MD1009Z - Method for determining the susceptibility of a person to the development of non-Hodgkin lymphoma - Google Patents
Method for determining the susceptibility of a person to the development of non-Hodgkin lymphoma Download PDFInfo
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- MD1009Z MD1009Z MDS20150029A MDS20150029A MD1009Z MD 1009 Z MD1009 Z MD 1009Z MD S20150029 A MDS20150029 A MD S20150029A MD S20150029 A MDS20150029 A MD S20150029A MD 1009 Z MD1009 Z MD 1009Z
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Abstract
Изобретение относится к медицине, в частности к онкохематологии и может быть использовано для определения предрасположенности человека к развитию лимфомы нон-Ходжкин.Согласно изобретению, метод состоит в том, что у лиц, у которых определяют генотип Arg/Arg кодона гена XRCC1, дополнительно определяют и соотношение иммуноглобулинов IgM и IgG и в случае, когда соотношение составляет 0,03…0,24, констатируют предрасположенность человека к развитию лимфомы нон-Ходжкин.The invention relates to medicine, in particular to oncohematology and can be used to determine a person’s predisposition to the development of non-Hodgkin lymphoma. According to the invention, the method consists in the fact that in individuals who determine the Arg / Arg genotype of the codon of the XRCC1 gene, they additionally determine the ratio of IgM and IgG immunoglobulins and in the case when the ratio is 0.03 ... 0.24, a person’s predisposition to the development of non-Hodgkin lymphoma is ascertained.
Description
Invenţia se referă la medicină, în special la oncohematologie şi poate fi utilizată pentru determinarea predispunerii persoanei la dezvoltarea limfomului non-Hodgkin. The invention relates to medicine, in particular to oncohematology and can be used to determine a person's predisposition to developing non-Hodgkin lymphoma.
Limfomul non-Hodgkin (LNH) este o formă de cancer care se dezvoltă în ţesutul limfatic - situat în diverse părţi ale organismului: ganglioni limfatici, splină, timus, vegetaţii adenoide, amigdale şi maduva osoasă. Non-Hodgkin lymphoma (NHL) is a form of cancer that develops in the lymphatic tissue - located in various parts of the body: lymph nodes, spleen, thymus, adenoids, tonsils and bone marrow.
Sistemul limfatic este o parte importantă a sistemului imun, joacă un rol esenţial în apărarea organismului contra infecţiilor. Limfomul non-Hodgkin işi are originea în ţesutul limfatic, dar se poate răspândi şi în alte organe. The lymphatic system is an important part of the immune system, playing a key role in defending the body against infections. Non-Hodgkin lymphoma originates in the lymph tissue, but can also spread to other organs.
Limfomul non-Hodgkin se împarte în două categorii: Non-Hodgkin lymphoma is divided into two categories:
Limfom non-Hodgkin agresiv caracterizat prin rapiditatea cu care se divid celulele neoplazice din ganglionii limfatici, necesitând tratament imediat. Fară un tratament adecvat, speranţa medie de viaţă la pacienţii cu limfom non-Hodgkin agresiv este de la şase luni la 2 ani. Pacienţii diagnosticaţi şi trataţi în stadiile incipiente ale bolii au o şansă mai mare de remisie completă cu durata de câţiva ani şi o probabilitate mai mică de recurenţă a bolii. Aggressive non-Hodgkin lymphoma characterized by the rapid division of neoplastic cells in the lymph nodes, requiring immediate treatment. Without appropriate treatment, the average life expectancy for patients with aggressive non-Hodgkin lymphoma is six months to 2 years. Patients diagnosed and treated in the early stages of the disease have a greater chance of complete remission lasting several years and a lower probability of recurrence of the disease.
Limfomul non-Hodgkin indolent este caracterizat prin multiplicarea şi răspândirea mai lentă în organism a celulelor neoplazice, ceea ce face diagnosticarea mai dificilă. Pacienţii pot trăi în medie 10 ani cu această boală; un pacient cu limfom non-Hodgkin indolent poate primi tratament de 5-6 ori pe parcursul vieţii, pentru a încetini progresia bolii. Tratamentele standard nu pot vindeca această formă de limfom. Indolent non-Hodgkin lymphoma is characterized by the slower multiplication and spread of neoplastic cells in the body, which makes diagnosis more difficult. Patients can live an average of 10 years with this disease; a patient with indolent non-Hodgkin lymphoma may receive treatment 5-6 times during their lifetime to slow the progression of the disease. Standard treatments cannot cure this form of lymphoma.
