KR20220015265A - Coposition for diagnosing infectious inflammatory disease comprising agents detecting the expression level of Wnt11 - Google Patents

Abstract

The present invention relates to: a composition for diagnosing the severity of infectious diseases, comprising an agent for measuring the expression level of Wnt family member 11 (Wnt11); and a pharmaceutical composition for preventing or treating infectious diseases, comprising a Wnt11 protein as an active ingredient. According to the present invention, it is possible to very accurately diagnose the severity of infectious diseases, which can be very effectively used in the establishment of treatment strategies for patients with infectious diseases.

Description

Composition for diagnosing infectious disease severity comprising agents detecting the expression level of Wnt11

The present invention relates to a composition for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11), and to a pharmaceutical composition for preventing or treating an infectious disease comprising the Wnt11 protein as an active ingredient.

Infectious diseases are diseases that occur when foreign substances such as bacteria, bacteria, and viruses appear and begin to live in blood, body fluids and tissues. . The prevalence of infectious diseases seems to decrease with the improvement of hygiene standards, but it is fatal due to the misuse of antibiotic administration, the increase in the use of immunosuppressants following transplantation, the decrease in immunity due to chemotherapy, and the increase in the number of people with underlying diseases such as diabetes and high blood pressure. The threat of infectious diseases with consequences is increasing.

In particular, most infectious diseases are accompanied by an inflammatory reaction at the site of infection, and some of them cause a systemic inflammatory reaction, which can lead to fatal results. In addition, since infectious patients due to infection can die, it is important to start appropriate antibiotic treatment as soon as possible, so accurate diagnosis and prediction of severity are essential for survival of infectious patients.

Meanwhile, a novel human coronavirus pandemic has recently spread rapidly around the world, posing a threat to global health. Surprisingly, the symptoms observed in patients with SARS-CoV-2 infection are similar to previously reported other viral infections such as severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). Many of these patients had pneumonia-related symptoms, such as acute respiratory distress syndrome (ARDS), cytokine secretion syndrome (CRS), multiorgan failure (MOF), and sepsis. Although many studies are being conducted to rapidly diagnose SARS-CoV-2 infection, no method for accurately diagnosing the severity of the disease has yet been reported.

Prediction of the severity of various infectious diseases, including SARS-CoV-2, is essential in establishing a patient's treatment strategy. can be important

Accordingly, the present inventors, while conducting a study on the relationship between an infectious disease and a Wnt family member, found that the expression level of Wnt11, or Wnt11 and Wnt5a in a biological sample from an infectious disease patient is very closely related to the severity of the disease, and The present invention has been completed.

Accordingly, an object of the present invention is to provide a composition for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

Another object of the present invention is to provide a kit for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

Another object of the present invention is to provide information necessary for diagnosing the severity of an infectious disease, (a) measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA in a biological sample provided from a patient suspected of an infectious disease ; (b) to provide a method for detecting Wnt11 protein or mRNA, comprising the step of comparing the expression level of the Wnt11 protein or mRNA with a normal person.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating an infectious disease comprising a Wnt11 (Wnt family member 11) protein, a vector comprising a polynucleotide encoding the Wnt11 protein, or a cell comprising the vector as an active ingredient will do

In order to achieve the object of the present invention, the present invention provides a composition for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

In order to achieve another object of the present invention, the present invention provides a kit for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

In order to achieve another object of the present invention, in order to provide information necessary for diagnosing the severity of an infectious disease, (a) Wnt11 (Wnt family member 11) protein or mRNA in a biological sample provided from a patient suspected of an infectious disease measuring the expression level; (b) provides a method for detecting Wnt11 protein or mRNA, comprising the step of comparing the expression level of the Wnt11 protein or mRNA with a normal person.

In order to achieve another object of the present invention, the present invention provides a Wnt11 (Wnt family member 11) protein, a vector containing a polynucleotide encoding the Wnt11 protein, or an infectious disease prevention or treatment comprising a cell containing the vector as an active ingredient A pharmaceutical composition is provided.

Hereinafter, the present invention will be described in detail.

According to an embodiment of the present invention, the expression levels of Wnt11 protein and mRNA in blood and peripheral blood mononuclear cells (PBMC) isolated from patients with infectious diseases significantly increase, but as the severity of the disease increases, the expression level of Wnt11 protein and mRNA was found to decrease. Therefore, it was confirmed that only a simple blood test can very accurately diagnose the severity of an infectious disease.

Accordingly, the present invention provides a composition for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

In the present invention, the Wnt11 is encoded by the WNT11 gene in humans. The amino acid sequence and mRNA base sequence of the Wnt11 protein are known as Genebank accession numbers NP_004617 (protein), NM_004626 (mRNA), and the like, and preferably SEQ ID NO: 1 (protein) or SEQ ID NO: 2 (mRNA). It may be represented.

In the present invention, 'expression' means that a protein or nucleic acid is produced in a cell. 'Protein' is used interchangeably with 'polypeptide' or 'peptide' and refers to, for example, a polymer of amino acid residues as commonly found in proteins in their natural state. 'Polynucleotide' or 'nucleic acid' refers to deoxyribonucleotides (DNA) or ribonucleotides (RNA) in single- or double-stranded form. Unless otherwise limited, known analogs of natural nucleotides that hybridize to nucleic acids in a manner analogous to naturally occurring nucleotides are also included. 'mRNA' is an RNA that delivers genetic information (gene-specific nucleotide sequence) to the ribosome, which specifies the amino acid sequence from a specific gene during protein synthesis.

By 'diagnosing' is meant ascertaining the presence or character of a pathological condition. Diagnosis in the present invention means a diagnosis of the severity of an infectious disease, and the 'severity' refers to the seriousness of a patient with an infectious disease, specifically, the presence of complications, the degree of risk of death, whether accompanied by severe acute respiratory syndrome It may mean etc.

In one embodiment of the present invention, the agent for measuring the expression level of the Wnt11 protein may be an antibody, antibody fragment, or aptamer that specifically binds to the Wnt11 protein.

The term 'antibody' refers to an immunoglobulin that specifically binds to an antigenic site. The antibody in the present invention does not react with other types of proteins other than Wnt11, and is an antibody that specifically binds to Wnt11 protein. The Wnt11 antibody can be prepared by cloning the Wnt11 gene into an expression vector to obtain a protein encoded by the gene, and from the obtained protein according to a conventional method in the art. A Wnt11 protein-specific antibody can also be prepared using a fragment of the Wnt11 protein containing the Wnt11 antigenic region.

The form of the antibody of the present invention is not particularly limited, and includes a polyclonal antibody or a monoclonal antibody. In addition, as long as it has antigen-antibody binding properties, a part of the whole antibody is also included in the antibody of the present invention, and all types of immunoglobulin antibodies that specifically bind to Wnt11 are included. For example, a functional fragment of an antibody molecule as well as a complete antibody having two full-length light chains and two full-length heavy chains, i.e. Fab, F(ab'), F(ab') with antigen-binding function. 2 and Fv, and the like. Furthermore, the antibody of the present invention includes special antibodies such as humanized antibodies and chimeric antibodies and recombinant antibodies as long as they can specifically bind to Wnt11 protein.

