WO2016167573A1 - Composition de biomarqueur pour le diagnostic de la maladie de still, trousse de diagnostic et procédé de diagnostic - Google Patents

Composition de biomarqueur pour le diagnostic de la maladie de still, trousse de diagnostic et procédé de diagnostic Download PDF

Info

Publication number
WO2016167573A1
WO2016167573A1 PCT/KR2016/003898 KR2016003898W WO2016167573A1 WO 2016167573 A1 WO2016167573 A1 WO 2016167573A1 KR 2016003898 W KR2016003898 W KR 2016003898W WO 2016167573 A1 WO2016167573 A1 WO 2016167573A1
Authority
WO
WIPO (PCT)
Prior art keywords
disease
still
cd11b
sample
lymphocytes
Prior art date
Application number
PCT/KR2016/003898
Other languages
English (en)
Korean (ko)
Inventor
손성향
김현아
Original Assignee
아주대학교산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020160044090A external-priority patent/KR101806681B1/ko
Application filed by 아주대학교산학협력단 filed Critical 아주대학교산학협력단
Priority to EP16780296.6A priority Critical patent/EP3285071B1/fr
Publication of WO2016167573A1 publication Critical patent/WO2016167573A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

Definitions

  • the present invention relates to a composition for diagnosing Still's disease, a diagnostic kit, and a method for providing information necessary for diagnosis.
  • Still's disease is a systemic form of juvenile rheumatoid arthritis that has not yet been identified since it was first described and develops in both children and adults.
  • Adult Still's disease, which occurs in adults, has been reported all over the world since it was reported by Bywaters in 1971, and is known as a rare disease or a cause of unknown fever.
  • Still's disease is mainly based on clinical findings, including high fever, arthralgia or arthritis, characteristic skin lesions, lymphadenopathy, hepatomegaly, splenomegaly, meningitis and sore throat, leukocytosis, increased blood infiltration rate, Diagnosis is based on laboratory findings such as antibody negative and rheumatoid factor negative.
  • HLAs human leucocyte antigens
  • DAMPs damage-associated molecular patterns
  • MAMPs microbe-associated molecular patterns
  • an object of the present invention is to provide a biomarker composition capable of diagnosing Still's disease.
  • Another object of the present invention is to provide a kit for diagnosing Still's disease.
  • Another object of the present invention is to provide a method for providing information necessary for diagnosing Still's disease.
  • the present invention provides a biomarker composition for diagnosing Stillse disease comprising at least one protein selected from CD11b or CD32.
  • the present invention provides a stye disease diagnostic kit comprising an antibody that specifically binds to CD11b or CD32 cell surface antigen.
  • the present invention comprises the steps of measuring the expression level of one or more proteins selected from CD11b or CD32 from the sample; Comparing the expression level with the expression level of a normal control sample; And diagnosing Still's disease when at least one expression level selected from CD11b or CD32 of the sample is higher than that of the normal control sample.
  • the present invention comprises the steps of measuring the frequency ratio of granulocytes to lymphocytes from the sample; Comparing the frequency ratio with the frequency ratio of a normal control sample; And it provides a method for providing information necessary for diagnosing Still's disease, if the frequency ratio of granulocytes to lymphocytes of the sample is higher than the normal control sample.
  • the frequency of CD11b-expressing cells and CD32-expressing cells is higher, and lymphocytes
  • the frequency ratio of the granulocytes to the other also shows a higher difference, which is why still's disease comprising the antibody specifically binding to the CD11b or CD32 cell surface antigen, the diagnostic biomarker composition comprising one or more proteins selected from CD11b or CD32