Este cunoscută o metodă de determinare a predispunerii persoanei la dezvoltarea limfomului non-Hodgkin, care constă în aceea că se cercetează genotipul 399 al codonului genei XRCC1 şi în cazul depistării genotipului Gln/Gln se determină o predispunere a persoanei la dezvoltarea limfomului non-Hodgkin agresiv, iar în cazul când se determină genotipul Arg/Arg se determină un risc minim de predispunere a persoanei la dezvoltarea limfomului non-Hodgkin agresiv [1]. A method is known to determine a person's predisposition to developing non-Hodgkin lymphoma, which consists of researching the genotype 399 of the XRCC1 gene codon and in case of detecting the Gln/Gln genotype, a predisposition of the person to developing aggressive non-Hodgkin lymphoma is determined, and in case of determining the Arg/Arg genotype, a minimal risk of predisposition of the person to developing aggressive non-Hodgkin lymphoma is determined [1].
Dezavantajul metodei constă în aceea că nu este certă în determinarea predispunerii persoanei la dezvoltarea limfomului non-Hodgkin, întrucât chiar şi la persoanele la care se determină genotipul Arg/Arg, care este un indicator minim de dezvoltare a maladiei, şi la ele se pot dezvolta limfoame non-Hodgkin. Într-un studiu efectuat pe un lot de 26 de persoane la care s-a determinat genotipul Arg/Arg, la 7 din ele s-a dezvoltat limfomul non-Hodgkin. The disadvantage of the method is that it is not certain in determining a person's predisposition to developing non-Hodgkin lymphoma, since even people with the Arg/Arg genotype, which is a minimal indicator of the development of the disease, can still develop non-Hodgkin lymphoma. In a study conducted on a group of 26 people with the Arg/Arg genotype, 7 of them developed non-Hodgkin lymphoma.
Problema pe care o rezolvă invenţia constă în extinderea gamei de metode destinate determinării predispunerii persoanelor la dezvoltarea limfomului non-Hodgkin prin elaborarea unei metode, care ar permite cert determinarea predispunerii la dezvoltarea limfomului non-Hodgkin. The problem solved by the invention consists in expanding the range of methods intended to determine the predisposition of individuals to the development of non-Hodgkin lymphoma by developing a method that would certainly allow determining the predisposition to the development of non-Hodgkin lymphoma.
Esenţa metodei de determinare a predispunerii persoanei la dezvoltarea limfomului non-Hodgkin constă în aceea că la persoanele la care se determină genotipul Arg/Arg al codonului genei XRCC1 suplimentar se determină şi raportul imunoglobulinelor IgM şi IgG şi în cazul în care raportul constituie 0,03…0,24, se constată predispunerea persoanei la dezvoltarea limfomului non-Hodgkin. The essence of the method for determining a person's predisposition to developing non-Hodgkin lymphoma is that in people in whom the Arg/Arg genotype of the XRCC1 gene codon is determined, the ratio of IgM and IgG immunoglobulins is additionally determined and if the ratio is 0.03…0.24, the person's predisposition to developing non-Hodgkin lymphoma is determined.
Rezultatul invenţiei constă în aceea că datorită metodei revendicate se poate de determinat cert predispunerea persoanelor la dezvoltarea limfomului non-Hodgkin. The result of the invention is that thanks to the claimed method, the predisposition of individuals to the development of non-Hodgkin lymphoma can be determined with certainty.
Datorită metodei elaborate se poate cert de determinat predispunerea la dezvoltarea limfomului non-Hodgkin chiar şi la persoanele la care se determină genotipul Arg/Arg, care este un indicator minim de dezvoltare a maladiei non-Hodgkin la pacienţii cu genotipul Arg/Arg al genei XRCC1. Thanks to the developed method, it is possible to reliably determine the predisposition to the development of non-Hodgkin lymphoma even in people with the Arg/Arg genotype, which is a minimal indicator of the development of non-Hodgkin disease in patients with the Arg/Arg genotype of the XRCC1 gene.
Metoda se efectuează în felul următor. The method is performed as follows.