In the present invention, the Wnt11 protein preferably includes the human Wnt11 amino acid sequence shown in SEQ ID NO: 1, and the antibody specifically binding to the Wnt11 protein in the present invention preferably comprises the amino acid sequence shown in SEQ ID NO: 1 It may be an antibody that specifically binds to a protein having a The diagnostic composition of the present invention comprising the Wnt11-specific antibody as an agent for measuring the expression level of Wnt11 may additionally include an agent necessary for a method for detecting a known protein, and the known protein is detected using the composition Any method can be used without limitation to measure the level of Wnt11 protein.

In the present invention, the 'aptamer' refers to single-stranded DNA (ssDNA) or RNA having high specificity and affinity for a specific substance. Aptamers have very high affinity for specific substances and are stable, can be synthesized by a relatively simple method, can be modified in various ways to increase binding strength, and can be targeted to cells, proteins, and even small organic substances. Its specificity and stability are very high compared to antibodies that have already been developed. In the present invention, the type and form of the aptamer is not particularly limited as long as it can bind to Wnt11.

In another embodiment of the present invention, the agent for measuring the expression level of Wnt11 mRNA may be a primer pair, a probe, or a combination thereof that specifically binds to Wnt11 mRNA.

The Wnt11 mRNA may be derived from mammals including humans, and may preferably include the human Wnt11 mRNA base sequence represented by SEQ ID NO: 2. The diagnostic composition of the present invention comprising the Wnt11 mRNA-specific probe or primer set as an agent for measuring the expression level of Wnt11 may further include an agent required for a known RNA detection method. The level of Wnt11 mRNA in a subject can be measured using the present composition using any known method for detecting RNA without limitation.

The 'primer' is a short single-stranded oligonucleotide serving as a starting point of DNA synthesis. The primer specifically binds to a polynucleotide as a template under suitable buffer and temperature conditions, and DNA polymerase adds a nucleoside triphosphate having a base complementary to the template DNA to the primer to link it. is synthesized A primer generally consists of a sequence of 15 to 30 bases, and the melting temperature (Tm) at which it binds to the template strand varies depending on the base composition and length.

The sequence of the primer does not need to have a completely complementary sequence to a partial nucleotide sequence of the template, and it is sufficient if it hybridizes with the template and has sufficient complementarity within a range capable of performing an intrinsic function of the primer. Therefore, in the present invention, the primer for measuring the expression level of Wnt11 mRNA does not need to have a perfectly complementary sequence to the Wnt11 gene sequence, and the amount of Wnt11 mRNA is determined by amplifying a specific section of Wnt11 mRNA or Wnt11 cDNA through DNA synthesis. It is sufficient if it has a length and complementarity suitable for the purpose of measurement. The primer for the amplification reaction consists of a set (pair) that complementarily binds to the template (or sense) and the opposite side (antisense) of both ends of a specific section of Wnt11 mRNA to be amplified. Primers can be easily designed by those skilled in the art by referring to the Wnt11 mRNA or cDNA sequence.

In the present invention, the primers may preferably be a set or a pair that specifically binds to the Wnt11 mRNA base sequence represented by SEQ ID NO: 2.

The 'probe' refers to a fragment of a polynucleotide such as RNA or DNA having a length of several to several hundred base pairs, which can specifically bind to mRNA or cDNA (complementary DNA) of a specific gene. Meaning, it is labeled (labeling), so that the presence or absence of the target mRNA or cDNA to be bound, the amount of expression, etc. can be confirmed. For the purpose of the present invention, a probe complementary to Wnt11 mRNA may be used for diagnosing an infectious disease by performing a hybridization reaction with a sample of a subject and measuring the expression level of Wnt11 mRNA. Probe selection and hybridization conditions can be appropriately selected according to techniques known in the art.

In the present invention, the primer or probe may be chemically synthesized using a phosphoramidite solid support synthesis method or other well-known methods. In addition, the primer or probe can be variously modified according to methods known in the art within the range that does not interfere with hybridization with Wnt11 mRNA. Examples of such modifications include methylation, encapsulation, substitution of one or more homologues of natural nucleotides, and modifications between nucleotides, such as uncharged linkages such as methyl phosphonates, phosphotriesters, phosphoroamidates, carbamates, etc. ) or charged linkages (eg, phosphorothioate, phosphorodithioate, etc.), and fluorescence or enzymatic binding of a labeling material.

In one aspect of the present invention, the composition may further comprise an agent for measuring the expression level of Wnt5a protein or mRNA.

In the present invention, the Wnt5a is encoded by the WNT5A gene in humans. The amino acid sequence and mRNA base sequence of the Wnt5a protein are known as Genebank accession numbers NP_001243034 (protein), NM_001256105 (mRNA), and the like, and preferably SEQ ID NO: 3 (protein) or SEQ ID NO: 4 (mRNA). It may be represented.

According to an embodiment of the present invention, as a result of analyzing the expression levels of Wnt11 and Wnt5a proteins or mRNA in blood isolated from an infectious disease patient, the milder the disease, the higher the Wnt11/Wnt5a value, and the more severe the disease. It was confirmed that the lower the Wnt11/Wnt5a value.

Therefore, if the ratio is calculated by measuring the expression levels of Wnt11 and Wnt5a proteins or mRNA, the severity of an infectious disease can be predicted more accurately.

In the present invention, the term 'infection' means that one or more types of exogenous bacteria (including bacteria, gram-negative or gram-positive bacteria), viruses, and fungi (bacteria) invade the body and become settled, proliferated, and parasitic. The infectious disease may be any disease that occurs by causing a reaction in the living body as a result of infection by a pathogen. Reactions resulting from infectious diseases may include inflammation, pain, fever, fatigue, edema, and lowering of blood pressure.

