lymphocytes such as T cells is very small.

On the other hand, macrophages, which are known to exist most in tumors, are known to exist as tumor-associated macrophages, which generally suppress immune responses by the tumor environment and help tumor growth and metastasis. However, according to recent research results, tumor-associated macrophages are not composed of the same type of cell, but are composed of several subtypes with various characteristics.

Although macrophages are found in cancers such as breast cancer, liver cancer, and melanoma, their respective revealed functions and roles in tumor immune response are different. This is because macrophages expressing the same surface antigen are different cells that show different phenotypes depending on the tissue and carcinoma in which the cells are present.

According to a study analyzing human breast cancer, among macrophages inside the tumor, CD169+ macrophages secrete CCL8 to increase the malignancy of tumor cells, and it was reported that the higher the expression of CD169 in the tumor tissue, the shorter the survival period. Also, in the mouse lewis lung carcinoma model, there is a report that when CD169 macrophages are removed, T cell infiltration in the tumor increases and the size of the tumor decreases.

On the other hand, subcapsular sinus macrophages of LN phagocytose dead tumor cells that have flowed into the lymph and trigger a T-cell immune response to suppress tumor growth. In liver cancer, CD169+ macrophages inhibit the division and activity of cytotoxic T cells. It has been reported that by increasing it inhibits tumor growth.

Also, in the case of melanoma, it has been reported that both tumor-associated macrophages that suppress the anticancer immune response and general macrophages that do not express CD169.

Although all these cells have in common that they are macrophages expressing CD169, their revealed functions and roles in the tumor immune response are different. This means that even macrophages expressing the same CD169 are different cells showing different phenotypes depending on the tissue and carcinoma in which the cells are present.

Therefore, in the present invention, by confirming the characteristics and functions of macrophages in brain tumors, the role of these cells in the anti-brain tumor immune response is to be identified and used for diagnosis and treatment.

An object of the present invention is to provide a composition for diagnosing cancer comprising an agent for measuring the expression level of a CD169 (Cluster of Differentiation 169) protein or a gene encoding the same.

Another object of the present invention relates to a method for providing information for cancer diagnosis, comprising measuring the expression level of CD169 protein or a gene encoding the same in a biological sample isolated from a target individual.

Another object of the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising a macrophage comprising CD169 protein or an extracellular domain thereof as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, various specific details are set forth, such as specific forms, compositions and processes, and the like, for a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, references to "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, the particular features, forms, compositions, or properties may be combined in any suitable manner in one or more embodiments.

Unless specifically defined in the present invention, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

In the present invention, "tumor" or "cancer" is a disease in which cell division is continued due to uncontrolled cell cycle, and is divided into carcinoma (Carcinoma) and sarcoma (Sarcoma) according to the site of occurrence. Carcinoma refers to a malignant tumor that arises from epithelial cells such as mucous membranes and skin, and sarcoma refers to a malignant tumor that arises from non-epithelial cells such as muscle, connective tissue, bone, cartilage, and blood vessels.

In the present invention, the cancer is breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, thymus cancer, squamous cell carcinoma, adenocarcinoma, and combinations thereof may be selected from the group consisting of.

In the present invention, the brain cancer may be glioblastoma (GBM). The glioblastoma (GBM) is a type of intractable brain tumor, and the average total survival period after diagnosis is only about 15 months. Glioblastoma refers to tumors originating from glial cells, which are normally abundant in brain tissue.

In the present invention, "expression" refers to a process in which a polynucleotide is transcribed into mRNA and/or mRNA is translated into a peptide, polypeptide, or protein. When the polynucleotide is derived from the genomic DNA of an appropriate eukaryotic host, expression may include splicing of the mRNA. Regulatory elements necessary for expression include a promoter sequence for RNA polymerase binding and a transcription initiation sequence for ribosome binding. For example, a bacterial expression vector or cassette contains a promoter (eg, the lac promoter) and a Shine-Dalgarno sequence for transcription initiation and a start codon AUG (Sambrook et al. (1989), supra). Similarly, eukaryotic expression vectors or cassettes generally contain a heterologous or homologous promoter for RNA polymerase II, a Kozak sequence, a start codon AUG, a stop codon for ribosome exit, and a downstream polyadenylation signal. The vector can be purchased commercially or can be made with the sequence described by a method known in the art.

In the present invention, "CD169 (Cluster of Differentiation 169)" is also called siglec-1, sialoadhesin, and is a cell adhesion molecule found on the surface of macrophages. CD169 contains 17 immunoglobulin (Ig) domains (1 variable domain and 16 constant domains) and is defined as a type I lectin. The CD169 protein is encoded by the siglec-1 gene.

In the present invention, the CD169 protein may consist of the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto.

In the present invention, "CD45 (Cluster of Differentiation 45)" is also referred to as protein tyrosine phosphatase receptor type C (PTPRC), and is encoded by the PTPRC gene.

In the present invention, the CD45 protein may consist of the amino acid sequence shown in SEQ ID NO: 2, but is not limited thereto.

In the present invention, "F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1; EMR1)" is encoded by the ADGRE1 gene.

In the present invention, the F4/80 protein may consist of the amino acid sequence represented by SEQ ID NO: 3, but is not limited thereto.

In the present invention, "Ly6c (Lymphocyte antigen 6 complex)", also known as lymphocyte antigen 6 (ly6) or urokinase-type plasminogen activator receptor (uPAR), is one of a family of proteins that share a common structure but have different tissue expression patterns and functions . Ly6C is a mouse antigen, and Ly6C+ monocytes of mice are classified as classical monocytes. In humans, the classical monocyte marker for Ly6C+ monocytes is CD14.

In the present invention, the ly6c protein may consist of the amino acid sequence shown in SEQ ID NO: 4, but is not limited thereto.

In the present invention, the "CD14 (Cluster of Differentiation 14)" is a human protein mainly produced by macrophages as a part of the innate immune system. CD14 helps to detect bacteria in the body by binding to lipopolysaccharide (LPS), a pathogen-associated molecular pattern (PAMP).

In the present invention, the CD14 protein may consist of the amino acid sequence represented by SEQ ID NO: 5, but is not limited thereto.

In the present invention, Ly6G (Lymphocyte antigen 6 complex locus G6D) is a marker for classifying mouse neutrophils. Ly6G is a differentiation antigen linked to 21-25 kD glycosylphosphatidylinositol (GPI) expressed by bone marrow-derived cells in a developmentally regulated manner in the bone marrow. Monocytes transiently express Ly6G during myelogenesis, whereas Ly6G expression in granulocytes and peripheral neutrophils is directly related to the level of differentiation and maturation of cells. Human neutrophil markers may use CD15 and CD66b, but are not limited thereto.

In the present invention, CD15 (Cluster of Differentiation 15), also referred to as Lewis x, is a distinct marker for human bone marrow cells and mediates neutrophil adhesion to dendritic cells.

In the present invention, CD66b (Cluster of Differentiation 66b) is also called CEACAM8 and is a kind of CEA gene family. The main functions of the CD66b are cell adhesion, cell migration and pathogen binding.

In the present invention, "macrophage" is one of the immune cells, also called phagocytes. It is a general term for large amoeba-like phagocytes that are distributed in all tissues of the animal body, and eat and digest foreign substances, bacteria, viruses, and waste cells in the body. The macrophages of the present invention may be microglia of the central nervous system.

In the present invention, "monocytes" are a type of white blood cells derived from hematopoietic stem cells (HSCs) in the bone marrow, and are differentiated from common bone marrow progenitor cells during the differentiation process. Monocytes are large-sized white blood cells, mature in the bone marrow, circulate in blood vessels, move to tissues, differentiate into macrophages or dendritic cells of the bone marrow, and perform functions of phagocytosis, antigen presentation, and cytokine production.

In the present invention, "differentiation" is a process in which undifferentiated cells are transformed into special cells constituting specific tissues and organs, and differentiated cells have unique structures and functions, unlike undifferentiated cells. Cellular differentiation occurs in both plant and animal cells and involves changes in intracellular gene expression patterns. In general, differentiated cells retain their traits permanently, but under special conditions, they can be transformed into other types of cells.

In the present invention, "monocyte-derived" means that the differentiated cells are obtained from monocytes.

According to one embodiment of the present invention, it relates to a composition for diagnosing cancer comprising an agent for measuring the expression level of a CD169 (Cluster of Differentiation 169) protein or a gene encoding the same.

The composition for diagnosing cancer of the present invention is at least one selected from the group consisting of CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1), and CD14 (Cluster of Differentiation 14). It may further include an agent for measuring the expression level of a protein or a gene encoding the same.

The protein of the present invention or a gene encoding the same may be expressed in macrophages.

The macrophages of the present invention may be derived from monocytes.

The agent for measuring the expression level of the protein of the present invention includes at least one selected from the group consisting of an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the protein can do.

The agent for measuring the expression level of the gene encoding the protein of the present invention may include one or more selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the gene.

According to another embodiment of the present invention, it is possible to provide a kit for diagnosing cancer including a composition for diagnosing cancer including an agent for measuring the expression level of a CD169 (Cluster of Differentiation 169) protein or a gene encoding the same.

The cancer diagnostic kit of the present invention includes at least one selected from the group consisting of Cluster of Differentiation 45 (CD45), EGF-like module-containing mucin-like hormone receptor-like 1 (F4/80), and Cluster of Differentiation 14 (CD14). It may further include an agent for measuring the expression level of a protein or a gene encoding the same.

The protein of the present invention or a gene encoding the same may be expressed in macrophages.

The macrophages of the present invention may be derived from monocytes.

In the present invention, "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. In addition to direct labeling of a detectable substance with respect to a probe or antibody by reaction with a substrate, indirect labeling in which a color-generating label is conjugated by reactivity with another directly labeled reagent is included. It may include a chromogenic substrate solution, a washing solution, and other solutions to undergo a color reaction with the label, and may be prepared including reagent components used. In the present invention, the kit may be a kit including essential elements necessary for performing RT-PCR, and in addition to each primer pair specific for a marker gene, a test tube, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerization enzymes, reverse transcriptase, DNase, RNase inhibitors, sterile water, and the like. Also, the kit may be a kit including essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which cDNA corresponding to a gene or fragment thereof is attached as a probe, and the substrate may include cDNA corresponding to a quantitative control gene or fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or a multiple reaction monitoring (MRM) kit.