Din patul vascular se colectează 10,0 mL de sânge, se evidenţiază ADN-ul, apoi se efectuează genotiparea polimorfismului Arg399Gln al genei XRCC1 care include reacţia de polimerizare în lanţ efectuată cu ajutorul termociclerului automat „Eppendorf” ce constă din 31 de cicluri, apoi se efectuează analiza electroforetică a reacţiei de polimerizare în lanţ pe un gel agarizat de 1,5% cu obţinerea fragmentelor genei XRCC1, se efectuează hidroliza cu endonucleaza de restricţie MspI, timp de 3 ore la 37 °C, după care se face analiza electroforetică pe mediu gelifiant agarizat de 1,5% a endonucleazei de restricţie MspI şi dacă se observă o fâşie, atunci se determină genotipul Gln/Gln, 2 fâşii pe banda electroforetică vorbesc despre genotipul Arg/Arg, iar 3 fâşii vorbesc despre genotipul Arg/Gln. La persoanele cu genotipul Arg/Arg se determină cantitatea de IgM şi IgG, după care se calculează raportul dintre ele şi în cazul în care raportul constituie 0,03…0,24, se determină o predispunere a persoanei la dezvoltarea limfomului non-Hodgkin. 10.0 mL of blood is collected from the vascular bed, the DNA is highlighted, then the genotyping of the Arg399Gln polymorphism of the XRCC1 gene is performed, which includes the polymerization chain reaction performed using the "Eppendorf" automatic thermocycler consisting of 31 cycles, then the electrophoretic analysis of the polymerization chain reaction is performed on a 1.5% agarized gel to obtain fragments of the XRCC1 gene, hydrolysis is performed with the restriction endonuclease MspI, for 3 hours at 37 °C, after which the electrophoretic analysis is performed on a 1.5% agarized gelling medium of the restriction endonuclease MspI and if a band is observed, then the Gln/Gln genotype is determined, 2 bands on the electrophoretic strip speak of the Arg/Arg genotype, and 3 bands speak of the Arg/Gln genotype. In people with the Arg/Arg genotype, the amount of IgM and IgG is determined, after which the ratio between them is calculated and if the ratio is 0.03…0.24, the person's predisposition to developing non-Hodgkin lymphoma is determined.
Ca rezultat al cercetărilor efectuate s-a constatat că odată cu creşterea IgG şi micşorarea IgM în patul sanguin creşte probabilitatea de dezvoltare la persoanele cu genotipul Arg/Arg a maladiei limfomului non-Hodgkin, la fel s-a determinat şi raportul care cert indică la apariţia riscului de dezvoltare a maladiei menţionate. Analizele au fost efectuate pe un lot de 26 de pacienţi, dintre care la 7 s-a depistat genotipul Arg/Arg al genei XRCC1. La toţi cei 7 pacienţi au fost depistate rapoarte IgM faţă de IgG în intervalul 0,03…0,24, iar la ceilalţi 19 pacienţi raportul a constituit mai mult de 0,25. As a result of the research, it was found that with the increase in IgG and the decrease in IgM in the bloodstream, the probability of developing non-Hodgkin lymphoma in people with the Arg/Arg genotype increases, and the ratio that definitely indicates the risk of developing the disease was also determined. The analyzes were performed on a group of 26 patients, of which 7 had the Arg/Arg genotype of the XRCC1 gene. In all 7 patients, IgM to IgG ratios were detected in the range of 0.03…0.24, and in the other 19 patients the ratio was more than 0.25.
În anul 1970, prin consens între specialişti, Organizaţia Mondială a Sănătăţii a stabilit ca substanţele cu proprietăţi de anticorp să fie grupate în categoria imunoglobulinelor, pornind de la faptul că toate substanţele din acest grup au funcţie imună şi sunt cuprinse în fracţia globulinică a serului. In 1970, by consensus among specialists, the World Health Organization established that substances with antibody properties should be grouped in the immunoglobulin category, based on the fact that all substances in this group have an immune function and are contained in the globulin fraction of serum.
Anticorpii sunt imunoglobuline care se sintetizează în organism după pătrunderea unui antigen şi au proprietatea de a se cupla specific cu antigenul inductor şi de a-i anihila acţiunea nocivă, anticorpii sau Ig sunt produsul final al reacţiei imune (RIU). Antibodies are immunoglobulins that are synthesized in the body after the entry of an antigen and have the property of specifically coupling with the inducing antigen and annihilating its harmful action. Antibodies or Ig are the final product of the immune reaction (RIU).