In the present invention, the infectious disease may be preferably an infectious inflammatory disease, more preferably sepsis, septic shock, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, Severe acute respiratory syndrome coronavirus (SARS-CoV) infection, Middle East Respiratory Syndrome (MERS), salmonellosis, food poisoning, typhoid, paratyphoid, systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome ( multiple organ dysfunction syndrome, MODS), pneumonia, pulmonary tuberculosis, tuberculosis, cold, influenza, respiratory tract infection, rhinitis, nasopharyngitis, otitis media, bronchitis, lymphadenitis, parotitis, lymphadenitis, stomatitis, stomatitis, arthritis, myositis, dermatitis, vasculitis, gingivitis , periodontitis, keratitis, conjunctivitis, wound infection, peritonitis, hepatitis, osteomyelitis, cellulitis, meningitis, encephalitis, brain abscess, encephalomyelitis, meningitis, osteomyelitis, nephritis, carditis, endocarditis, enteritis, gastritis, esophagitis, duodenitis, colitis, urinary tract Inflammation, cystitis, vaginitis, cervicitis, salpingitis, erythematosus, bacterial dysentery, abscesses and ulcers, bacteremia, diarrhea, dysentery, gastroenteritis, gastroenteritis, genitourinary abscesses, open wounds or infections of wounds, purulent inflammation, abscesses, boils, Pyoderma, impetigo, folliculitis, cellulitis, post-operative wound infection, skin laceration syndrome, skin burn syndrome, thrombotic thrombocytopenia, hemolytic uremic syndrome, renal failure, pyelonephritis, glomerulonephritis, nervous system abscess, otitis media, sinusitis, pharyngitis, tonsillitis, mastitis prostatitis, cellulitis, gingivitis, dacryocystitis, pleurisy, abdominal abscess, liver abscess, cholecystitis, splenic abscess, pericarditis, myocarditis, placenta, amnioticitis, mastitis, mastitis, puerperal fever, toxic shock syndrome, Lyme disease, gas necrosis, atherosclerosis, mycobacterium avium syndrome (MAC), enterohaemorrhagic E. coli (EHEC) infection, enteropathogenic E. coli (E) PEC) infection, intestinal invasive E. coli infection (EIEC), methicillin-resistant Staphylococcus aureus (MRSA) infection, vancomycin-resistant Staphylococcus aureus (VRSA) infection, and may be at least one selected from the group consisting of listerosis, Most preferably, it may be sepsis, septic shock, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, but is not limited thereto.

In the present invention, the 'septicemia' is a systemic inflammatory reaction syndrome that appears as a complication of an infectious disease. It progresses to multiple organ dysfunction syndrome (MODS), disseminated intravascular coagulation syndrome (DIC), acute respiratory urgency syndrome (ARDS), or acute renal failure (AKI) resulting in liver and circulatory dysfunction It is a fatal disease that can cause death.

In the present invention, the sepsis is sepsis associated with the final stage of sepsis, severe sepsis, septic shock and multiple organ dysfunction syndrome accompanying sepsis (MODS), disseminated intravascular coagulation syndrome (DIC), All stages of sepsis, including but not limited to, the development of acute respiratory distress syndrome (ARDS) or acute renal failure (AKI).

In the present invention, the severe acute respiratory syndrome virus 2 is an RNA virus belonging to Corona, SARS-CoV-2, COVID-19, Covid-19, Corona-19, 2019-nCoV, 2019 novel coronavirus, coronavirus disease 2019, etc. is called In the present invention, the SARS-CoV-2 may be mutated by causing various mutations during infection, and these variants may also be included in the SARS-CoV-2 of the present invention.

The present invention also provides a kit for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.

The diagnostic kit of the present invention includes an antibody, an antibody fragment, an aptamer, or a primer that recognizes Wnt11 mRNA as a marker, which selectively recognizes Wnt11 protein as a marker in order to measure the expression level of Wnt11 as well as primers and probes suitable for analysis methods. or more other component compositions, solutions or devices may be included.

In a specific embodiment, the diagnostic kit may be a diagnostic kit comprising essential elements necessary for performing a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes each primer pair specific for a marker gene. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of each marker gene, and has a length of about 7 bp to 50 bp, more preferably 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 another aspect, it may be a diagnostic kit comprising 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.

Most preferably, it may be a diagnostic kit comprising essential elements necessary for performing ELISA. The ELISA kit contains an antibody specific for a marker protein. Antibodies are antibodies with high specificity and affinity for each 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 (in the form of conjugated with an antibody) and substrates thereof or capable of binding the antibody. other materials and the like. In addition, the kit of the present invention may include a washing solution or eluent capable of retaining only the bound protein marker while removing the substrate to be subjected to a color reaction with the enzyme, unbound protein, and the like.

The present invention also provides information necessary for diagnosing the severity of an infectious disease, comprising the steps of: (a) measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA in a biological sample provided from a patient suspected of an infectious disease; (b) provides a method for detecting Wnt11 protein or mRNA, comprising the step of comparing the expression level of the Wnt11 protein or mRNA with a normal person.

The present inventors first discovered that Wnt11 can function as a marker for diagnosing the severity of an infectious disease, thereby providing a method for providing information necessary for diagnosing the severity of an infectious disease by measuring the expression level of Wnt11. Hereinafter, the method of the present invention will be described step by step.

(a) measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA in a biological sample provided from a patient suspected of an infectious disease;

The biological sample can be used without limitation as long as it is collected from a subject to diagnose the severity of an infectious disease, for example, cells or tissues, blood, plasma, serum, saliva, nasal fluid, sputum, joint capsule fluid obtained by biopsy, amniotic fluid, ascites, cervical or vaginal secretions, urine and cerebrospinal fluid, and the like. Preferably, it may be blood, plasma, or serum.

Alternatively, the biological sample in step (a) may be collected from a subject that has already been determined to have an infectious disease by other means.

The level of the Wnt11 protein can be detected or measured using an antibody that specifically binds to the Wnt11 protein. The Wnt11 protein-specific antibody is as described in the diagnostic composition of the present invention. Methods for measuring the expression level of Wnt11 protein can be used without limitation, methods known in the art, for example, western blotting (western blotting), dot blotting (dot blotting), enzyme-linked immunosorbent assay (enzyme-linked immunosorbent) assay, ELISA), radioimmunoassay (RIA), radioimmunodiffusion, octeroni immunodiffusion, rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation, complement fixation assay, flow cytometry (FACS) or There is a protein chip method and the like, but is not limited thereto. Preferably, an ELISA method can be used.

The level of Wnt11 mRNA is determined by amplifying Wnt11 mRNA or cDNA from a sample of a subject using a primer set or a probe that specifically binds to Wnt11 mRNA, or by using a probe and hybridization to determine the level of Wnt11 mRNA in the sample. The presence and level of expression can be measured. Primers and probes of Wnt11 are the same as those described in the diagnostic composition of the present invention. Measurement of the Wnt11 mRNA expression level can be used without limitation by a method for confirming the expression level conventional in the art, and examples of the analysis method include reverse transcription polymerase chain reaction (RT-PCR), competitive RT-PCR (competitive RT). -PCR), real-time RT-PCR, RNase protection assay (RPA), northern blotting, DNA microarray chip, RNA sequencing ( RNA sequencing), a hybridization method using a nanostring, an in situ hybridization method of a tissue section, and the like, but are not limited thereto.

(b) comparing the expression level of the Wnt11 protein or mRNA with a normal person;

The Wnt11 expression level of the subject measured by the method of step (a) described above is compared with the Wnt11 level of a normal person measured by the same method. As the expression level of Wnt11 is higher than that of a healthy normal person, it can be determined that the severity of the infectious disease of the subject is lower.

In a preferred embodiment of the present invention, the expression level of Wnt5a protein or mRNA is further measured in step (a), and the lower the Wnt11/Wnt5a value compared to the normal in step (b), the higher the severity of the disease, and Wnt11/ It may be characterized in that the higher the Wnt5a value, the lower the severity of the disease.

After accurately determining the severity of an infectious disease according to the method of the present invention, it may be possible to establish a treatment strategy for the patient according to the severity of the disease.

The present invention also provides a Wnt11 (Wnt family member 11) protein, a vector comprising a polynucleotide encoding the Wnt11 protein, or a pharmaceutical composition for preventing or treating an infectious disease comprising a cell comprising the vector as an active ingredient.

In the present invention, the Wnt11 protein should be interpreted as including a variant or functional equivalent thereof. The protein variant or functional equivalent refers to a Wnt11 protein of the present invention having 60%, preferably 70%, more preferably 80% or more sequence homology to the amino acid sequence of the Wnt11 protein as a result of addition, substitution or deletion of amino acids. Refers to a polypeptide that exhibits substantially the same activity as the protein. The functional equivalent may include, for example, an amino acid sequence variant in which some of the amino acids of the amino acid sequence of the Wnt11 protein are substituted, deleted, or added. The substitution of amino acids may preferably be conservative substitutions, and examples of conservative substitutions of naturally occurring amino acids are as follows; Aliphatic amino acids (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn) ) and sulfur-containing amino acids (Cys, Met). Deletion of amino acids may be preferably located at a portion not directly involved in the activity of the Wnt11 protein of the present invention. In addition, the scope of the functional equivalent may include polypeptide derivatives in which some chemical structures of the polypeptide are modified while maintaining the basic backbone of the Wnt11 protein and its physiological activity. For example, a fusion protein made by fusion with another protein while maintaining a structural change and physiological activity to change the stability, storage, volatility or solubility of the Wnt11 protein of the present invention may be included therein.

In addition, the Wnt11 protein according to the present invention may be used by itself or in the form of a pharmaceutically acceptable salt. "Pharmaceutically acceptable" in the present invention means that it is physiologically acceptable, does not inhibit the action of the active ingredient when administered to humans, and does not usually cause allergic reactions such as gastrointestinal disorders, dizziness, or similar reactions. . The salt is preferably an acid addition salt formed with a pharmaceutically acceptable free acid, and an organic acid and an inorganic acid can be used as the free acid. The organic acid is not limited thereto, but citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid and aspartic acid. In addition, the inorganic acid includes, but is not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid.

In the present invention, the Wnt11 protein or fragment thereof can be extracted from nature or synthesized (Merrifeld, J. Amer. chem. Soc. 85:2149-2156, 1963), or prepared by a genetic recombination method based on a DNA sequence. (Sambrook et al, Molecular Cloning, Cold Spring Harbor Laboratory Press, New York, USA, 2nd ed., 1989).

In the present invention, the vector may be selected from the group consisting of linear DNA, plasmid DNA, and recombinant viral vectors, and the virus is selected from the group consisting of retroviruses, adenoviruses, adeno-associated viruses, herpes simplex viruses and lentiviruses. can be

Cells containing the vector are preferably selected from the group consisting of hematopoietic stem cells (hematopoietic stem cells), dendritic cells (dendritic cells), autologous tumor cells (autologous tumor cells) and established tumor cells (established tumor cells) , but not limited thereto.

The composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. can

For suitable formulations known in the art, reference may be made to those disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA.

Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc. mixed with the extract. is prepared In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

As used herein, the term “administration” means providing a given composition of the present invention to a subject by any suitable method.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the individual, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. For a desirable effect, the composition of the present invention may be administered at 0.001 to 1000 mg/kg per day. 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.

When a vector comprising a polynucleotide encoding a Wnt11 protein or a fragment thereof is included in the pharmaceutical composition of the present invention, it is preferably contained in an amount of 0.05 to 500 mg, more preferably 0.1 to 300 mg, and In the case of a recombinant virus comprising a polynucleotide encoding a protein, it may be preferable to contain 10 ³ to 10 ¹² IU, but is not limited thereto.

In addition, when the cell containing the polynucleotide encoding the protein is included in the pharmaceutical composition of the present invention, it is preferable to contain 10 ³ to 10 ⁸ such cells, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered to an individual by various routes. Any mode of administration can be envisaged, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebrovascular injection.

According to the present invention, it is possible to very accurately diagnose the severity of an infectious disease, which can be very usefully utilized in establishing a treatment strategy for an infectious disease patient.

1 shows 20 normal controls, 80 SARS-CoV-2 infection patients, 25 SARS-CoV-2 patients with severe respiratory syndrome (SARS-CoV-2 ARDS), and 25 discharged SARS-CoV-2 patients. It is a diagram showing clinical information of an infectious disease patient.
2 shows 20 normal controls, 80 SARS-CoV-2 infected patients, 25 SARS-CoV-2 patients with severe respiratory syndrome (SARS-CoV-2 ARDS), and 25 discharged SARS-CoV-2 patients. It is a result of measuring the protein or mRNA expression levels of Wnt5a and Wnt11 in the plasma of an infected patient (discharged).
3 shows Wnt5a or Wnt11 proteins in PBMCs isolated from patients with SARS-CoV-2 infection, SARS-CoV-2 patients with severe respiratory syndrome (SARS-CoV-2 ARDS), and discharged patients with SARS-CoV-2 infection. is a result of measuring the expression level of
Figure 4 shows the indicated cytokines after treatment with anti-Wnt5a antibody (Wnt5a ab) or recombinant human Wnt11 protein (Wnt11) in PBMCs of SARS-CoV-2 patients with severe respiratory syndrome (SARS-CoV-2 ARDS). is the result of evaluating the expression level of

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Experimental method

Whole blood was collected from a patient with SARS-CoV-2 infection admitted to Yeungnam University Hospital. The patient admitted to the hospital did not take any medications. The study protocol was approved by the Daegu Yeungnam University Hospital Clinical Trial Review Committee.

The concentrations of Wnt5a, Wnt11 or cytokines in the plasma of patients with SARS-CoV-2 infection were quantified using a commercially available ELISA kit according to the manufacturer's instructions. Human recombinant WNT11 protein (H00007481-P01, Abnova), anti-Wnt5a antibody (MAB645, R&D Systems), anti-Wnt11 antibody (ab31962, Abcam), human protein Wnt-5a ELISA kit (MBS2886311, MyBioSource) and human protein Wnt- 11 ELISA kit (MBS281148, MyBioSource) was used.

