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العنوان
Detection of STAT4 in Multiple Sclerosis patients by polymerase chain reaction and flow cytometry/
المؤلف
Salah El-Din, Mai Mohamed,
هيئة الاعداد
باحث / مي محمد صلاح الدين سيد
مشرف / احمد كامل مصطفى
مناقش / هبه احمد
مناقش / احمد عبدالسميع
الموضوع
Clinical Pathology
تاريخ النشر
2024.
عدد الصفحات
152 P. :
اللغة
الإنجليزية
الدرجة
الدكتوراه
التخصص
الطب
الناشر
تاريخ الإجازة
19/12/2023
مكان الإجازة
جامعة أسيوط - كلية الطب - الباثولوجيا الاكلينيكية
الفهرس
Only 14 pages are availabe for public view

from 177

from 177

Abstract

Multiple sclerosis (MS) is chronic autoimmune diseases with strong genetic and environmental components. MS can be triggered by various environmental components, such as exposure to ultraviolet light, drugs, chemicals, and viral infections. The course of MS disease is categorized based on how clinical manifestations develop over time and on the severity of the disease.
The human signal transducer and activator of transcription (STAT) genes have been identified in three chromosomal clusters: STAT1 and STAT4 on human chromosome 2 (q12-33); STAT2 and STAT6 on chromosome 12 (q13-14); and STAT3, STAT5a, and 5b on chromosome 17 (q11.2-22). This gene encodes a transcription factor that can be activated by interleukin (IL)-12 and IL-23 and plays a role in the signaling via type-1 interferon (IFN I) receptor.
STAT4 is essential for IL-12 signaling and induces interferon-gamma (IFNγ) production. Signal transmission from the interferons involves STAT1 and STAT4. The extensive involvement of type I and type II interferons in the pathogenesis of MS and SLE made STAT4 an obvious candidate region for genetic predisposition to these autoimmune diseases. Moreover, the requirement of STAT4 in IL-23-induced IL-17 production has been suggested.
Aim of study:
Determination the level of STAT4 gene expression in different cases of multiple sclerosis. As well as, determination the association of MS active disease with both different STAT4 genotypes detected be PCR and various STAT4 proteins level detected by flow cytometry.

Methodology:
It is a case control study enrolled 80 patients with multiple sclerosis at different stages and 70 cross matched healthy controls not suffering MS with similar age and demographic features.
Inclusion criteria:
• The diagnosis of MS was established clinically from the history according to revised McDonald’s criteria 2017.
Exclusion criteria:
• Patients proved not to have MS either at first interview or at any time during follow up were ruled out of the study.
• Healthy group with past medical history of autoimmune disease or other neurologic condition.
• Individuals within both groups were subjected to polymorphism typing of the selected SNP rs7582694 of STAT4 gene using PCR and detection of STAT4 proteins using flow cytometer.
Results:
• The age of the patients: ranged from 20-55 years (Mean+SD, 33.39 ± 8.00). According to sex sixty-nine was females (86.3%).
• The age of onset had median ranged between (18-49) years old regarding to subtype of MS and majority of patients (92.5%) had age of onset between eighteen and forty years old that was found in the same range between active and inactive groups.
• The main subtype was RRMS (82.5%), followed by SPMS (8.7%), CIS (5%) and PPMS (3.8%)
• EDSS score varies from 1 to 6.5 in the total patients with median of 3 with low level of clinical disability in RRMS patients (2.5), and CIS patients (1) and high level among SPMS patients (6) and among PPMS patients (6.5). That indicated progression of disability in SPMS and PPMS. Active disease was found among (45.0%) of the total patients, (45.5%) of the RRMS patients, (57.1%) of the SPMS patients and (50.0%) of the CIS patients.
• By brain MRI all RRMS and SPMS patients who had MRI examination were showing positive findings however in PPMS 1 case were negative and other 2cases were positive, on other side by spinal MRI all cases who had spinal MRI examination among SPMS and PPMS were positive and for RRMS 27 cases were positive and 22 were negative and four cases were CIS which diagnosed clinically and not by MRI.
• By using flow cytometer, the range of the STAT4 proteins detected by flow cytometry in the peripheral blood leukocytes; in the lymphocytes was (1.1 – 13.01), in the monocytes was (0.4 – 12.6) and in both lymphocytes and monocytes was (1.9 – 25.1) and regarding the controls, the STAT4 proteins were not found and couldn’t be detected by the flow cytometry (0.0). We found highly statistically significant increase in the level of STAT4 proteins in the active disease group compared to the inactive disease group (P=0.000): as higher values were reported in active group; these findings supported the role of STAT4 in pathogenesis of MS.
• In the same line we also used PCR to detect the distribution of alleles and genotypes of the selected SNPs of STAT4 gene (rs7582694) in all MS patients and controls.
• The distribution of genotypes and alleles rate in STAT4 detected by PCR showed significant increase in the percent of the GG genotype in controls compared to its percent in MS patients, while the GC and CC genotypes were more common in MS than controls. The frequency of the C allele was higher in MS patients than in control. The same relations found in its subtypes RRMS and SPMS. We found that patients with GC and CC variants were more likely to be associated with MS disease by univariate analysis and considered them as risk factors for MS disease. In the same line we found statistically significant increase in the percent of the GC and CC genotypes in the active group than inactive group. The frequency of the C allele was higher in the active disease group