The kit of the present invention may further include one or more other component compositions, solutions or devices suitable for the assay method. For example, in the present invention, the kit may further include essential elements necessary for performing the reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes a pair of primers specific for a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably, a length of about 10 bp to 30 bp. It may also include a primer specific for the nucleic acid sequence of the control gene. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (with varying pH and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC -Water (DEPC-water), sterile water, etc. may be included.

In addition, the kit for diagnosing cancer of the present invention may include essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which cDNA or oligonucleotide corresponding to a gene or fragment thereof is attached, and reagents, agents, enzymes, etc. for preparing a fluorescently-labeled probe. The substrate may also contain cDNA or oligonucleotides corresponding to control genes or fragments thereof.

In addition, the kit for diagnosing cancer of the present invention may include essential elements necessary for performing ELISA. The ELISA kit contains an antibody specific for this protein. Antibodies are antibodies with high specificity and affinity for a marker protein and little cross-reactivity with other proteins, and are monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. The ELISA kit may also include an antibody specific for a control protein. Other ELISA kits include reagents capable of detecting bound antibody, for example, labeled secondary antibodies, chromophores, enzymes (eg, conjugated with an antibody) and their substrates or capable of binding the antibody. other materials and the like.

In the cancer diagnostic kit of the present invention, as a fixture for antigen-antibody binding reaction, a well plate synthesized from a nitrocellulose membrane, a PVDF membrane, a polyvinyl resin or polystyrene resin, and a glass slide Glass or the like may be used, but is not limited thereto.

In addition, in the cancer diagnostic kit of the present invention, the label of the secondary antibody is preferably a color developing agent that exhibits a color reaction. (Poly L-lysine-fluorescein isothiocyanate), RITC (rhodamine-B-isothiocyanate), such as a fluorescent substance (fluorescein) and a label such as a dye (dye) may be used, but is limited thereto no.

In addition, it is preferable to use a chromogenic substrate for inducing color development in the cancer diagnostic kit of the present invention according to a marker that undergoes a color reaction, TMB (3,3',5,5'-tetramethyl bezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD(o-phenylenediamine), etc. can be used. In this case, it is more preferable that the chromogenic substrate be provided in a dissolved state in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is decomposed by HRP used as a label of the secondary antibody conjugate to generate chromogenic deposits, and the presence or absence of the marker proteins is detected by visually confirming the degree of deposition of the chromogenic deposits.

In the cancer diagnostic kit of the present invention, the washing solution preferably contains a phosphate buffer solution, NaCl and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer, 0.13 M NaCl, and 0.05% Tween 20. This is more preferable. After the antigen-antibody binding reaction, the secondary antibody is reacted with the antigen-antibody conjugate, and an appropriate amount is added to the immobilizer and washed 3 to 6 times. As the reaction stop solution, a sulfuric acid solution (H2SO4) may be preferably used.

In another embodiment of the present invention, in a biological sample isolated from a subject of interest, CD169 (Cluster of Differentiation 169) CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor- like 1) and CD14 (Cluster of Differentiation 14) relates to a method for providing information for cancer diagnosis, comprising measuring the expression level of one or more proteins selected from the group consisting of or a gene encoding the same.

In the present invention, the biological sample refers to any material, biological fluid, tissue or cell obtained from or derived from an individual, and includes whole blood, leukocytes, and peripheral blood mononuclear cells. ), buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, respiration (breath), urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid , glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate (joint aspirate), organ secretions (organ secretions), cells (cell), cell extract (cell extract), and may be one or more selected from the group consisting of cerebrospinal fluid (cerebrospinal fluid), but is not limited thereto.

In the present invention, the step of measuring the expression level may be performed with respect to macrophages, preferably CD169+CD14+F4/80low macrophages.

In the present invention, when the CD169+CD14+F4/80low macrophages are detected during the step of measuring the expression level, the diagnosis of cancer may be an information providing method for cancer diagnosis.

In the present invention, the macrophages may be derived from monocytes, preferably monocyte-derived CD169+CD14+F4/80low macrophages.

Another embodiment of the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising a macrophage comprising CD169 protein or an extracellular domain thereof as an active ingredient.

The macrophage of the present invention is at least one selected from the group consisting of CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1), and CD14 (Cluster of Differentiation 14). It may further include a protein or an extracellular domain thereof, and preferably, the macrophages may be derived from monocytes.

In the present invention, "prevention" refers to any action that delays the progression of cancer growth, proliferation, invasiveness or metastasis by administration of the composition of the present invention.

In the present invention, "treatment" and "improvement" refer to any action that improves or beneficially changes cancer disease by inhibiting cancer growth, proliferation, invasiveness or metastasis by administration of the composition of the present invention.

In the present invention, the pharmaceutical composition may be characterized in the form of capsules, tablets, granules, injections, ointments, powders or drinks, and the pharmaceutical composition may be characterized in that it is targeted to humans.

The pharmaceutical composition of the present invention is not limited thereto, but each can be formulated in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions according to conventional methods. can The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, fragrances, etc., in the case of oral administration, and in the case of injections, buffers, preservatives, pain relief A topical agent, solubilizer, isotonic agent, stabilizer, etc. can be mixed and used, and in the case of topical administration, a base, excipient, lubricant, preservative, etc. can be used. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above. For example, in the case of oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in the form of unit dose ampoules or multiple doses. have. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained release formulation, and the like.

Meanwhile, examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used. In addition, it may further include a filler, an anti-agglomeration agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier, a preservative, and the like.

The route of administration of the pharmaceutical composition according to the present invention is not limited thereto, but oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical , sublingual or rectal. Oral or parenteral administration is preferred.

In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

The pharmaceutical composition of the present invention depends on several factors including the activity of the specific compound used, age, weight, general health, sex, diet, administration time, administration route, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated. The dosage of the pharmaceutical composition may vary depending on the patient's condition, weight, disease severity, drug form, administration route and period, but may be appropriately selected by those skilled in the art, and 0.0001 to 50 mg/day kg or 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

The composition of the present invention may be used alone or in combination with conventional anticancer agents, or may be used in combination with surgery, radiation therapy, hormone therapy, other chemotherapy and methods using biological response modifiers.

The anticancer agent of the present invention includes nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, Cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, bevacizumab, cisplatin, cetuximab, viscumalbum, asparaginase, tretinoin, hydroxycarbamide , Dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomab tuccetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan , gemcitabine, doxyfluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacitidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, enocitabine , flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmofer, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinbla Steen, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsiroli Mousse, busulfan, ifosfamide, cyclophosphamide, melparan, altretmine, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, exemestane, aminoglue 1 selected from the group consisting of tesimid, anagrelide, nabelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozol, bicalutamide, lomustine and carmustine More than one species may be used, but is not limited thereto.

Another embodiment of the present invention relates to a food composition for the prevention or treatment of cancer comprising macrophages including CD169 protein or an extracellular domain thereof as an active ingredient.

The macrophage of the present invention is at least one selected from the group consisting of CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1), and CD14 (Cluster of Differentiation 14). It may further include a protein or an extracellular domain thereof, and preferably, the macrophages may be derived from monocytes.

The food composition of the present invention may be prepared in the form of various foods, for example, beverages, gums, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, bread, and the like.

Here, when the food composition is prepared in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and it may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. That is, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, and common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol are included. can do. Examples of the flavoring agent include natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

In addition, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners , pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.

The composition for cancer diagnosis provided by the present invention relates to a preparation for measuring the expression level of a CD169 (Cluster of Differentiation 169) protein or a gene encoding the same, and particularly relates to the diagnosis of brain cancer cells.

1 is an analysis of single-cell transcriptome of macrophages infiltrating in brain tumors.
Figure 2 shows the images of CD169+ macrophages in the wild type (WT) and CD169-DTR mice in a glioblastoma model taken with a confocal microscope.
3 shows a normal brain tissue without a tumor.
4 shows the expression of CD169 in the brain tissue in which the tumor occurred.
5 shows CD169 and F4/80 expression of macrophages infiltrating tumor tissue.
6 shows the infiltration of CD169+ cells according to tumor progression.
7 is an analysis of macrophage subtypes expressing CD169 among tumor-infiltrating immune cells using flow cytometry.
8 is an analysis of single-cell transcriptome of macrophages in tumors.
9 is a graph showing the viability of CD169+ macrophages in wild-type and CD169-DTR mice in a glioblastoma model.
10 shows the total number of immune cells (CD45.2+) present in brain tumor tissue in wild-type and CD169-DTR mice.
11 shows the cell ratio and cell number of neutrophils (Ly6G+) present in brain tumor tissues in wild-type and CD169-DTR mice.
12 shows the cell ratio and cell number of monocyte-derived macrophages (Ly6G-Ly6C+F4/80+) present in brain tumor tissues in wild-type and CD169-DTR mice.
13 shows the cell ratio and cell number of bovine glial cells (Ly6G-Ly6C-F4/80+CD45.2low) present in brain tumor tissues in wild-type and CD169-DTR mice.
14 shows the cell ratio and cell number of monocytes (Ly6G-F4/80-MHCII-Ly6C+CD11b+) present in brain tumor tissues in wild-type and CD169-DTR mice.
15 shows the ratio and cell number of dendritic cells (Ly6G-F4/80-MHCII+CD11c+) present in brain tumor tissues in wild-type and CD169-DTR mice.
FIG. 16 shows the total number of immune cells (CD45.2+) present in brain tumor tissues in wild-type and CD169-DTR mice 10 days after injection ^{of 1×10 5 G1261.}
FIG. 17 shows the ratio and number of CD4+ T cells present in brain tumor tissues in wild-type and CD169-DTR mice 10 days after injection ^{of 1×10 5 G1261.}
18 shows the ratio and number of CD4+ T cells present in brain tumor tissues in wild-type and CD169-DTR mice 10 days after injection of 1×10 ^{5 G1261.}
19 shows the ratio and number of CD8 T cells present in brain tumor tissues in wild-type and CD169-DTR mice 10 days after injection of 1×10 ^{5 G1261.}
20 shows the number and ratio of regulatory T cells (CD4+Foxp3+) among intratumoral CD4+ T cells present in brain tumor tissues in wild-type and CD169-DTR mice 10 days after injection of 1×10 ^{5 G1261.}
Figure 21 shows the expression of chemokines in brain tumor tissues in wild-type and CD169-DTR mice 10 days after G126161 injection.
Figure 22 shows cytokine expression in brain tumor tissues in wild-type and CD169-DTR mice 10 days after G126161 injection.
23 shows the expression of hypoxia-related genes in brain tumor tissues in wild-type and CD169-DTR mice 10 days after G126161 injection.

Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited by the following examples.

Example

Confirmation of macrophage diversity in brain tumors

To confirm the diversity of macrophages in brain tumors, single-cell transcriptomes of infiltrating immune cells were analyzed.

Experimental Method 1

^{The GL261 brain tumor cell line of 5 Х 10 4} was injected into the brain of 8-week-old male C57BL/6 mice, and 20 days later, the right cerebral hemisphere injected with the tumor was isolated. When the brain was separated, 30 ml of DPBS was put into the heart and perfused to remove the blood in the tissue. The separated brain was sliced using a razor blade and 1% fetal bovine serum RPMI containing 2 mg/ml of Collagenase IV (Worthington Biochemical Corp) and 30 μg/ml of DNase I (Roche) Put in the media and incubated at 37 ℃ 30 minutes. The brain slices were put in a strainer (SPL, 93070) and finely ground, then put into Percoll (GE Healthcare, 17-0891-01) with a gradient of 30%/70% and centrifuged at 2000 rpm for 20 minutes to immunize. cells were obtained. The cells were stained with αCD45.2 (104) and αCD11b (M1/70) antibodies at 4°C for 30 minutes. Before reading the cells, 7-AAD was added and only CD45 hi cells were isolated by FACS ARIA.

The library was prepared using 10X genomics Chromium single-cel 3' v3 kit (10x genomics), and next generation sequencing was performed using HiSeqXten (Illumina) in Macrogen at 10,000 cells/sample depth. Raw FASTQ data were processed with Cell Ranger (10x Genomics, version 3.0.0). A matrix was created using mouse genome 10-3.0.0 (10x Genomics, inc.) as a reference genome, and transcript count and cell barcodes information were processed.

The processed data were analyzed by Seurat analysis using the R program. In order to exclude low-quality cells, samples with RNA expression levels of 200 or less or 6000 or more and samples with mitochondrial genes of 5% or more of RNA expression were removed. Each data was normalized using the NormalizeData function, and the gene expression level was confirmed using the FindVariableFeature function. Cells were clustered with a resolution of 0.6 using Findneighbors and FindClusters functions.

Experiment Result 1

Immune cells obtained from brain tumors by the above method were classified into 7 cell groups (cluster) and classified into T cells, B cells, NK/γδ T cells, neutrophils, microglia, and myeloid cells using main lineage markers. Among them, myeloid cells were analyzed again with a resolution of 0.3 and classified into 4 cell populations. As shown in Figure 1, it was confirmed that the macrophages expressing CD68 in the brain tumor consist of at least three subtypes.

Group 0 was estimated to be monocyte-derived macrophages expressing Ly6c and low-expressing F4/80, and it was confirmed that Siglec1 (CD169) was expressed in these cells. Groups 1 and 2 did not express Ly6C and CD169. In group 2, F4/80 was highly expressed. In order to confirm that these immune cells have infiltrated into the brain tumor, the immune cells of the normal brain and the tumor-generated brain were confirmed with a confocal microscope as shown in FIG. 2 .

Confirmation of the location of CD169+ macrophages in the brain and infiltrating the tumor

Experimental method 2

The G1261 cell line expressing 1 Х 10 of green fluorescent protein (GFP) was injected into the brain of 8-week-old male C57BL/6 mice, and the brain was isolated 20 days later. When separating the brain, 30 ml of DPBS was put into the heart and perfused to remove blood in the tissue, and DPBS containing 4% paraformaldehyde (PFA) was perfused to fix the tissue. Then, the brain was separated and put in a 4% PFA solution and fixed at 4°C for 16 hours. The fixed tissue was put in a DPBS solution containing 30% sugar and dehydrated at 4°C for more than 48 hours. After that, it was put in an OCT solution (Sakura, 4583) and then frozen to make a block. The prepared OCT block was cut into 40 μm thick using a cryostat (LEICA), put in DPBS containing 5% goat serum and 0.3% Triton X-100, and incubated for 1 hour at room temperature, followed by αCD169, αF4/80 or αCD31 antibody was stained with Then, slides were fabricated using a mounting solution containing DAPI (Abcam, ab104139-20ml), and immune cell infiltration was analyzed using LSM800 confocal microscopy and Zen software (ZEISS).

Experiment result 2

As shown in FIG. 3 , in normal tissues, cells expressing CD169 were present in the blood vessels, not in the actual tissues of the brain, or in the area around the blood vessels, and the number was not large. On the other hand, in the glioblastoma model, as shown in FIGS. 4 and 5 , many cells expressing CD169 were infiltrated inside the tumor formed in the brain, and these cells expressed F4/80. These cells hardly exist in normal brain tissue without cancer, and CD169-expressing cells were mainly infiltrated in the center of cancer. As shown in FIG. 6 , the intratumoral infiltration of CD169-expressing cells appeared from the early stage of tumor development and increased as the tumor grew. As a result of confirming the expression of F4/80 or CD169 inside the tumor as shown in FIG. 7 , it was found that cells expressing CD169 also expressed F4/80, but there were also cells that did not express CD169 and only expressed F4/80. This means that the macrophages in the tumor are composed of various cells rather than the same single cell.

Experimental method 3

1 Х 10 of the GL261 brain tumor cell line was injected into the brain of 8-week-old male C57BL/6 mice, and 20 days later, the tumor-injected right hemisphere was isolated. Immune cells were isolated from the brain in the same way as in Experimental Method 1. These cells were αCD45.2 (104), αF4/80 (BM8), αLy6C (AL-21), αCD11b (M1/70), αCD169 (3D6). 112) antibody was added and stained at 4°C for 30 minutes. Before reading the cells, dead cells were stained with PI and analyzed with LSR Fortessa.

Experiment result 3

When a brain without a tumor and a brain with a tumor were compared and analyzed, CD45hiF4/80+ cells appeared in the brain with a tumor, and it was found that these cells were imported from outside because they were not found in normal brain tissue without a tumor. When F4/80 and Ly6C, markers of macrophages and monocytes, were analyzed in CD45hi F4/80+ cells, they can be largely divided into F4/80hi Ly6C- and F4/80low Ly6c+ cells, and CD169 expression in F4/80low Ly6c+ cells was confirmed. could see Expression of this marker means that CD169-expressing cells in brain tumors are monocyte-derived macrophages introduced through blood. Based on the analysis results, these cells were named CD169+Ly6c+F4/80low monocyte-derived macrophages. CD169+Ly6c+F4/80low monocyte-derived macrophages can infiltrate into the parenchymal tissue of the brain when a tumor develops, and can directly affect the tumor microenvironment within the tumor. In addition, since more CD169+Ly6c+F4/80low monocyte-derived macrophages exist in the center of the cancer, the tumor microenvironment derived from hypoxia at the center of the cancer or necrosis of cancer cells is the result of CD169+Ly6c+F4/80low monocyte-derived macrophages. can be considered to trigger infiltration.

Characterization of CD169+Ly6c+F4/80low monocyte-derived macrophages

Transcript expression of CD169+Ly6c+F4/80low monocyte-derived macrophages was analyzed through single-cell transcriptome analysis.

8 is an analysis of the single-cell transcriptome of macrophages in the tumor, in macrophages (A) chemokine receptor, (B) growth factor receptor, (C) co-stimulatory factor, (D) scavenger receptor, (E ) Fcγ receptor, (F) IFN receptor, (G) T cell chemokine expression is shown.

CD169+Ly6c+F4/80low monocyte-derived macrophages highly expressed CCR2, which is important for monocyte infiltration into tissues. In all macrophage subtypes, the receptors for growth factors CSF1 and CSF2 were highly expressed. CD80, a co-stimulatory factor, was not observed in all macrophages, but CD86 was highly expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages. CD36, a scavenger receptor, is specifically expressed only in the second macrophage population, and macrophage scavenger receptor 1 (MSR1) is specifically expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages. did The Fcγ receptor mediating antibody-dependent cytotoxicity or macrophage was expressed in all macrophages, but was especially highly expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages. IFN-γ receptor 1 was highest expressed in macrophage population 2, but IFN-γ receptor 2 was highly expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages, but not in macrophage population 1. IFN-α receptor was expressed in all macrophages, but the highest expression was in CD169+Ly6c+F4/80low monocyte-derived macrophages.

CXCL9 and CXCL10, which are important chemokines for tumor invasion of T cells, were specifically and highly expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages. These results suggest that CD169+Ly6c+F4/80low monocyte-derived macrophages differentiation and tumor invasion and proliferation can be mediated by CCR2, CSF1, CSF2, and IFN. The high expression of scavenger receptors and Fcγ receptors suggests that CD169+Ly6c+F4/80low monocyte-derived macrophages have phagocytosis on tumors. In addition, since costimulatory factors important for T cell activity and chemokines important for T cell migration are highly expressed in CD169+Ly6c+F4/80low monocyte-derived macrophages, these cells mediate T cell responses important for anticancer immune response in brain tumors. can be guessed

Confirmation of the effect of CD169+Ly6c+F4/80low monocyte-derived macrophages on the anticancer immune response

According to the previous results, various macrophages exist in brain tumors, and among them, CD169+Ly6c+F4/80low monocyte-derived macrophages were expected to increase the anticancer immune response. To confirm this, CD169+Ly6c+F4/80low monocyte-derived macrophages were specifically removed to confirm the importance of CD169+Ly6c+F4/80low monocyte-derived macrophages in the tumor. CD169-DTR mice are mice in which the diphtheria toxin receptor is expressed in cells expressing CD169, and treatment with diphtheria toxin becomes a mouse capable of temporarily eliminating CD169-expressing cells. It was confirmed whether CD169+Ly6c+F4/80low monocyte-derived macrophages of brain tumors were removed in CD169-DTR mice.