Heparinized blood samples were used fresh within 4 hours, and peripheral blood mononuclear cells (PBMCs) were isolated from blood using Ficoll-Hypaque or NycoPrep. Then, purified PBMCs were obtained using the MACSprep™ PBMC isolation kit. To verify the effect of Wnt5a neutralizing antibody or recombinant human Wnt11 on the inhibition of cytokine secretion and NF-βB activation, PBMCs isolated from SARS-CoV-2 infected patients were treated with Wnt5a antibody (20 μg/ml) or recombinant human Wnt11 (10 ng/ml) and incubated for 6 hours. Supernatants were used for analysis of cytokine ELISA, and lysates were used for NF-κB activity assay by ELISA-based NF-κB family transcription factor assay kit (43296; Active Motif). All experiments were performed independently at least three times.

Experiment result

In order to establish a reliable diagnostic biomarker, the present inventors conducted a preliminary study by exploring clinical symptoms and various risk factors for patients with severe SARS-CoV-2 infection admitted to Yeungnam University College of Medicine. We conducted a study to discover novel biomarkers in blood based on patient information such as age, BMI, and comorbidities (Fig. 1A).

Plasma samples from patients with SARS-CoV-2 infection were divided according to disease severity; Normal subjects (control group), SARS-CoV-2 patients, SARS-CoV-2 patients with acute respiratory distress syndrome (SARS-CoV-2 ARDS), and SARS-CoV-2 infectious patients discharged from hospital (discharged).

As a result of the experiment, as shown in FIG. 2 , there was a slight difference in the level of Wnt5a secretion between mild SARS-CoV2 infection and the control group. However, the Wnt5a protein level was significantly increased in the blood of SARS-CoV-2 ARDS. Interestingly, Wnt5a protein levels decreased again in discharged patients (Fig. 2a).

In addition, although Wnt5a levels were kept low in the plasma of surviving patients, high levels of Wnt5a were still observed in the dead patients, indicating a correlation between Wnt5 levels and disease severity ( FIG. 2b ).

Conversely, Wnt11 protein levels were strongly induced in the plasma of mild SARS-CoV2 patients and discharged individuals, but remained at normal levels in SARS-CoV-2 ARDS patients (Fig. 2c).

Changes in the expression levels of Wnt5a and Wnt11 in patients with SARS-CoV-2 infection were confirmed to show a similar trend in PBMCs ( FIGS. 3A and 3B ).

The effects of Wnt5a and Wnt11 on the inflammatory response in patients with SARS-CoV-2 infection were further confirmed. PBMCs from patients with SARS-CoV-2 infection were treated with anti-Wnt5a neutralizing antibody or recombinant Wnt11 (rWnt11) protein. NF-κB activation assay showed that treatment with anti-Wnt5a antibody did not significantly reduce the inflammatory response in PBMCs of SARS-CoV-2 ARDS (Fig. 4a). Moreover, secretion of various cytokines including IL-6, IL-1β, IL-4, IFN-β and TNF-β was not significantly changed upon treatment with anti-Wnt5a antibody ( FIGS. 4B-4G ). However, treatment of rWnt11 on PBMCs of SARS-CoV-2 ARDS showed a dramatic inhibitory effect on NF-κB activation and cytokine production ( FIGS. 4B-4G ). These results suggest that, while Wnt11 protein has great efficacy in reducing the inflammatory response induced by SARS-CoV-2 infection, the possibility of Wnt5a as a potential therapeutic target is low.

According to the present invention, it is possible to diagnose the severity of an infectious disease very accurately, and it can be very usefully utilized in establishing a treatment strategy for patients with an infectious disease, and thus industrial applicability is very high.