Exemplu Example
Pacienta A., 51 ani, s-a adresat la centrul Consultativ Diagnostic al IOM la un control profilactic. Bolnava în anamneză suferă de ulcer gastric de 8 ani, s-a depistat infectarea cu Helicobacter pylori netratată, cu 3 ani în urmă a suferit o mastectomie pe stânga, după care a suportat un curs de chimioterapie. Bolnava în cadrul IOM a fost supusă examenului clinic şi paraclinic, ganglionii limfatici regionali nu se palpau, iar în cadrul examenului paraclinic s-a efectuat radiografia cutiei toracice, ultrasonografia organelor interne, analiza generală a sângelui, toate datele din investigaţii au fost în limitele fiziologice, complicaţii legate de intervenţia suportată după mastectomie nu s-au depistat, apoi bolnava a fost examinată la prezenţa genotipării polimorfismului Arg399Gln al genei XRCC1, s-a determinat prezenţa genotipului Arg/Arg, după care s-a determinat şi cantitatea de IgM care a constituit 3,0 g/L şi cantitatea de IgG - 25,0 g/L, raportul acestor imunoglobuline a constituit 0,12. Acest raport indică o predispunere a persoanei cu prezenţa genotipului Arg/Arg la dezvoltarea limfomului non-Hodgkin. Bolnava a refuzat să trateze infecţia de Helicobacter pylori, dar şi să urmeze o cură de tratament de susţinere şi peste 3,5 luni s-a adresat la centrul Consultativ Diagnostic al IOM cu acuze la apariţia unui ganglion limfatic cervical pe dreapta, indolor, bolnava a fost investigată clinic şi paraclinic, în cadrul investigărilor s-au efectuat: Patient A., 51 years old, went to the IOM Diagnostic Consultative Center for a prophylactic check-up. The patient has a history of gastric ulcer for 8 years, was diagnosed with untreated Helicobacter pylori infection, and 3 years ago underwent a left mastectomy, after which she underwent a course of chemotherapy. The patient at the IOM underwent clinical and laboratory examination, regional lymph nodes were not palpable, and during the laboratory examination, a chest X-ray, ultrasound of internal organs, and a general blood test were performed, all the data from the investigations were within physiological limits, complications related to the intervention after the mastectomy were not detected, then the patient was examined for the presence of the Arg399Gln polymorphism genotyping of the XRCC1 gene, the presence of the Arg/Arg genotype was determined, after which the amount of IgM was also determined, which was 3.0 g/L and the amount of IgG - 25.0 g/L, the ratio of these immunoglobulins was 0.12. This ratio indicates a predisposition of the person with the presence of the Arg/Arg genotype to the development of non-Hodgkin lymphoma. The patient refused to treat the Helicobacter pylori infection, but also to follow a supportive treatment course and after 3.5 months she went to the IOM Diagnostic Consultative Center with allegations of the appearance of a painless cervical lymph node on the right. The patient was investigated clinically and paraclinically, during the investigations the following were performed:
Analiza generală a sângelui date de referinţă Hemoglobina - 126 g/L 137…175 g/L Eritrocite - 4,3 x1012/L 4,63…6,1x 109 g/L Hematocrit - 44,3% 40…51% Reticulocite - 1,84% 0,5…1,7% Trombocite - 494,5 x109/L 163…380 x109/L Leucocite - 11,54 x109/L 4,23… 9,07x109/L Nesegmentate - 2% 0… 2,5% Segmentate - 54% 38…78% Eozinofile - 3% 0,8…7% Bazofile - 0,5% 0,1…1,2% Limfocite - 34% 19…37% Monocite - 5% 5,3…12,2% VSH - 20 mm/oră 2…10 mm/oră Volumul eritrocitar mediu - 84,9 fL 79…92,2 fL Hemoglobina eritrocitară medie - 29,5 pg 25,7…32,2 pg Concentraţia medie a hemoglobinei - 34,8 g/dL 32,3… 36,5 g/L Gradul de anizocitoză - 12,8% 11,6…14,4% Volumul mediu al trombocitelor - 10,6 fL 9,4…12,4 fLGeneral blood analysis reference data Hemoglobin - 126 g/L 137…175 g/L Erythrocytes - 4.3 x1012/L 4.63…6.1x 109 g/L Hematocrit - 44.3% 40…51% Reticulocytes - 1.84% 0.5…1.7% Platelets - 494.5 x109/L 163…380 x109/L Leukocytes - 11.54 x109/L 4.23… 9.07x109/L Non-segmented - 2% 0… 2.5% Segmented - 54% 38…78% Eosinophils - 3% 0.8…7% Basophils - 0.5% 0.1…1.2% Lymphocytes - 34% 19…37% Monocytes - 5% 5.3…12.2% ESR - 20 mm/hour 2…10 mm/hour Mean erythrocyte volume - 84.9 fL 79…92.2 fL Mean erythrocyte hemoglobin - 29.5 pg 25.7…32.2 pg Mean hemoglobin concentration - 34.8 g/dL 32.3… 36.5 g/L Degree of anisocytosis - 12.8% 11.6…14.4% Mean platelet volume - 10.6 fL 9.4…12.4 fL
Analiza biochimică a sângelui datele de referinţă Proteina generală - 73,0 mmol/L 62,0… 80,0 mmol/L Albumina - 47,73 g/L 34…48 g/L Ureea - 5,76 mmol/L 1,33…11,9 mmol/L Creatinina - 78,39 µmol/L 61,9…106,2 µmol/L Bilirubina totală - 6,16 µmol/L 0…17 µmol/L Bilirubina conjugată - 1,95 µmol/L 0…3,4 µmol/L Bilirubina liberă - 4,207 µmol/L 13,6 µmol/L Glucoza - 5,23 mmol/L 4,1…5,9 mmol/L ALAT - 60,52 U/L 5…41 U/L ASAT - 23,62 U/L 10…50 U/L Amilaza - 53,89 U/L 13…100 U/L Lipaza - 32,86 U/L 13…60 U/L Fosfataza alcalină - 78,58 U/L 40…130 U/L GGTP - 48,15 U/L 10…71 U/L C - proteina reactivă (PCR) - 9,52 mg/L 0,1…5 mg/L Biochemical blood analysis reference data General protein - 73.0 mmol/L 62.0...80.0 mmol/L Albumin - 47.73 g/L 34...48 g/L Urea - 5.76 mmol/L 1.33...11.9 mmol/L Creatinine - 78.39 µmol/L 61.9...106.2 µmol/L Total bilirubin - 6.16 µmol/L 0...17 µmol/L Conjugated bilirubin - 1.95 µmol/L 0...3.4 µmol/L Free bilirubin - 4.207 µmol/L 13.6 µmol/L Glucose - 5.23 mmol/L 4.1...5.9 mmol/L ALT - 60.52 U/L 5...41 U/L SAT - 23.62 U/L 10...50 U/L Amylase - 53.89 U/L 13…100 U/L Lipase - 32.86 U/L 13…60 U/L Alkaline phosphatase - 78.58 U/L 40…130 U/L GGTP - 48.15 U/L 10…71 U/L C - reactive protein (PCR) - 9.52 mg/L 0.1…5 mg/L
A fost efectuată biopsia ganglionului limfatic: Limfom non-Hodgkin varianta limfoblastică şi exulceraţie profundă. Ganglioni limfatici regionali intacţi. Lymph node biopsy was performed: Lymphoblastic variant non-Hodgkin lymphoma and deep exulceration. Intact regional lymph nodes.
Datele obţinute de la examenul clinic şi paraclinic vorbesc despre debutul maladiei non-Hodgkin, ceea ce confirmă datele obţinute cu 3,5 luni în urmă, persoanei i s-a prescris un tratament specific, complicaţii după tratamentul specific nu s-au înregistrat. The data obtained from the clinical and paraclinical examination speak of the onset of non-Hodgkin's disease, which confirms the data obtained 3.5 months ago, the person was prescribed a specific treatment, and no complications after the specific treatment were recorded.
1. RU 2373862 C1 2009.11.27 1. RU 2373862 C1 2009.11.27
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| MD1367C2 (en) * | 1992-11-13 | 2000-11-30 | Idec Pharmaceuticals Corporation | Methods of B-cells lymphoma treatment, anti-CD20 antibodies, hybridoma |
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