Experimental method 4

1 Х 10 of the GL261-GFP brain tumor cell line was injected into the brain of 8-week-old male CD169-DTR hetero mice, and after 19 days, 1000 ng of diphtheria toxin was intraperitoneally injected. After 24 hours, the brain was isolated and sectioned in the same way as in Experimental Method 2. Sections were stained with αCD3 (SP7) and αCD169 (3D6.112) antibodies, and slides were prepared and analyzed in the same manner as in Experimental Method 2.

Experiment result 4

It was confirmed that CD169+Ly6c+F4/80low monocyte-derived macrophages disappeared from the tumor by diphtheria toxin.

Confirmation of the effect of CD169+ macrophages on the progression of glioblastoma formed in the brain

In order to confirm the function of CD169+Ly6c+F4/80low monocyte-derived macrophages in brain tumors, the survival period at the time of tumor injection was compared in mice with and without these cells as shown in FIG. 9 .

Experimental Method 5

1000 ng of diphtheria toxin was intraperitoneally injected into 8-week-old male CD169-DTR hetero or wild-type mice, and 1 Х 10 of the GL261 brain tumor cell line was injected 24 hours later. After 4 days, the same amount of diphtheria toxin was injected intraperitoneally, and the mouse survival was observed for 30 days.

Experiment result 5

In mice without CD169+Ly6c+F4/80low monocyte-derived macrophages, the survival period of mice was significantly reduced compared to that of wild-type mice. This means that CD169+Ly6c+F4/80low monocyte-derived macrophages in brain tumors play an important role in inhibiting tumor growth.

Analysis of the effect of CD169+Ly6c+F4/80low monocyte-derived macrophages on intratumoral immune cells

In the mouse survival experiment, it was confirmed that the mouse died faster due to the tumor in the absence of CD169+Ly6c+F4/80low monocyte-derived macrophages. Since it was confirmed that it could affect the response, it could be inferred that CD169+Ly6c+F4/80low monocyte-derived macrophages could change the immune environment in the tumor. To confirm this, changes in the immune cells in the tumor were confirmed according to the presence or absence of CD169+Ly6c+F4/80low monocyte-derived macrophages.

Experimental Method 6

In 8-week-old male CD169-DTR hetero or wild-type mice, 1000 ng of diphtheria toxin was intraperitoneally injected, and 24 hours later, 1 Х 10 of the GL261 brain tumor cell line was injected. After 4 days, the same amount of diphtheria toxin was injected intraperitoneally, and on the 10th day after injection, immune cells from the right brain hemisphere of the mouse were isolated as in Experimental Method 3. These cells were transfected with αCD45.2 (104), αF4/80 (BM8), αLy6C (AL-21), αCD11b (M1/70), αLy6G (1A8), MHCII (M5/144.15.2), CD11c (N418) antibodies was added and dyed at 4°C for 30 minutes. Before reading the cells, dead cells were stained with PI and analyzed with LSR Fortessa.

Experiment result 6

As shown in FIGS. 10, 11 and 13, the total number of immune cells (CD45.2+) and microglia (Ly6G-Ly6C-F4/80+CD45.2low), monocytes (Ly6G-F4/80) present in the tumor tissue -MHCII-Ly6C+CD11b+) and the ratio in immune cells did not show a significant difference, but as shown in FIG. 11, the number and ratio of neutrophils (Ly6G+) increased. As shown in FIG. 14, the number and ratio of monocyte-derived macrophages (Ly6G-Ly6C+F4/80+) infiltrating the tumor tissue were not statistically significant, but showed a decreasing trend, and as shown in FIG. 15, dendritic cells (Ly6G-F4/80) -MHCII+CD11c+) decreased in number and proportion. Since the changes in the immune cells in the spleen and the changes in the immune cells in the brain tumor were similar, it could be inferred that the immune cells introduced into the brain tumor were derived from the spleen and flowed into the brain through the blood. This indicates that CD169+Ly6c+F4/80low monocyte-derived macrophages not only directly affect immune cells, but also affect the permeability of immune cells in the blood vessel-brain barrier of the brain.

Analysis of the effects of CD169+Ly6c+F4/80low monocyte-derived macrophages on T-cell infiltration in glioblastoma, activity and function

According to the previous experimental results, it was confirmed that the survival period of the mice in which CD169+Ly6c+F4/80low monocyte-derived macrophages were removed was shortened in glioblastoma. This means that the anticancer immune response is suppressed. To confirm this, the infiltration of T cells, which are immune cells that play an important role in the anticancer immune response, in glioblastoma and the function of T cells present in the tumor were analyzed.

Experimental method 7

As in Experimental Method 5, mice were injected with diphtheria toxin and a tumor cell line, and then immune cells in brain tumors were obtained. The isolated immune cells were cultured for 4 hours in RPMI media containing 50 ng/ml of PMA, 1 μg/ml of inomycin, and a Golgi plug (BD Bioscience) diluted 1/1500, followed by αCD45.2 (104), CD3e (17A2), CD4 (GK1.5), CD8a (53-6.7), IFN-γ (XMG1.2), Foxp3 (FJK16s), CD44 (IM7) antibodies and Fixable viability stain 450 (BD bioscience) It was then analyzed by flow cytometry.

Experiment result 7

16, the number of total immune cells (CD45.2+) present in the tumor tissue did not show a significant difference between the groups. As shown in Figure 17, the number of CD4 T cells (CD3+CD4+) and CD8 T cells (CD3+CD8+) infiltrating the tumor tissue did not differ between the two groups in the mice injected with only DPBS, but all increased in the mice injected with the tumor. did. The number of CD8 T cells did not differ between wild-type and CD169-DTR mice, but the number of CD4 T cells decreased in CD169-DTR mice. As shown in FIG. 18 , there was little difference in the ratio of activated cells (CD4+CD44+) and cells expressing IFN-γ (CD4+IFN-γ+) among CD4 T cells in CD4 T cells, but the number was found in CD169-DTR mice. decreased. As shown in FIG. 19 , both the ratio and the number of CD4 T cells expressing activated cells (CD8+CD44+) and IFN-γ (CD8+IFN-γ+) among CD8 T cells were decreased in CD169-DTR mice. As shown in FIG. 20, in the case of regulatory T cells (CD4+Foxp3+), both the ratio and the number of cells in the CD4 T cells were decreased.

According to these results, in the absence of CD169+Ly6c+F4/80low monocyte-derived macrophages, the infiltration, function and activity of CD4 T cells and CD8 T cells, which are important for the anticancer immune response, in the tumor are reduced, and regulatory T cells that suppress the anticancer immune response. It can be seen that the penetration of

Analysis of the effect of CD169+Ly6c+F4/80low monocyte-derived macrophages on the immune environment in glioblastoma

In mice with CD169+Ly6c+F4/80low monocyte-derived macrophages removed, the tumor infiltration, activity, and function of T cells decreased, suggesting that CD169+ macrophages affect the immune environment in glioblastoma. As a result of single-cell transcriptome analysis, CD169+Ly6c+F4/80low monocyte-derived macrophages mainly expressed CXCL9 and CXCL10. To confirm this, the expression of chemokines and cytokines regulating the movement, function and activity of T cells was analyzed.

Experimental method 8

As in Experimental Method 5, mice were injected with diphtheria toxin and a tumor cell line, and 10 days later, the right hemisphere tissue of the brain was isolated. Put the isolated tissue in a strainer with 1 ml RPMI media, grind it finely, centrifuge it for 5 minutes at 500 g, and extract RNA from the submerged tissue using RNeasy Plus Mini Kit (Qiagen) and ReverTra Ace qPCR RT Master Mix ( Toyobo) was used to synthesize cDNA.

The expression of chemokines and cytokines was analyzed using the synthesized cDNA SYBR Green Realitime PCR Master Mix (Toyobo) and primers (Table 1).

Experiment result 8

As shown in FIG. 21 , CXCL9 among chemokines important for infiltration of T cells into the tumor did not differ between wild-type and CD169-DTR mice, but CXCL10 was increased, and CCL22, which is important for influx of regulatory T cells, was increased in CD169-DTR mice. From these results, it could be inferred that the change in chemokine expression in the tumor mediates the change in the influx of regulatory T cells. In the case of cytokines, the expression of IL-12p40 was decreased in CD169-DTR mice, although there was no significant difference between other cytokines as shown in FIG. 22 . IL-12 is a cytokine mainly expressed in innate immune cells, such as dendritic cells, and plays an important role in the activation of T cells. These results indicate that the function and influx of various immune cells in the tumor are regulated by CD169+Ly6c+F4/80low monocyte-derived macrophages, together with the previous experimental results in which the influx of dendritic cells into the tumor was reduced in CD169-DTR mice. do. In addition, in order to confirm the degree of hypoxia, which is known to affect the activity of immune cells in the tumor, as shown in FIG. 23 , the expression of VEGF and HIF-1α increased in hypoxia was confirmed. As a result, there was no difference in the expression of VEGF, but HIF-1α expression was increased in CD169-DTR mice. This means that CD169+Ly6c+F4/80low monocyte-derived macrophages can reduce intratumoral hypoxia.