  • PBMC peripheral blood mononuclear cells
  • FIG. 2 is an image showing the frequency of living cells (P1%), the frequency of cells expressing CD11b and the frequency of cells expressing CD32 in PBMC,
  • 3 is a table showing the ratio of granulocytes, lymphocytes and lymphocytes to lymphocytes in PBMC,
  • Figure 4 is an image showing the ratio of granulocytes, lymphocytes in PBMC.
  • the inventor of the present invention while studying a biomarker for diagnosing Still's disease, the frequency of CD11b-expressing cells, the frequency of CD32-expressing cells and the frequency of granulocytes for lymphocytes in normal people and patients with Still's disease. The difference in the ratio was found to complete the present invention.
  • the frequency means the number of cells expressing the CD11b, the number of cells expressing CD32, and the number of frequencies occupied by granulocytes or lymphocytes.
  • the present invention provides a biomarker composition for diagnosing Still's disease, which comprises at least one protein selected from CD11b or CD32.
  • the frequency of CD11b-expressing cells and CD32-expressing cells in the whole peripheral blood mononuclear cells (PBMC) of the Still's disease patients, respectively, of the PBMCs of normal or rheumatic patients Expression was higher than the frequency of cells expressing CD11b or CD32.
  • the proportion of granulocytes in PBMCs of patients with Stillse disease was higher than that of granulocytes in PBMCs of normal or rheumatic patients, and the proportion of lymphocytes was lower.
  • the present invention provides a kit for diagnosing Still's disease, which comprises an antibody that specifically binds to CD11b or CD32 cell surface antigen.
  • the antibody is a polyclonal antibody or a monoclonal antibody.
  • the polyclonal antibody can be prepared by injecting a CD11b or CD32 protein or fragment thereof into an external host with an immunogen according to methods known to those skilled in the art.
  • External hosts include mammals such as mice, rats, sheep or rabbits.
  • Immunogens are injected by intramuscular, intraperitoneal or subcutaneous injection and are usually administered with an adjuvant to increase antigenicity. Serum is periodically collected from external hosts to collect serum that shows improved titers and specificity for antigen or to separate and purify antibodies therefrom.
  • the monoclonal antibodies can be prepared by the technology of generating immortalized cell lines by fusion known to those skilled in the art.
  • the protein expressed by the gene is immunized in mice by immunizing mice or by synthesizing peptides and binding to bovine serum albumin.
  • Antigen-producing B lymphocytes isolated from mice are fused with myeloma of human or mouse to produce immortalized hybridomas, indirect enzyme-linked immunosorbent assays with the hybridoma cells.
  • Monoclonal antibodies can be prepared by confirming the production of monoclonal antibodies using linked immunoabsorbent assays (ELISA), selecting positive clones, incubating them, and then separating or purifying the antibodies or injecting them into the abdominal cavity of rats. have.
  • ELISA linked immunoabsorbent assays
  • the monoclonal antibody is generally quantified by color reaction using a secondary antibody to which an enzyme such as alkaline phosphatase (AP) or horseradish peroxidase (HRP) is bound, and a substrate thereof.
  • an enzyme such as alkaline phosphatase (AP) or horseradish peroxidase (HRP) is bound
  • the antibody may be directly quantitated using a combination of an AP or HRP enzyme or the like with a monoclonal antibody directed against the protein.
  • antibodies with various fluorescent labels including fluorescein isothiocyanat (FITC), phycoerythrin (PE), and the like can also be used.