<110> Kyungpook National University Industry-Academic Cooperation Foundation KYUNGPOOK NATIONAL UNIVERSITY HOSPITAL <120> Composition for diagnosing infectious inflammatory disease comprising agents detecting the expression level of Wnt11 <130> NP20-0058 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 354 <212> PRT <213> Artificial Sequence <220> <223> Human Wnt11 protein amino acid sequence <400> 1 Met Arg Ala Arg Pro Gln Val Cys Glu Ala Leu Leu Phe Ala Leu Ala 1 5 10 15 Leu Gln Thr Gly Val Cys Tyr Gly Ile Lys Trp Leu Ala Leu Ser Lys 20 25 30 Thr Pro Ser Ala Leu Ala Leu Asn Gln Thr Gln His Cys Lys Gln Leu 35 40 45 Glu Gly Leu Val Ser Ala Gln Val Gln Leu Cys Arg Ser Asn Leu Glu 50 55 60 Leu Met His Thr Val Val His Ala Ala Arg Glu Val Met Lys Ala Cys 65 70 75 80 Arg Arg Ala Phe Ala Asp Met Arg Trp Asn Cys Ser Ser Ile Glu Leu 85 90 95 Ala Pro Asn Tyr Leu Leu Asp Leu Glu Arg Gly Thr Arg Glu Ser Ala 100 105 110 Phe Val Tyr Ala Leu Ser Ala Ala Ala Ile Ser His Ala Ile Ala Arg 115 120 125 Ala Cys Thr Ser Gly Asp Leu Pro Gly Cys Ser Cys Gly Pro Val Pro 130 135 140 Gly Glu Pro Pro Gly Pro Gly Asn Arg Trp Gly Gly Cys Ala Asp Asn 145 150 155 160 Leu Ser Tyr Gly Leu Leu Met Gly Ala Lys Phe Ser Asp Ala Pro Met 165 170 175 Lys Val Lys Lys Thr Gly Ser Gln Ala Asn Lys Leu Met Arg Leu His 180 185 190 Asn Ser Glu Val Gly Arg Gln Ala Leu Arg Ala Ser Leu Glu Met Lys 195 200 205 Cys Lys Cys His Gly Val Ser Gly Ser Cys Ser Ile Arg Thr Cys Trp 210 215 220 Lys Gly Leu Gln Glu Leu Gln Asp Val Ala Ala Asp Leu Lys Thr Arg 225 230 235 240 Tyr Leu Ser Ala Thr Lys Val Val His Arg Pro Met Gly Thr Arg Lys 245 250 255 His Leu Val Pro Lys Asp Leu Asp Ile Arg Pro Val Lys Asp Ser Glu 260 265 270 Leu Val Tyr Leu Gln Ser Ser Pro Asp Phe Cys Met Lys Asn Glu Lys 275 280 285 Val Gly Ser His Gly Thr Gln Asp Arg Gln Cys Asn Lys Thr Ser Asn 290 295 300 Gly Ser Asp Ser Cys Asp Leu Met Cys Cys Gly Arg Gly Tyr Asn Pro 305 310 315 320 Tyr Thr Asp Arg Val Val Glu Arg Cys His Cys Lys Tyr His Trp Cys 325 330 335 Cys Tyr Val Thr Cys Arg Arg Cys Glu Arg Thr Val Glu Arg Tyr Val 340 345 350 Cys Lys <210> 2 <211> 1743 <212> DNA <213> Artificial Sequence <220> <223> Human WNT11 gene sequence <400> 2 gaggcctggg tccccgtccg ctgtgcgacc tagggggctc ctcctctccc tgggagacca 60 ggctggacac agatcccccg ctgcggcgtg caggaccagc ggcggccgtg caggcggagg 120 acttcggcgc ggctcctcct gggtgtgacc ccgggcgcgc ccgccgcgcg acgatgaggg 180 cgcggccgca ggtctgcgag gcgctgctct tcgccctggc gctccagacc ggcgtgtgct 240 atggcatcaa gtggctggcg ctgtccaaga caccatcggc cctggcactg aaccagacgc 300 aacactgcaa gcagctggag ggtctggtgt ctgcacaggt gcagctgtgc cgcagcaacc 360 tggagctcat gcacacggtg gtgcacgccg cccgcgaggt catgaaggcc tgtcgccggg 420 cctttgccga catgcgctgg aactgctcct ccattgagct cgcccccaac tatttgcttg 480 acctggagag agggacccgg gagtcggcct tcgtgtatgc gctgtcggcc gccgccatca 540 gccacgccat cgcccgggcc tgcacctccg gcgacctgcc cggctgctcc tgcggccccg 600 tcccaggctc tgcgcgcctc tctggaaatg aagtgtaagt gccatggggt gtctggctcc 660 tgctccatcc gcacctgctg gaaggggctg caggagctgc aggatgtggc tgctgacctc 720 aagacccgat acctgtcggc caccaaggta gtgcaccgac ccatgggcac ccgcaagcac 780 ctggtgccca aggacctgga tatccggcct gtgaaggact cggaactcgt ctatctgcag 840 agctcacctg acttctgcat gaagaatgag aaggtgggct cccacgggac acaagacagg 900 cagtgcaaca agacatccaa cggaagcgac agctgcgacc ttatgtgctg cgggcgtggc 960 tacaacccct acacagaccg cgtggtcgag cggtgccact gtaagtacca ctggtgctgc 1020 tacgtcacct gccgcaggtg tgagcgtacc gtggagcgct atgtctgcaa gtgaggccct 1080 gccctccgcc ccacgcagga gcgaggactc tgctcaagga ccctcagcaa ctggggccag 1140 gggcctggag acactccatg gagctctgct tgtgaattcc agatgccagg catgggaggc 1200 ggcttgtgct ttgccttcac ttggaagcca ccaggaacag aaggtctggc caccctggaa 1260 ggagggcagg acatcaaagg aaaccgacaa gattaaaaat aacttggcag cctgaggctc 1320 tggagtgccc acaggctggt gtaaggagcg gggcttggga tcggtgagac tgatacagac 1380 ttgacctttc agggccacag agaccagcct ccgggaaggg gtctgcccgc cttcttcaga 1440 atgttctgcg ggaccccctg gcccaccctg gggtctgagc ctgctgggcc caccacatgg 1500 aatcactagc ttgggttgta aatgttttct tttgtttttt gctttttctt cctttgggat 1560 gtggaagcta cagaaatatt tataaaacat agctttttct ttggggtggc acttctcaat 1620 tcctctttat atattttata tatataaata tatatgtata tatataatga tctctatttt 1680 aaaactagct ttttaagcag ctgtatgaaa taaatgctga gtgagcccca gcccgcccct 1740 gca 1743 <210> 3 <211> 365 <212> PRT <213> Artificial Sequence <220> <223> Human Wnt5a protein amino acid sequence <400> 3 Met Ala Gly Ser Ala Met Ser Ser Lys Phe Phe Leu Val Ala Leu Ala 1 5 10 15 Ile Phe Phe Ser Phe Ala Gln Val Val Ile Glu Ala Asn Ser Trp Trp 20 25 30 Ser Leu Gly Met Asn Asn Pro Val Gln Met Ser Glu Val Tyr Ile Ile 35 40 45 Gly Ala Gln Pro Leu Cys Ser Gln Leu Ala Gly Leu Ser Gln Gly Gln 50 55 60 Lys Lys Leu Cys His Leu Tyr Gln Asp His Met Gln Tyr Ile Gly Glu 65 70 75 80 Gly Ala Lys Thr Gly Ile Lys Glu Cys Gln Tyr Gln Phe Arg His Arg 85 90 95 Arg Trp Asn Cys Ser Thr Val Asp Asn Thr Ser Val Phe Gly Arg Val 100 105 110 Met Gln Ile Gly Ser Arg Glu Thr Ala Phe Thr Tyr Ala Val Ser Ala 115 120 125 Ala Gly Val Val Asn Ala Met Ser Arg Ala Cys Arg Glu Gly Glu Leu 130 135 140 Ser Thr Cys Gly Cys Ser Arg Ala Ala Arg Pro Lys Asp Leu Pro Arg 145 150 155 160 Asp Trp Leu Trp Gly Gly Cys Gly Asp Asn Ile Asp Tyr Gly Tyr Arg 165 170 175 Phe Ala Lys Glu Phe Val Asp Ala Arg Glu Arg Glu Arg Ile His Ala 180 185 190 Lys Gly Ser Tyr Glu Ser Ala Arg Ile Leu Met Asn Leu His Asn Asn 195 200 205 Glu Ala Gly Arg Arg Thr Val Tyr Asn Leu Ala Asp Val Ala Cys Lys 210 215 220 Cys His Gly Val Ser Gly Ser Cys Ser Leu Lys Thr Cys Trp Leu Gln 225 230 235 240 Leu Ala Asp Phe Arg Lys Val Gly Asp Ala Leu Lys Glu Lys Tyr Asp 245 250 255 Ser Ala Ala Ala Met Arg Leu Asn Ser Arg Gly Lys Leu Val Gln Val 260 265 270 Asn Ser Arg Phe Asn Ser Pro Thr Thr Gln Asp Leu Val Tyr Ile Asp 275 280 285 Pro Ser Pro Asp Tyr Cys Val Arg Asn Glu Ser Thr Gly Ser Leu Gly 290 295 300 Thr Gln Gly Arg Leu Cys Asn Lys Thr Ser Glu Gly Met Asp Gly Cys 305 310 315 320 Glu Leu Met Cys Cys Gly Arg Gly Tyr Asp Gln Phe Lys Thr Val Gln 325 330 335 Thr Glu Arg Cys His Cys Lys Phe His Trp Cys Cys Tyr Val Lys Cys 340 345 350 Lys Lys Cys Thr Glu Ile Val Asp Gln Phe Val Cys Lys 355 360 365 <210> 4 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> Human Wnt5a gene sequence <400> 4 gctccactcg cctccgtgct cctctcgccc atggaattaa ttctggctcc acttgttgct 60 cggcccagaa gtccattgga atattaagcc caggagttgc tttggggatg gctggaagtg 120 caatgtcttc caagttcttc ctagtggctt tggccatatt tttctccttc gcccaggttg 180 taattgaagc