As the specific parts of the present invention have been described in detail above, for those of ordinary skill in the art, these specific techniques are only preferred embodiments, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

<110> Korea Advanced Institute of science and technology <120> Cell surface antigen of Macrophage and the use thereof <130> PDPB194149 <160> 5 <170> KoPatentIn 3.0 <210> 1 <211> 1695 <212> PRT <213> Homo sapiens <400> 1 Met Cys Val Leu Phe Ser Leu Leu Leu Leu Ala Ser Val Phe Ser Leu 1 5 10 15 Gly Gln Thr Thr Trp Gly Val Ser Ser Pro Lys Asn Val Gln Gly Leu 20 25 30 Ser Gly Ser Cys Leu Leu Ile Pro Cys Ile Phe Ser Tyr Pro Ala Asp 35 40 45 Val Pro Val Ser Asn Gly Ile Thr Ala Ile Trp Tyr Tyr Asp Tyr Ser 50 55 60 Gly Lys Arg Gln Val Val Ile His Ser Gly Asp Pro Lys Leu Val Asp 65 70 75 80 Lys Arg Phe Arg Gly Arg Ala Glu Leu Met Gly Asn Met Asp His Lys 85 90 95 Val Cys Asn Leu Leu Leu Lys Asp Leu Lys Pro Glu Asp Ser Gly Thr 100 105 110 Tyr Asn Phe Arg Phe Glu Ile Ser Asp Ser Asn Arg Trp Leu Asp Val 115 120 125 Lys Gly Thr Thr Val Thr Val Thr Thr Asp Pro Ser Pro Pro Thr Ile 130 135 140 Thr Ile Pro Glu Glu Leu Arg Glu Gly Met Glu Arg Asn Phe Asn Cys 145 150 155 160 Ser Thr Pro Tyr Leu Cys Leu Gln Glu Lys Gln Val Ser Leu Gln Trp 165 170 175 Arg Gly Gln Asp Pro Thr His Ser Val Thr Ser Ser Phe Gln Ser Leu 180 185 190 Glu Pro Thr Gly Val Tyr His Gln Thr Thr Leu His Met Ala Leu Ser 195 200 205 Trp Gln Asp His Gly Arg Thr Leu Leu Cys Gln Phe Ser Leu Gly Ala 210 215 220 His Ser Ser Arg Lys Glu Val Tyr Leu Gln Val Pro His Ala Pro Lys 225 230 235 240 Gly Val Glu Ile Leu Leu Ser Ser Ser Gly Arg Asn Ile Leu Pro Gly 245 250 255 Asp Pro Val Thr Leu Thr Cys Arg Val Asn Ser Ser Tyr Pro Ala Val 260 265 270 Ser Ala Val Gln Trp Ala Arg Asp Gly Val Asn Leu Gly Val Thr Gly 275 280 285 His Val Leu Arg Leu Phe Ser Ala Ala Trp Asn Asp Ser Gly Ala Tyr 290 295 300 Thr Cys Gln Ala Thr Asn Asp Met Gly Ser Leu Val Ser Pro Leu 305 310 315 320 Ser Leu His Val Phe Met Ala Glu Val Lys Met Asn Pro Ala Gly Pro 325 330 335 Val Leu Glu Asn Glu Thr Val Thr Leu Leu Cys Ser Thr Pro Lys Glu 340 345 350 Ala Pro Gln Glu Leu Arg Tyr Ser Trp Tyr Lys Asn His Ile Leu Leu 355 360 365 Glu Asp Ala His Ala Ser Thr Leu His Leu Pro Ala Val Thr Arg Ala 370 375 380 Asp Thr Gly Phe Tyr Phe Cys Glu Val Gln Asn Ala Gln Gly Ser Glu 385 390 395 400 Arg Ser Ser Pro Leu Ser Val Val Val Arg Tyr Pro Leu Thr Pro 405 410 415 Asp Leu Thr Thr Phe Leu Glu Thr Gln Ala Gly Leu Val Gly Ile Leu 420 425 430 His Cys Ser Val Val Ser Glu Pro Leu Ala Thr Val Val Leu Ser His 435 440 445 Gly Gly Leu Thr Leu Ala Ser Asn Ser Gly Glu Asn Asp Phe Asn Pro 450 455 460 Arg Phe Arg Ile Ser Ser Ala Pro Asn Ser Leu Arg Leu Glu Ile Arg 465 470 475 480 Asp Leu Gln Pro Ala Asp Ser Gly Glu Tyr Thr Cys Leu Ala Val Asn 485 490 495 Ser Leu Gly Asn Ser Thr Ser Ser Leu Asp Phe Tyr Ala Asn Val Ala 500 505 510 Arg Leu Leu Ile Asn Pro Ser Ala Glu Val Val Glu Gly Gln Ala Val 515 520 525 Thr Leu Ser Cys Arg Ser Gly Leu Ser Pro Ala Pro Asp Thr Arg Phe 530 535 540 Ser Trp Tyr Leu Asn Gly Ala Leu Leu Leu Glu Gly Ser Ser Ser Ser Ser 545 550 555 560 Leu Leu Leu Pro Ala Ala Ser Ser Thr Asp Ala Gly Ser Tyr Tyr Cys 565 570 575 Arg Thr Gln Ala Gly Pro Asn Thr Ser Gly Pro Ser Leu Pro Thr Val 580 585 590 Leu Thr Val Phe Tyr Pro Pro Arg Lys Pro Thr Phe Thr Ala Arg Leu 595 600 605 Asp Leu Asp Thr Ser Gly Val Gly Asp Gly Arg Arg Gly Ile Leu Leu 610 615 620 Cys His Val Asp Ser Asp Pro Pro Ala Gln Leu Arg Leu Leu His Lys 625 630 635 640 Gly His Val Val Ala Thr Ser Leu Pro Ser Arg Cys Gly Ser Cys Ser 645 650 655 Gln Arg Thr Lys Val Ser Arg Thr Ser Asn Ser Leu His Val Glu Ile 660 665 670 Gln Lys Pro Val Leu Glu Asp Glu Gly Val Tyr Leu Cys Glu Ala Ser 675 680 685 Asn Thr Leu Gly Asn Ser Ser Ala Ala Ala Ser Phe Asn Ala Lys Ala 690 695 700 Thr Val Leu Val Ile Thr Pro Ser Asn Thr Leu Arg Glu Gly Thr Glu 705 710 715 720 Ala Asn Leu Thr Cys Asn Val Asn Gln Glu Val Ala Val Ser Pro Ala 725 730 735 Asn Phe Ser Trp Phe Arg Asn Gly Val Leu Trp Thr Gln Gly Ser Leu 740 745 750 Glu Thr Val Arg Leu Gln Pro Val Ala Arg Thr Asp Ala Ala Val Tyr 755 760 765 Ala Cys Arg Leu Leu Thr Glu Asp Gly Ala Gln Leu Ser Ala Pro Val 770 775 780 Val Leu Ser Val Leu Tyr Ala Pro Asp Pro Pro Lys Leu Ser Ala Leu 785 790 795 800 Leu Asp Val Gly Gln Gly His Met Ala Val Phe Ile Cys Thr Val Asp 805 810 815 Ser Tyr Pro Leu Ala His Leu Ser Leu Phe Arg Gly Asp His Leu Leu 820 825 830 Ala Thr Asn Leu Glu Pro Gln Arg Pro Ser His Gly Arg Ile Gln Ala 835 840 845 Lys Ala Thr Ala Asn Ser Leu Gln Leu Glu Val Arg Glu Leu Gly Leu 850 855 860 Val Asp Ser Gly Asn Tyr His Cys Glu Ala Thr Asn Ile Leu Gly Ser 865 870 875 880 Ala Asn Ser Ser Leu Phe Phe Gln Val Arg Gly Ala Trp Val Gln Val 885 890 895 Ser Pro Ser Pro Glu Leu Arg Glu Gly Gln Ala Val Val Leu Ser Cys 900 905 910 Gln Val Pro Thr Gly Val Ser Glu Gly Thr Ser Tyr Ser Trp Tyr Gln 915 920 925 Asp Gly Arg Pro Leu Gln Glu Ser Thr Ser Ser Thr Leu Arg Ile Ala 930 935 940 Ala Ile Ser Leu Arg Gln Ala Gly Ala Tyr His Cys Gln Ala Gln Ala 945 950 955 960 Pro Asp Thr Ala Ile Ala Ser Leu Ala Ala Pro Val Ser Leu His Val 965 970 975 Ser Tyr Thr Pro Arg His Val Thr Leu Ser Ala Leu Leu Ser Thr Asp 980 985 990 Pro Glu Arg Leu Gly His Leu Val Cys Ser Val Gln Ser Asp Pro Pro 995 1000 1005 Ala Gln Leu Gln Leu Phe His Arg Asn Arg Leu Val Ala Ser Thr Leu 1010 1015 1020 Gln Gly Ala Asp Glu Leu Ala Gly Ser Asn Pro Arg Leu His Val Thr 1025 1030 1035 1040 Val Leu Pro Asn Glu Leu Arg Leu Gln Ile His Phe Pro Glu Leu Glu 1045 1050 1055 Asp Asp Gly Thr Tyr Thr Cys Glu Ala Ser Asn Thr Leu Gly Gly Gln Ala 1060 1065 1070 Ser Ala Ala Ala Asp Phe Asp Ala Gln Ala Val Arg Val Thr Val Trp 1075 1080 1085 Pro Asn Ala Thr Val Gln Glu Gly Gin Gln Val Asn Leu Thr Cys Leu 1090 1095 1100 Val Trp Ser Thr His Gln Asp Ser Leu Ser Tyr Thr Trp Tyr Lys Gly 1105 1110 1115 1120 Gly Gln Gln Leu Leu Gly Ala Arg Ser Ile Thr Leu Pro Ser Val Lys 1125 1130 1135 Val Leu Asp Ala Thr Ser Tyr Arg Cys Gly Val Gly Leu Pro Gly His 1140 1145 1150 Ala Pro His Leu Ser Arg Pro Val Thr Leu Asp Val Leu His Ala Pro 1155 1160 1165 Arg Asn Leu Arg Leu Thr Tyr Leu Leu Glu Thr Gln Gly Arg Gln Leu 1170 1175 1180 Ala Leu Val Leu Cys Thr Val Asp Ser Arg Pro Pro Ala Gln Leu Thr 1185 1190 1195 1200 Leu Ser His Gly Asp Gln Leu Val Ala Ser Ser Thr Glu Ala Ser Val 1205 1210 1215 Pro Asn Thr Leu Arg Leu Glu Leu Gln Asp Pro Arg Pro Ser Asn Glu 1220 1225 1230 Gly Leu Tyr Ser Cys Ser Ala His Ser Pro Leu Gly Lys Ala Asn Thr 1235 1240 1245 Ser Leu Glu Leu Leu Leu Glu Gly Val Arg Val Lys Met Asn Pro Ser 1250 1255 1260 Gly Ser Val Pro Glu Gly Glu Pro Val Thr Val Thr Cys Glu Asp Pro 1265 1270 1275 1280 Ala Ala Leu Ser Ser Ala Leu Tyr Ala Trp Phe His Asn Gly His Trp 1285 1290 1295 Leu Gln Glu Gly Pro Ala Ser Ser Leu Gln Phe Leu Val Thr Thr Arg 1300 1305 1310 Ala His Ala Gly Ala Tyr Phe Cys Gln Val His Asp Thr Gln Gly Thr 1315 1320 1325 Arg Ser Ser Arg Pro Ala Ser Leu Gln Ile Leu Tyr Ala Pro Arg Asp 1330 1335 1340 Ala Val Leu Ser Ser Phe Arg Asp Ser Arg Thr Arg Leu Met Val Val 1345 1350 1355 1360 Ile Gln Cys Thr Val Asp Ser Glu Pro Ala Glu Met Val Leu Ser 1365 1370 1375 His Asn Gly Lys Val Leu Ala Ala Ser His Glu Arg His Ser Ser Ala 1380 1385 1390 Ser Gly Ile Gly His Ile Gln Val Ala Arg Asn Ala Leu Arg L eu Gln 1395 1400 1405 Val Gln Asp Val Thr Leu Gly Asp Gly Asn Thr Tyr Val Cys Thr Ala 1410 1415 1420 Gln Asn Thr Leu Gly Ser Ile Ser Thr Thr Gln Arg Leu Leu Thr Glu 1425 1430 1435 1440 Thr Asp Ile Arg Val Thr Ala Glu Pro Gly Leu Asp Val Pro Glu Gly 1445 1450 1455 Thr Ala Leu Asn Leu Ser Cys Leu Leu Pro Gly Gly Ser Gly Pro Thr 1460 1465 1470 Gly Asn Ser Ser Phe Thr Trp Phe Trp Asn Arg His Arg Leu His Ser 1475 1480 1485 Ala Pro Val Pro Thr Leu Ser Phe Thr Pro Val Val Arg Ala Gln Ala 1490 1495 1500 Gly Leu Tyr His Cys Arg Ala Asp Leu Pro Thr Gly Ala Thr Thr Ser 1505 1510 1515 1520 Ala Pro Val Met Leu Arg Val Leu Tyr Pro Pro Lys Thr Pro Thr Leu 1525 1530 1535 Ile Val Phe Val Glu Pro Gln Gly Gly His Gln Gly Ile Leu Asp Cys 1540 1545 1550 Arg Val Asp Ser Glu Pro Leu Ala Ile Leu Thr Leu His Arg Gly Ser 1555 1560 1565 Gln Leu Val Ala Ser Asn Gln Leu His Asp Ala Pro Thr Lys Pro His 1570 1575 1580 Ile Arg Val Thr Ala Pro Pro Asn Ala Leu Arg Val Asp Ile Glu Glu 1585 1590 1595 1600 Leu Gly Pro Ser Asn Gln Gly Glu Tyr Val Cys Thr Ala Ser Asn Thr 1605 1610 1615 Leu Gly Ser Ala Ser Ala Ser Ala Tyr Phe Gly Thr Arg Ala Leu His 1620 1625 1630 Gln Leu Gln Leu Phe Gln Arg Leu Leu Trp Val Leu Gly Phe Leu Ala 1635 1640 1645 Gly Phe Leu Cys Leu Leu Leu Gly Leu Val Ala Tyr His Thr Trp Arg 1650 1655 1660 Lys Lys Ser Ser Thr Lys Leu Asn Glu Asp Glu Asn Ser Ala Glu Met 1665 1670 1675 1680 Ala Thr Lys Lys Asn Thr Ile Gln Glu Glu Val Val Ala Ala Leu 1685 1690 1695 <210> 2 <211> 1293 <212> PRT <213> Homo sapiens <400> 2 Met Thr Met Gly Leu Trp Leu Lys Leu Leu Ala Phe Gly Phe Ala Leu 1 5 10 15 Leu Asp Thr Glu Val Phe Val Thr Gly Gln Thr Pro Thr Pro Ser Asp 20 25 30 Glu Leu Ser Thr Thr Glu Asn Ala Leu Leu Leu Pro Gln Ser Asp Pro 35 40 45 Leu Pro Ala Arg Thr Thr Glu Ser Thr Pro Pro Ser Ile Ser Glu Arg 50 55 60 Gly Asn Gly Ser Ser Glu Thr Thr Tyr His Pro Gly Val Leu Ser Thr 65 70 75 80 Leu Leu Pro His Leu Ser Pro Gln Pro Asp Ser Gln Thr Pro Ser Ala 85 90 95 Gly Gly Ala Asp Thr Gln Thr Phe Ser Ser Gln Ala Asp Asn Pro Thr 100 105 110 Leu Thr Pro Ala Pro Gly Gly Gly Thr Asp Pro Pro Gly Val Pro Gly 115 120 125 Glu Arg Thr Val Pro Gly Thr Ile Pro Ala Asp Thr Ala Phe Pro Val 130 135 140 Asp Thr Pro Ser Leu Ala Arg Asn Ser Ser Ala Ala Ser Pro Thr His 145 150 155 160 Thr Ser Asn Val Ser Thr Thr Asp Ile Ser Ser Gly Ala Ser Leu Thr 165 170 175 Thr Leu Thr Pro Ser Thr Leu Gly Leu Ala Ser Thr Asp Pro Pro Ser 180 185 190 Thr Thr Ile Ala Thr Thr Thr Lys Gln Thr Cys Ala Ala Met Phe Gly 195 200 205 Asn Ile Thr Val Asn Tyr Thr Tyr Glu Ser Ser Asn Gln Thr Phe Lys 210 215 220 Ala Asp Leu Lys Asp Val Gln Asn Ala Lys Cys Gly Asn Glu Asp Cys 225 230 235 240 Glu Asn Val Leu Asn Asn Leu Glu Glu Cys Ser Gln Ile Lys Asn Ile 245 250 255 Ser Val Ser Asn Asp Ser Cys Ala Pro Ala Thr Thr Ile Asp Leu Tyr 260 265 270 Val Pro Pro Gly Thr Asp Lys Phe Ser Leu His Asp Cys Thr Pro Lys 275 280 285 Glu Lys Ala Asn Thr Ser Ile Cys Leu Glu Trp Lys Thr Lys Asn Leu 290 295 300 Asp Phe Arg Lys Cys Asn Ser Asp Asn Ile Ser Tyr Val Leu His Cys 305 310 315 320 Glu Pro Glu Asn Asn Thr Lys Cys Ile Arg Arg Asn Thr Phe Ile Pro 325 330 335 Glu Arg Cys Gln Leu Asp Asn Leu Arg Ala Gln Thr Asn Tyr Thr Cys 340 345 350 Val Ala Glu Ile Leu Tyr Arg Gly Val Lys Leu Val Lys Asn Val Ile 355 360 365 Asn Val Gln Thr Asp Leu Gly Ile Pro Glu Thr Pro Lys Pro Ser Cys 370 375 380 Gly Asp Pro Ala Ala Arg Lys Thr Leu Val Ser Trp Pro Glu Pro Val 385 390 395 400 Ser Lys Pro Glu Ser Ala Ser Lys Pro His Gly Tyr Val Leu Cys Tyr 405 410 415 Lys Asn Asn Ser Glu Lys Cys Lys Ser Leu Pro Asn Asn Val Thr Ser 420 425 430 Phe Glu Val Glu Ser Leu Lys Pro Tyr Lys Tyr Tyr Glu Val Ser Leu 435 440 445 Leu Ala Tyr Val Asn Gly Lys Ile Gln Arg Asn Gly Thr Ala Glu Lys 450 455 460 Cys Asn Phe His Thr Lys Ala Asp Arg Pro Asp Lys Val Asn Gly Met 465 470 475 480 Lys Thr Ser Arg Pro Thr Asp Asn Ser Ile Asn Val Thr Cys Gly Pro 485 490 495 Pro Tyr Glu Thr Asn Gly Pro Lys Thr Phe Tyr Ile Leu Val Val Arg 500 505 510 Ser Gly Gly Ser Phe Val Thr Lys Tyr Asn Lys Thr Asn Cys Gln Phe 515 520 525 Tyr Val Asp Asn Leu Tyr Tyr Ser Thr Asp Tyr Glu Phe Leu Val Ser 530 535 540 Phe His Asn Gly Val Tyr Glu Gly Asp Ser Val Ile Arg Asn Glu Ser 545 550 555 560 Thr Asn Phe Asn Ala Lys Ala Leu Ile Ile Phe Leu Val Phe Leu Ile 565 570 575 Ile Val Thr Ser Ile Ala Leu Leu Val Val Leu Tyr Lys Ile Tyr Asp 580 585 590 Leu Arg Lys Lys Arg Ser Ser Asn Leu Asp Glu Gln Gln Glu Leu Val 595 600 605 Glu Arg Asp Asp Glu Lys Gln Leu Met Asp Val Glu Pro Ile His Ser 610 615 620 Asp Ile Leu Leu Glu Thr Tyr Lys Arg Lys Ile Ala Asp Glu Gly Arg 625 630 635 640 Leu Phe Leu Ala Glu Phe Gln Ser Ile Pro Arg Val Phe Ser Lys Phe 645 650 655 Pro Ile Lys Asp Ala Arg Lys Pro His Asn Gln Asn Lys Asn Arg Tyr 660 665 670 Val Asp Ile Leu Pro Tyr Asp Tyr Asn Arg Val Glu Leu Ser Glu Ile 675 680 685 Asn Gly Asp Ala Gly Ser Thr Tyr Ile Asn Ala Ser Tyr Ile Asp Gly 690 695 700 Phe Lys Glu Pro Arg Lys Tyr Ile Ala Ala Gln Gly Pro Arg Asp Glu 705 710 715 720 Thr Val Asp Asp Phe Trp Arg Met Ile Trp Glu Gln Lys Ala Thr Val 725 730 735 Ile Val Met Val Thr Arg Cys Glu Glu Gly Asn Arg Asn Lys Cys Ala 740 745 750 Glu Tyr Trp Pro Ser Met Glu Glu Gly Thr Arg Ala Phe Lys Asp Ile 755 760 765 Val Val Thr Ile Asn Asp His Lys Arg Cys Pro Asp Tyr Ile Ile Gln 770 775 780 Lys Leu Asn Val Ala His Lys Lys Glu Lys Ala Thr Gly Arg Glu Val 785 790 795 800 Thr His Ile Gln Phe Thr Ser Trp Pro Asp His Gly Val Pro Glu Asp 805 810 815 Pro His Leu Leu Leu Leu Lys Leu Arg Arg Arg Val Asn Ala Phe Ser Asn 820 825 830 Phe Phe Ser Gly Pro Ile Val Val His Cys Ser Ala Gly Val Gly Arg 835 840 845 Thr Gly Thr Tyr Ile Gly Ile Asp Ala Met Leu Glu Gly Leu Glu Ala 850 855 860 Glu Gly Lys Val Asp Val Tyr Gly Tyr Val Val Lys Leu Arg Arg Gln 865 870 875 880 Arg Cys Leu Met Val Gln Val Glu Ala Gln Tyr Ile Leu Ile His Gln 885 890 895 Ala Leu Val Glu Tyr Asn Gln Phe Gly Glu Thr Glu Val Asn Leu Ser 900 905 910 Glu Leu His Ser Cys Leu His Asn Met Lys Lys Arg Asp Pro Ser 915 920 925 Asp Pro Ser Pro Leu Glu Ala Glu Tyr Gln Arg Leu Pro Ser Tyr Arg 930 935 940 Ser Trp Arg Thr Gln His Ile Gly Asn Gln Glu Glu Asn Lys Lys Lys 945 950 955 960 Asn Arg Asn Ser Asn Val Val Pro Tyr Asp Phe Asn Arg Val Pro Leu 965 970 975 Lys His Glu Leu Glu Met Ser Lys Glu Ser Glu Pro Glu Ser Asp Glu 980 985 990 Ser Ser Asp Asp Asp Ser Asp Ser Glu Glu Thr Ser Lys Tyr Ile Asn 995 1000 1005 Ala Ser Phe Val Met Ser Tyr Trp Lys Pro Glu Met Met Ile Ala Ala 1010 1015 1020 Gln Gly Pro Leu Lys Glu Thr Ile Gly Asp Phe Trp Gln Met Ile Phe 1025 1030 1035 1040 Gln Arg Lys Val Lys Val Ile Val Met Leu Thr Glu Leu Val Asn Gly 1045 1050 1055 Asp Gln Glu Val Cys Ala Gln Tyr Trp Gly Glu Gly Lys Gln Thr Tyr 1060 1065 1070 Gly Asp Met Glu Val Glu Met Lys Asp Thr Asn Arg Ala Ser Ala Tyr 1075 1080 1085 Thr Leu Arg Thr Phe Glu Leu Arg His Ser Lys Arg Lys Glu Pro Arg 1090 1095 1100 Thr Val Tyr Gln Tyr Gln Cys Thr Thr Trp Lys Gly Glu Glu Leu Pro 1105 1110 1115 1120 Ala Glu Pro Lys Asp Leu Val Ser Met Ile Gln Asp Leu Lys Gln Lys 1125 1130 1135 Leu Pro Lys Ala Ser Pro Glu Gly Met Lys Tyr His Lys His Ala Ser 1140 1145 1150 Ile Leu Val His Cys Arg Asp Gly Ser Gln Gln Thr Gly Leu Phe Cys 1155 1160 1165 Ala Leu Phe Asn Leu Leu Glu Ser Ala Glu Thr Glu Asp Val Val Asp 1170 1175 1180 Val Phe Gln Val Val Lys Ser Leu Arg Lys Ala Arg Pro Gly Val Val 1185 1190 1195 1200 Cys Ser Tyr Glu Gln Tyr Gln Phe Leu Tyr Asp Ile Ile Ala Ser Ile 1205 1210 1215 Tyr Pro Ala Gln Asn Gly Gln Val Lys Lys Thr Asn Ser Gln Asp Lys 1220 1225 1230 Ile Glu Phe His Asn Glu Val Asp Gly Gly Lys Gln Asp Ala Asn Cys 1235 1240 1245 Val Arg Pro Asp Gly Pro Leu Asn Lys Ala Gln Glu Asp Ser Arg Gly 1250 1255 1260 Val Gly Thr Pro Glu Pro Thr Asn Ser Ala Glu Glu Pro Glu His Ala 1265 1270 1275 1280 Ala Asn Gly Ser Ala Ser Pro Ala Pro Thr Gln Ser Ser 1285 1290 <210> 3 <211> 931 <212> PRT <213> Homo sapiens <400> 3 Met Trp Gly Phe Trp Leu Leu Leu Phe Trp Gly Phe Ser Gly Met Tyr 1 5 10 15 Arg Trp Gly Met Thr Thr Leu Pro Thr Leu Gly Gln Thr Leu Gly Gly 20 25 30 Val Asn Glu Cys Gln Asp Thr Thr Thr Cys P ro Ala Tyr Ala Thr Cys 35 40 45 Thr Asp Thr Thr Asp Ser Tyr Tyr Cys Thr Cys Lys Arg Gly Phe Leu 50 55 60 Ser Ser Asn Gly Gln Thr Asn Phe Gln Gly Pro Gly Val Glu Cys Gln 65 70 75 80 Asp Val Asn Glu Cys Leu Gln Ser Asp Ser Pro Cys Gly Pro Asn Ser 85 90 95 Val Cys Thr Asn Ile Leu Gly Arg Ala Lys Cys Ser Cys Leu Arg Gly 100 105 110 Phe Ser Ser Ser Thr Gly Lys Asp Trp Ile Leu Gly Ser Leu Asp Asn 115 120 125 Phe Leu Cys Ala Asp Val Asp Glu Cys Leu Thr Ile Gly Ile Cys Pro 130 135 140 Lys Tyr Ser Asn Cys Ser Asn Ser Val Gly Ser Tyr Ser Cys Thr Cys 145 150 155 160 Gln Pro Gly Phe Val Leu Asn Gly Ser Ile Cys Glu Asp Glu Asp Glu 165 170 175 Cys Val Thr Arg Asp Val Cys Pro Glu His Ala Thr Cys His Asn Thr 180 185 190 Leu Gly Ser Tyr Tyr Cys Thr Cys Asn Ser Gly Leu Glu Ser Ser Gly 195 200 205 Gly Gly Pro Met Phe Gln Gly Leu Asp Glu Ser Cys Glu Asp Val Asp 210 215 220 Glu Cys Ser Arg Asn Ser Thr Leu Cys Gly Pro Thr Phe Ile Cys Ile 225 230 235 240 Asn Thr Leu Gly Ser Tyr Ser Cys Ser Cys Pro Ala Gly Phe Ser Leu 245 250 255 Pro Thr Phe Gln Ile Leu Gly His Pro Ala Asp Gly Asn Cys Thr Asp 260 265 270 Ile Asp Glu Cys Asp Asp Thr Cys Pro Leu Asn Ser Ser Cys Thr Asn 275 280 285 Thr Ile Gly Ser Tyr Phe Cys Thr Cys His Pro Gly Phe Ala Ser Ser 290 295 300 Asn Gly Gln Leu Asn Phe Lys Asp Leu Glu Val Thr Cys Glu Asp Ile 305 310 315 320 Asp Glu Cys Thr Gln Asp Pro Leu Gln Cys Gly Leu Asn Ser Val Cys 325 330 335 Thr Asn Val Pro Gly Ser Tyr Ile Cys Gly Cys Leu Pro Asp Phe Gln 34 0 345 350 Met Asp Pro Glu Gly Ser Gln Gly Tyr Gly Asn Phe Asn Cys Lys Arg 355 360 365 Ile Leu Phe Lys Cys Lys Glu Asp Leu Ile Leu Gln Ser Glu Gln Ile 370 375 380 Gln Gln Cys Gln Ala Val Gln Gly Arg Asp Leu Gly Tyr Ala Ser Phe 385 390 395 400 Cys Thr Leu Val Asn Ala Thr Phe Thr Ile Leu Asp Asn Thr Cys Glu 405 410 415 Asn Lys Ser Ala Pro Val Ser Leu Gln Ser Ala Ala Thr Ser Val Ser 420 425 430 Leu Val Leu Glu Gln Ala Thr Thr Trp Phe Glu Leu Ser Lys Glu Glu 435 440 445 Thr Ser Thr Leu Gly Thr Ile Leu Leu Glu Thr Val Glu Ser Thr Met 450 455 460 Leu Ala Ala Leu Leu Ile Pro Ser Gly Asn Ala Ser Gln Met Ile Gln 465 470 475 480 Thr Glu Tyr Leu Asp Ile Glu Ser Lys Val Ile Asn Glu Gl u Cys Lys 485 490 495 Glu Asn Glu Ser Ile Asn Leu Ala Ala Arg Gly Asp Lys Met Asn Val 500 505 510 Gly Cys Phe Ile Lys Glu Ser Val Ser Thr Gly Ala Pro Gly Val 515 520 525 Ala Phe Val Ser Phe Ala His Met Glu Ser Val Leu Asn Glu Arg Phe 530 535 540 Phe Glu Asp Gly Gln Ser Phe Arg Lys Leu Arg Met Asn Ser Arg Val 545 550 555 560 Val Gly Gly Thr Val Thr Gly Glu Lys Lys Glu Asp Phe Ser Lys Pro 565 570 575 Ile Ile Tyr Thr Leu Gln His Ile Gln Pro Lys Gln Lys Ser Glu Arg 580 585 590 Pro Ile Cys Val Ser Trp Asn Thr Asp Val Glu Asp Gly Arg Trp Thr 595 600 605 Pro Ser Gly Cys Glu Ile Val Glu Ala Ser Glu Thr His Thr Val Cys 610 615 620 Ser Cys Asn Arg Met Ala Asn Leu Ala Ile Ile Met Ala Ser Gly Gl u 625 630 635 640 Leu Thr Met Glu Phe Ser Leu Tyr Ile Ile Ser His Val Gly Thr Val 645 650 655 Ile Ser Leu Val Cys Leu Ala Leu Ala Ile Ala Thr Phe Leu Leu Cys 660 665 670 Arg Ala Val Gln Asn His Asn Thr Tyr Met His Leu His Leu Cys Val 675 680 685 Cys Leu Phe Leu Ala Lys Ile Leu Phe Leu Thr Gly Ile Asp Lys Thr 690 695 700 Asp Asn Gln Thr Ala Cys Ala Ile Ile Ala Gly Phe Leu His Tyr Leu 705 710 715 720 Phe Leu Ala Cys Phe Phe Trp Met Leu Val Glu Ala Val Met Leu Phe 725 730 735 Leu Met Val Arg Asn Leu Lys Val Val Val Asn Tyr Phe Ser Ser Arg Asn 740 745 750 Ile Lys Met Leu His Leu Cys Ala Phe Gly Tyr Gly Leu Pro Val Leu 755 760 765 Val Val Ile Ile Ser Ala Ser Val Gln Pro Arg Gly Ty r Gly Met His 770 775 780 Asn Arg Cys Trp Leu Asn Thr Glu Thr Gly Phe Ile Trp Ser Phe Leu 785 790 795 800 Gly Pro Val Cys Met Ile Ile Thr Ile Asn Ser Val Leu Leu Ala Trp 805 810 815 Thr Leu Trp Val Leu Arg Gln Lys Leu Cys Ser Val Ser Ser Glu Val 820 825 830 Ser Lys Leu Lys Asp Thr Arg Leu Leu Thr Phe Lys Ala Ile Ala Gln 835 840 845 Ile Phe Ile Leu Gly Cys Ser Trp Val Leu Gly Ile Phe Gln Ile Gly 850 855 860 Pro Leu Ala Ser Ile Met Ala Tyr Leu Phe Thr Ile Ile Asn Ser Leu 865 870 875 880 Gln Gly Ala Phe Ile Phe Leu Ile His Cys Leu Leu Asn Arg Gln Val 885 890 895 Arg Asp Glu Tyr Lys Lys Leu Leu Thr Arg Lys Thr Asp Leu Ser Ser 900 905 910 His Ser Gln Thr Ser Gly Ile Leu Leu Se r Ser Met Pro Ser Thr Ser 915 920 925 Lys Met Gly 930 <210> 4 <211> 131 <212> PRT <213> Mus musculus <400> 4 Met Asp Ser Thr His Ala Thr Lys Ser Cys Leu Leu Ile Leu Leu Val 1 5 10 15 Ala Leu Leu Cys Ala Gly Arg Ala Gln Gly Leu Gln Cys Tyr Glu Cys 20 25 30 Tyr Gly Val Pro Ile Glu Thr Ser Cys Pro Ala Val Thr Cys Arg Ala 35 40 45 Ser Asp Gly Phe Cys Ile Ala Gln Asn Ile Glu Leu Ile Glu Asp Ser 50 55 60 Gln Arg Arg Lys Leu Lys Thr Arg Gln Cys Leu Ser Phe Cys Pro Ala 65 70 75 80 Gly Val Pro Ile Lys Asp Pro Asn Ile Arg Glu Arg Thr Ser Cys Cys 85 90 95 Ser Glu Asp Leu Cys Asn Ala Ala Val Pro Thr Ala Gly Ser Thr Trp 100 105 110 Thr Met Ala Gly Val Leu Leu Phe Ser Leu Ser Ser Val Ile Leu Gln 115 120 125 Thr Leu Leu 130 <210> 5 <211> 375 <212> PRT <213> Homo sapiens <400> 5 Met Glu Arg Ala Ser Cys Leu Leu Leu Leu Leu Leu Leu Pro Leu Val His 1 5 10 15 Val Ser Ala Thr Thr Pro Glu Pro Cys Glu Leu Asp Asp Glu Asp Phe 20 25 30 Arg Cys Val Cys Asn Phe Ser Glu Pro Gln Pro Asp Trp Ser Glu Ala 35 40 45 Phe Gln Cys Val Ser Ala Val Glu Val Glu Ile His Ala Gly Gly Leu 50 55 60 Asn Leu Glu Pro Phe Leu Lys Arg Val Asp Ala Asp Ala Asp Pro Arg 65 70 75 80 Gln Tyr Ala Asp Thr Val Lys Ala Leu Arg Val Arg Arg Leu Thr Val 85 90 95 Gly Ala Ala Gln Val Pro Ala Gln Leu Leu Val Gly Ala Leu Arg Val 100 105 110 Leu Ala Tyr Ser Arg Leu Lys Glu Leu Thr Leu Glu Asp Leu Lys Ile 115 120 125 Thr Gly Thr Met Pro Leu Pro Leu Glu Ala Thr Gly Leu Ala Leu 130 135 140 Ser Ser Leu Arg Leu Arg Asn Val Ser Trp Ala Thr Gly Arg Ser Trp 145 150 155 160 Leu Ala Glu Leu Gln Gln Trp Leu Lys Pro Gly Leu Lys Val Leu Ser 165 170 175 Ile Ala Gln Ala His Ser Pro Ala Phe Ser Cys Glu Gln Val Arg Ala 180 185 190 Phe Pro Ala Leu Thr Ser Leu Asp Leu Ser Asp Asn Pro Gly Leu Gly 195 200 205 Glu Arg Gly Leu Met Ala Ala Leu Cys Pro His Lys Phe Pro Ala Ile 210 215 220 Gln Asn Leu Ala Leu Arg Asn Thr Gly Met Glu Thr Pro Thr Gly Val 225 230 235 240 Cys Ala Ala Leu Ala Ala Ala Gly Val Gln Pro His Ser Leu Asp Leu 245 250 255 Ser His Asn Ser Leu Arg Ala Thr Val Asn Pro Ser Ala Pro Arg Cys 260 265 270 Met Trp Ser Ser Ala Leu Asn Ser Leu Asn Leu Ser Phe Ala Gly Leu 275 280 285 Glu Gln Val Pro Lys Gly Leu Pro Ala Lys Leu Arg Val Leu Asp Leu 290 295 300 Ser Cys Asn Arg Leu Asn Arg Ala Pro Gln Pro Asp Glu Leu Pro Glu 305 310 315 320 Val Asp Asn Leu Thr Leu Asp Gly Asn Pro Phe Leu Val Pro Gly Thr 325 330 335 Ala Leu Pro His Glu Gly Ser Met Asn Ser Gly Val Val Pro Ala Cys 340 345 350 Ala Arg Ser Thr Leu Ser Val Gly Val Ser Gly Thr Leu Val Leu Leu 355 360 365Gln Gly Ala Arg Gly Phe Ala 370 375