  • the reaction between the protein and the antibody is Western blot, immunoprecipitation (IP), enzyme-linked immunoabsorbent assays (ELISA), immunohistochemistry (IHC) and flow cytometry (Flow) Protein identification experiments, such as cytometry analysis (FACS), can be confirmed, but are not limited thereto.
  • Kits comprising such antibodies typically include lyophilized antibodies, buffers, stabilizers, inactive proteins, and the like, wherein the antibodies are prepared by radionuclides, fluorescors, enzymes, and the like. Can be labeled.
  • the present invention comprises the steps of measuring the expression level of at least one protein selected from CD11b or CD32; Comparing the expression level with the expression level of a normal control sample; And diagnosing Still's disease when at least one expression level selected from CD11b or CD32 of the sample is higher than that of the normal control sample.
  • the present invention comprises the steps of measuring the frequency ratio of granulocytes to lymphocytes from the sample; Comparing the frequency ratio with the frequency ratio of a normal control sample; And it provides a method for providing information necessary for diagnosing Still's disease, if the frequency ratio of granulocytes to lymphocytes of the sample is higher than the normal control sample.
  • the frequency ratio of granulocytes to lymphocytes can be diagnosed as Still's disease when 3.0 to 50.0.
  • the sample uses blood, and more preferably, it is preferable to use peripheral blood mononuclear cells (PBMC) isolated from blood, but is not limited thereto.
  • PBMC peripheral blood mononuclear cells
  • PBMCs Human PBMCs were isolated from human blood from healthy volunteer donors using Ammonium-Chloride-Potassium (ACK) Lysis Buffer. Ethylenediaminetetraacetic acid (EDTA) was added to 2 mL of blood in a tube treated with 15 inversions. It was then placed in 40 mL ACK lysis buffer and left at room temperature for 5 minutes. White blood cells (WBC) were collected by centrifugation at 300 xg for 5 minutes, the supernatant was discarded, and the precipitated cells were mixed in 5 ml PBS. PBMC was prepared by repeating the centrifugation twice for 5 minutes at a speed of 300 xg at 4 ° C.
  • WBC White blood cells
  • Example 2 peripheral blood mononuclear cells, PBMC )in CD11b Or measuring the frequency of cells expressing CD32
  • red blood cells are removed from each blood, lymphocytes are collected and reacted with anti-CD11b and anti-CD32 antibodies with different fluorescence, and then the number of cells with antibodies is counted using flow cytometry (FACS). And the frequency which these cells occupy in the whole cell was analyzed.
  • FACS flow cytometry
  • the frequency (P1%) of living cells in the total cells did not have a big difference as a whole.
  • Example 3 peripheral blood mononuclear cells, PBMC Frequency of granulocytes and lymphocytes
  • Example 2 In the same manner as in Example 2, the frequency of granulocytes and lymphocytes was measured in PBMCs obtained from normal (19 patients), rheumatoid patients (RA) (19), and Still's disease patients (18), and the ratio of granulocytes to lymphocytes was measured. Confirmed.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne une composition de biomarqueur pour le diagnostic de la maladie de Still, une trousse de diagnostic et un procédé de diagnostic, et plus spécifiquement, la présente invention permet de diagnostiquer avec précision la maladie de Still chez les jeunes enfants et les adultes à l'aide du taux d'expression de CD11b ou CD32 et du rapport de fréquence des granulocytes aux lymphocytes en tant qu'index.
PCT/KR2016/003898 2015-04-15 2016-04-14 Composition de biomarqueur pour le diagnostic de la maladie de still, trousse de diagnostic et procédé de diagnostic WO2016167573A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP16780296.6A EP3285071B1 (fr) 2015-04-15 2016-04-14 Utilisation d'une composition de biomarqueur ou trousse de diagnostic pour le diagnostic de la maladie de still et procédé de diagnostic