caattcttgg tggtcgctag gtatgaataa ccctgttcag atgtcagaag 240 tatattattat aggagcacag cctctctgca gccaactggc aggactttct caaggacaga 300 agaaactgtg ccacttgtat caggaccaca tgcagtacat cggagaaggc gcgaagacag 360 gcatcaaaga atgccagtat caattccgac atcgaaggtg gaactgcagc actgtggata 420 acacctctgt ttttggcagg gtgatgcaga taggcagccg cgagacggcc ttcacatacg 480 cggtgagcgc agcaggggtg gtgaacgcca tgagccgggc gtgccgcgag ggcgagctgt 540 ccacctgcgg ctgcagccgc gccgcgcgcc ccaaggacct gccgcgggac tggctctggg 600 gcggctgcgg cgacaacatc gactatggct accgctttgc caaggagttc gtggacgccc 660 gcgagcggga gcgcatccac gccaagggct cctacgagag tgctcgcatc ctcatgaacc 720 tgcacaacaa cgaggccggc cgcaggacgg tgtacaacct ggctgatgtg gcctgcaagt 780 gccatggggt gtccggctca tgtagcctga agacatgctg gctgcagctg gcagacttcc 840 gcaaggtggg tgatgccctg aaggagaagt acgacagcgc ggcggccatg cggctcaaca 900 gccggggcaa gttggtacag gtcaacagcc gcttcaactc gcccaccaca caagacctgg 960 tctacatcga ccccagccct gactactgcg tgcgcaatga gagcaccggc tcgctgggca 1020 cgcagggccg cctgtgcaac aagacgtcgg agggcatgga tggctgcgag ctcatgtgct 1080 gcggccgtgg ctacgaccag ttcaagaccg tgcagacgga gcgctgccac tgcaagttcc 1140 actggtgctg ctacgtcaag tgcaagaagt gcacggagat cgtggaccag tttgtgtgca 1200 agtagtgggt gccacccagc actcagcccc gctcccagga cccgcttatt tatagaaagt 1260 acagtgattc tggtttttgg tttttagaaa tattttttat ttttccccaa gaattgcaac 1320 cggaaccatt ttttttcctg ttaccatcta agaactctgt ggtttattat taatattata 1380 attattattt ggcaataatg ggggtgggaa ccaagaaaaa tatttatttt gtggatcttt 1440 gaaaaggtaa tacaagactt cttttgatag tatagaatga aggggaaata acacataccc 1500 taacttagct gtgtggacat ggtacacatc cagaaggtaa agaaatacat tttctttttc 1560 tcaaatatgc catcatatgg gatgggtagg ttccagttga aagagggtgg tagaaatcta 1620 ttcacaattc agcttctatg accaaaatga gttgtaaatt ctctggtgca agataaaagg 1680 tcttgggaaa acaaaacaaa acaaaacaaa cctcccttcc ccagcagggc tgctagcttg 1740 ctttctgcat tttcaaaatg ataatttaca atggaaggac aagaatgtca tattctcaag 1800 gaaaaaaggt atatcacatg tctcattctc ctcaaatatt ccatttgcag acagaccgtc 1860 atattctaat agctcatgaa atttgggcag cagggaggaa agtccccaga aattaaaaaa 1920 tttaaaactc ttatgtcaag atgttgattt gaagctgtta taagaattag gattccagat 1980 tgtaaaaaga tccccaaatg attctggaca ctagattttt ttgtttgggg aggttggctt 2040 gaacataaat gaaaatatcc tgttattttc ttagggatac ttggttagta aattataata 2100 gtaaaaataa tacatgaatc ccattcacag gttctcagcc caagcaacaa ggtaattgcg 2160 tgccattcag cactgcacca gagcagacaa cctatttgag gaaaaacagt gaaatccacc 2220 ttcctcttca cactgagccc tctctgattc ctccgtgttg tgatgtgatg ctggccacgt 2280 ttccaaacgg cagctccact gggtcccctt tggttgtagg acaggaaatg aaacattagg 2340 agctctgctt ggaaaacagt tcactactta gggatttttg tttcctaaaa cttttatttt 2400 gaggagcagt agttttctat gttttaatga cagaacttgg ctaatggaat tcacagaggt 2460 gttgcagcgt atcactgtta tgatcctgtg tttagattat ccactcatgc ttctcctatt 2520 gtactgcagg tgtaccttaa aactgttccc agtgtacttg aacagttgca tttataaggg 2580 gggaaatgtg gtttaatggt gcctgatatc tcaaagtctt ttgtacataa catatatata 2640 tatatacata tatataaata taaatataaa tatatctcat tgcagccagt gatttagatt 2700 tacagtttac tctggggtta tttctctgtc tagagcattg ttgtccttca ctgcagtcca 2760 gttgggatta ttccaaaagt tttttgagtc ttgagcttgg gctgtggccc tgctgtgatc 2820 ataccttgag cacgacgaag caaccttgtt tctgaggaag cttgagttct gactcactga 2880 aatgcgtgtt gggttgaaga tatctttttt cttttctgcc tcaccccttt gtctccaacc 2940 tccatttctg ttcactttgt ggagagggca ttacttgttc gttatagaca tggacgttaa 3000 gagatattca aaactcagaa gcatcagcaa tgtttctctt ttcttagttc attctgcaga 3060 atggaaaccc atgcctatta gaaatgacag tacttattaa ttgagtccct aaggaatatt 3120 cagcccacta catagatagc tttttttttt ttttttttaa taaggacacc tctttccaaa 3180 cagtgccatc aaatatgttc ttatctcaga cttacgttgt tttaaaagtt tggaaagata 3240 cacatctttc atacccccct taggcaggtt ggctttcata tcacctcagc caactgtggc 3300 tcttaattta ttgcataatg atattcacat cccctcagtt gcagtgaatt gtgagcaaaa 3360 gatcttgaaa gcaaaaagca ctaattagtt taaaatgtca cttttttggt ttttattata 3420 caaaaaccat gaagtacttt ttttatttgc taaatcagat tgttcctttt tagtgactca 3480 tgtttatgaa gagagttgag tttaacaatc ctagctttta aaagaaacta tttaatgtaa 3540 aatattctac atgtcattca gatattatgt atatcttcta gcctttattc tgtactttta 3600 atgtacatat ttctgtcttg cgtgatttgt atatttcact ggtttaaaaa acaaacatcg 3660 aaaggcttat gccaaatgga agatagaata taaaataaaa cgttacttgt atattggtaa 3720 gtggtttcaa ttgtccttca gataattcat gtggagattt ttggagaaac catgacggat 3780 agtttaggat gactacatgt caaagtaata aaagagtggt gaattttacc aaaaccaagc 3840 tatttggaag cttcaaaagg tttctatatg taatggaaca aaaggggaat tctcttttcc 3900 tatatatgtt ccttacaaaa aaaaaaaaaa aagaaatcaa gcagatggct taaagctggt 3960 tataggattg ctcacattct tttagcatta tgcatgtaac ttaattgttt tagagcgtgt 4020 tgctgttgta acatcccaga gaagaatgaa aaggcacatg cttttatccg tgaccagatt 4080 tttagtccaa aaaaatgtat ttttttgtgt gtttaccact gcaactattg cacctctcta 4140 tttgaattta ctgtggacca tgtgtggtgt ctctatgccc tttgaaagca gtttttataa 4200 aaagaaagcc cgggtctgca gagaatgaaa actggttgga aactaaaggt tcattgtgtt 4260 aagtgcaatt aatacaagtt attgtgcttt tcaaaaatgt acacggaaat ctggacagtg 4320 ctccacagat tgatacatta gcctttgctt tttctctttc cggataacct tgtaacatat 4380 tgaaaccttt taaggatgcc aagaatgcat tattccacaa aaaaacagca gaccaacata 4440 tagagtgttt aaaatagcat ttctgggcaa attcaaactc ttgtggttct aggactcaca 4500 tctgtttcag tttttcctca gttgtatatt gaccagtgtt ctttattgca aaaacatata 4560 cccgatttag cagtgtcagc gtattttttc ttctcatcct ggagcgtatt caagatcttc 4620 ccaatacaag aaaattaata aaaaatttat atataggcag cagcaaaaga gccatgttca 4680 aaatagtcat tatgggctca aatagaaaga agacttttaa gttttaatcc agtttatctg 4740 ttgagttctg tgagctactg acctcctgag actggcactg tgtaagtttt agttgcctac 4800 cctagctctt ttctcgtaca attttgccaa taccaagttt caatttgttt ttacaaaaca 4860 ttattcaagc cactagaatt atcaaatatg acgctatagc agagtaaata ctctgaataa 4920 gagaccggta ctagctaact ccaagagatc gttagcagca tcagtccaca aacacttagt 4980 ggcccacaat atatagagag atagaaaagg tagttataac ttgaagcatg tatttaatgc 5040 aaataggcac gaaggcacag gtctaaaata ctacattgtc actgtaagct atacttttaa 5100 aatatttatt ttttttaaag tattttctag tcttttctct ctctgtggaa tggtgaaaga 5160 gagatgccgt gttttgaaag taagatgatg aaatgaattt ttaattcaag aaacattcag 5220 aaacatagga attaaaactt agagaaatga tctaatttcc ctgttcacac aaactttaca 5280 ctttaatctg atgattggat attttatttt agtgaaacat catcttgtta gctaacttta 5340 aaaaatggat gtagaatgat taaaggttgg tatgattttt ttttaatgta tcagtttgaa 5400 cctagaatat tgaattaaaa tgctgtctca gtattttaaa agcaaaaaag gaatggagga 5460 aaattgcatc ttagaccatt tttatatgca gtgtacaatt tgctgggcta gaaatgagat 5520 aaagattatt tatttttgtt catatcttgt acttttctat taaaatcatt ttatgaaatc 5580 caaaaaaaaa aaaaaaaaa 5599

Claims (11)

A composition for diagnosing the severity of an infectious disease, comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.
The composition of claim 1, wherein the Wnt11 protein consists of the amino acid sequence of SEQ ID NO: 1.
The composition of claim 1, wherein the Wnt11 mRNA consists of the amino acid sequence of SEQ ID NO: 2.
The composition of claim 1, wherein the composition further comprises an agent for measuring the expression level of Wnt5a protein or mRNA.
The composition of claim 1, wherein the infectious disease is caused by one or more infections selected from the group consisting of viruses, bacteria and fungi.
According to claim 1, wherein the infectious disease is sepsis, septic shock, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, severe acute respiratory syndrome coronavirus (SARS-CoV) Infectious diseases, Middle East Respiratory Syndrome (MERS), salmonellosis, food poisoning, typhoid, paratyphoid, systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), pneumonia, pulmonary tuberculosis , tuberculosis, cold, influenza, respiratory tract infection, rhinitis, nasopharyngitis, otitis media, bronchitis, lymphadenitis, mumps, lymphadenitis, stomatitis, stomatitis, arthritis, myositis, dermatitis, vasculitis, gingivitis, periodontitis, keratitis, conjunctivitis, wound infection, Peritonitis, hepatitis, osteomyelitis, cellulitis, meningitis, encephalitis, brain abscess, encephalomyelitis, meningitis, osteomyelitis, nephritis, carditis, endocarditis, enteritis, gastritis, esophagitis, duodenitis, colitis, urethritis, cystitis, vaginitis, cervicitis, salpingitis, Infectious erythema, bacterial dysentery, abscesses and ulcers, bacteremia, diarrhea, dysentery, gastroenteritis, gastroenteritis, genitourinary abscess, open wound or wound infection, purulent inflammation, abscess, boil, pyoderma, impetigo, folliculitis, cellulitis, postoperative wound Infection, skin laceration syndrome, skin burn syndrome, thrombotic thrombocytopenia, hemolytic uremic syndrome, renal failure, pyelonephritis, glomerulonephritis, nervous system abscess, otitis media, sinusitis, pharyngitis, tonsillitis, mastoiditis, cellulitis, dental infection, lacrimal cystitis , pleurisy, abdominal abscess, liver abscess, cholecystitis, splenic abscess, pericarditis, myocarditis, placenta, amnioticitis, mastitis, mastitis, puerperal fever, toxic shock syndrome, Lyme disease, gas necrosis, atherosclerosis Sclerosis, mycobacterium avium syndrome (MAC), enterohaemorrhagic E. coli (EHEC) infection, enteropathogenic E. coli (EPEC) infection, enteroinvasive E. coli infection (EIEC), A composition, characterized in that at least one selected from the group consisting of methicillin-resistant Staphylococcus aureus (MRSA) infection, vancomycin-resistant Staphylococcus aureus (VRSA) infection, and listerosis.
A kit for diagnosing the severity of an infectious disease comprising an agent for measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA.
In order to provide information necessary for diagnosing the severity of infectious diseases,
(A) measuring the expression level of Wnt11 (Wnt family member 11) protein or mRNA in a biological sample provided from a patient suspected of an infectious disease;
(b) a method of detecting Wnt11 protein or mRNA, comprising the step of comparing the expression level of the Wnt11 protein or mRNA with a normal person.
The method according to claim 8, wherein the biological sample is at least one selected from the group consisting of blood, plasma, serum, saliva, nasal fluid, sputum, joint capsule fluid, amniotic fluid, ascites, cervical or vaginal secretion, urine, and cerebrospinal fluid. How to.
The method according to claim 8, wherein the expression level of Wnt5a protein or mRNA is further measured in step (a), and the lower the Wnt11/Wnt5a value compared to the normal in step (b), the higher the disease severity is determined. how to do it
Wnt11 (Wnt family member 11) protein, a vector comprising a polynucleotide encoding the Wnt11 protein, or a pharmaceutical composition for preventing or treating an infectious disease comprising a cell comprising the vector as an active ingredient.

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