Claims (20)

A composition for diagnosing brain cancer comprising an agent for measuring the expression level of CD169 (Cluster of Differentiation 169) protein or a gene encoding the same,
The expression level of the protein or a gene encoding it is measured for macrophages in brain tissue, the composition. The method of claim 1,
One or more proteins selected from the group consisting of CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1), and CD14 (Cluster of Differentiation 14) or genes encoding the same A composition comprising an agent for measuring expression levels. The method of claim 1,
The macrophages are derived from monocytes, the composition. The method of claim 1,
The agent for measuring the expression level of the protein comprises at least one selected from the group consisting of an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the protein, Composition . The method of claim 1,
The agent for measuring the expression level of the gene encoding the protein comprises at least one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the gene, composition. delete The method of claim 1,
The brain cancer is glioblastoma, the composition. A brain cancer diagnostic kit comprising the diagnostic composition of claim 1. CD169 (Cluster of Differentiation 169) CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1) and CD14 (Cluster of Differentiation 14 ) comprising the step of measuring the expression level of one or more proteins selected from the group consisting of or a gene encoding the same, as an information providing method for brain cancer diagnosis,
The biological sample is brain tissue,
The method, wherein the expression level of the protein or gene is measured for macrophages in the brain tissue. delete 10. The method of claim 9,
The method of claim 1, wherein the macrophages are CD169+CD14+F4/80low macrophages. delete 10. The method of claim 9,
The macrophages are derived from monocytes, the method. delete 10. The method of claim 9,
The method of claim 1, wherein the brain cancer is glioblastoma. A pharmaceutical composition for preventing or treating brain cancer, comprising as an active ingredient a macrophage comprising CD169 protein or an extracellular domain thereof. 17. The method of claim 16,
The macrophages are one or more proteins selected from the group consisting of CD45 (Cluster of Differentiation 45), F4/80 (EGF-like module-containing mucin-like hormone receptor-like 1), and CD14 (Cluster of Differentiation 14), or one or more proteins thereof. A pharmaceutical composition, further comprising an extracellular domain. 17. The method of claim 16,
The macrophages are derived from monocytes, pharmaceutical composition. delete 17. The method of claim 16,
The brain cancer is glioblastoma, a pharmaceutical composition.

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