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2015-0053231 2015-04-15
KR20150053231 2015-04-15
KR10-2016-0044090 2016-04-11
KR1020160044090A KR101806681B1 (ko) 2015-04-15 2016-04-11 스틸씨병 진단용 바이오마커 조성물, 진단 키트 및 진단 방법

Publications (1)

Publication Number Publication Date
WO2016167573A1 true WO2016167573A1 (fr) 2016-10-20

Family

ID=57125937

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2016/003898 WO2016167573A1 (fr) 2015-04-15 2016-04-14 Composition de biomarqueur pour le diagnostic de la maladie de still, trousse de diagnostic et procédé de diagnostic

Country Status (1)

Country Link
WO (1) WO2016167573A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1111049A1 (fr) * 1998-09-03 2001-06-27 Asahi Kasei Kabushiki Kaisha Nouvelle proteine de recepteur et procede servant a diagnostiquer des maladies inflammatoires au moyen de cette proteine
KR20140093901A (ko) * 2013-01-18 2014-07-29 서울대학교산학협력단 류마티스 관절염 진단용 바이오마커

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1111049A1 (fr) * 1998-09-03 2001-06-27 Asahi Kasei Kabushiki Kaisha Nouvelle proteine de recepteur et procede servant a diagnostiquer des maladies inflammatoires au moyen de cette proteine
KR20140093901A (ko) * 2013-01-18 2014-07-29 서울대학교산학협력단 류마티스 관절염 진단용 바이오마커

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JUNG, YOON SUK ET AL.: "A Case of Adult-onset Still's Disease Combined with Sjogren's Syndrome", THE KOREAN JOURNAL OF INTERNAL MEDICINE, vol. 77, no. 1, 2009, pages S240 - S244, XP009507331 *
SEUNG, O. P. ET AL.: "Adult-onset Still's Disease: a Case Report", OMAN MEDICAL JOURNAL, vol. 26, no. 5, 2011, pages 1 - 3, XP055497994 *
YAMAGUCHI, M. ET AL.: "Preliminary Criteria for Classification of Adult Still's Disease", THE JOURNAL OF RHEUMATOLOGY, vol. 19, no. 3, March 1992 (1992-03-01), pages 424 - 430, XP009507351 *

Similar Documents

Publication Publication Date Title
Hess Routine Epstein-Barr virus diagnostics from the laboratory perspective: still challenging after 35 years
KR101806681B1 (ko) 스틸씨병 진단용 바이오마커 조성물, 진단 키트 및 진단 방법
AU2005259346B2 (en) Detection of tuberculosis and infection by mycobacterium tuberculosis using HBHA
Pfrepper et al. Seroreactivity to and avidity for recombinant antigens in toxoplasmosis
Abd-Alla et al. Differentiation of pathogenic Entamoeba histolytica infections from nonpathogenic infections by detection of galactose-inhibitable adherence protein antigen in sera and feces
Origgi et al. Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni])
Heide et al. Broadly directed SARS-CoV-2-specific CD4+ T cell response includes frequently detected peptide specificities within the membrane and nucleoprotein in patients with acute and resolved COVID-19
CN1656378A (zh) 诊断病毒感染的方法
Saliki et al. Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detection of morbillivirus antibody in marine mammal sera
Demaret et al. Severe SARS‐CoV‐2 patients develop a higher specific T‐cell response
Ribeiro et al. Investigation of the prevalence of antibodies against neurotropic polyomaviruses BK, JC and SV40 in sera from patients affected by multiple sclerosis
Brunner et al. Use of serum immune complexes in a new test that accurately confirms early Lyme disease and active infection with Borrelia burgdorferi
CN111004324A (zh) 一种钙卫蛋白单克隆抗体及其应用
JPH11237381A (ja) ヒトまたは動物の移植用組織をそのレシピエントに 適合させる方法及びヒトまたは動物において 腫瘍抗原を検出する方法
Diaz et al. The IgM anti-desmoglein 1 response distinguishes Brazilian pemphigus foliaceus (fogo selvagem) from other forms of pemphigus
Saxinger et al. Possible risk to recipients of blood from donors carrying serum markers of human T-cell leukaemia virus
AU2012298229A1 (en) Improved vaccine diagnostics
Nobuo et al. Enzyme-linked immunosorbent assay for IgG and IgM antibodies against human parvovirus B19: use of monoclonal antibodies and viral antigen propagated in vitro
US10634676B2 (en) Method and kit for simultaneously detecting human parvovirus B19 antigen and antibody
WO2012144784A2 (fr) Anticorps monoclonaux qui reconnaissent spécifiquement la carboxylestérase 1 de foie humain, lignées cellulaires d'hybridome qui produisent les anticorps monoclonaux et leurs utilisations
WO2016167573A1 (fr) Composition de biomarqueur pour le diagnostic de la maladie de still, trousse de diagnostic et procédé de diagnostic
EP1240519B1 (fr) Compositions et procedes pour la detection du treponema pallidum
Zhang et al. Autoantibody responses in Chinese hepatocellular carcinoma
Nollens et al. Cross-reactivity between immunoglobulin G antibodies of whales and dolphins correlates with evolutionary distance
JPH0328766A (ja) Ｈｉｖ抗体の検出方法およびそれに用いるアッセイキット

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16780296

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE