Impact of EBV on Multiple in a Sample of Iraqi Females: Immunological and Molecular Study

Abstract

This study was designed to highlight the role of Epstein Barr viruses (EBV) as a possible causative effect in multiple sclerosis (MS) through testing the viral load along with both biochemical and immunological parameters in female MS patients. We also aimed at finding the effects of different types of treatment line on the various study aspects. The results of the enzyme-linked immunosorbent assay (ELISA) revealed first an increased level of EBV EBNA1IgG IL-17A (96.13±27.60 pg/ml) in sera of female MS patients (0.05±0.01 U/ml, n=50) compared with the control group (n=40). We confirmed this result using real-time polymerase chain reaction (RT-PCR) which also showed a significantly higher EBV load in MS patients (22.61±12.72 copies/ml) as compared to the control. This was associated with an increased level of IL-17A (96.13±27.60 pg/ml) in the patients, while levels of both melatonin hormone (0.63±0.19 pg/ml) and vitamin D3 (29.90±12.41 ng/ml) showed a significant decrease as compared to the normal values of the control group. Analysis of the relation between each of the studied parameters with the level of disability in the patients, as reflected by the expanded disability status score (EDSS), revealed no significant differences between the two categories of EDSS ˂3 and ≥3 patients in terms of EBV EBNA1IgG levels, whereas they were slightly different in terms of IL-17A and vitamin D3 (p <0.001). With respect to differences between the treatment lines, the results from the showed analysis of variance (ANOVA) test showed no significant differences in all the tested parameters between patients treated with the first line (Rebif, Avonex, Betaferon) and second-line (Gilenya, Tysabri). However, the only significant difference in comparison to the control group was found in levels of IL-17.
 It could be concluded that EBV has a crucial role in the initiation and targeting MS disease, leading to increasing levels of IL-17A, and decrease serum levels of melatonin hormone for female patients.

The Potential Role of Epstein-Barr Virus Nuclear Antigen-1 (Ebna-1) in Multiple Sclerosis

“…The significantly higher seropositivity of EBNA-1 IgG in the MS patients than in the controls is in accordance with other studies (20,21) , however, other studies, (Banwell et al, 2007), which was carried out on children, failed to manifest any relationship between the virus and MS (22) . The median level of EBNA-1 IgG for MS patients and controls were 81.08 U/ml and 67.73 U/ml respectively, which was significantly higher in MS patients in the current study, as reported by other studies which have showed significantly higher IgG level in MS patients (10,11,23,24) , while others found no significant difference (25,26) . Concerning the association between EBNA-1 IgG results and duration of MS disease, the current study revealed higher serum level and seropositivity of EBNA-1 IgG in MS patients during first two years of disease and this result is in accordance with other study (11) , this is indicative of the role of the viral antigen in the early stages of the disease which could be the triggering factor of MS. And due to this triggering factor, the relationship linking EBNA-1 IgG titers and number of clinical relapses showed no statistically significant association as shown in the table (5), this result is close to what's mentioned by Bakir et al in 2017 (10) .…”

“…On the other hand, in this study, patients with the first-line and second-line treatment showed no significant differences in EBNA 1 IgG levels between each other, while the median level in patients without treatment were higher, this result is comparable to result of study carried out by Abd Al Kareem and Abd in 2020 (11) , these patients who were recently diagnosed with MS had very high median IgG titer also supporting the possible triggering factor by the viral antigen.…”

“…In the current study, EBNA-1 IgG antibody was positive in 51.7% (62/121) of MS patients and 39.2% (47/121) of controls. To the best of our knowledge, there are two previous studies regarding the seroprevalence of EBNA-1 IgG in Iraq, the more recent study carried out by Abd Al Kareem and Abd in 2020 among the Iraqi female patients with MS who were admitted to Clinic of MS in Neuro-Science Hospital in Baghdad and they reported that mean serum level was significantly higher in female patients than in the healthy controls (11) , another study also show significantly higher level in MS patients than the healthy controls (Bakir et al, 2017), which were conducted in Rizgary Teaching Hospital in Erbil, Iraq (10) . The significantly higher seropositivity of EBNA-1 IgG in the MS patients than in the controls is in accordance with other studies (20,21) , however, other studies, (Banwell et al, 2007), which was carried out on children, failed to manifest any relationship between the virus and MS (22) .…”

“…Ascherio et al (9) found that IgG specific for EBV nuclear antigens were significantly elevated in plasma collected before the onset of MS and did not change significantly after MS onset. To the best of our knowledge, there are two previous studies regarding the seroprevalence of EBNA-1 IgG in Iraq, Bakir et al in 2017 showed significantly higher level in MS patients than the healthy controls and he was revealed the possible role of EBV during the course of MS autoimmune disease via antibody EBNA-1 IgG seropositivity and correlated with type of attack or relapse (10) , Abd Al Kareem and Abd in 2020 reported that mean serum level was significantly higher in female patients than in the healthy controls and they revealed that the pathogenesis of EBV has crucial role in initiation MS disease in females patients (11) . The objective of this study is to find out any correlation between MS development and the incidence and levels of anti-EBNA-1 IgG.…”

Impact of Interleukin-37 and Vitamin D on children with Streptococcus pyogenes sore throat in Salah al-Din Governorate, Iraq

“…Sore throat is a common presentation in pediatric office settings and emergency departments Bacterial infection due to Streptococcus pyogenes represented about 25% of sore throat in children ( Abd Al-Kareem et al, 2014). This a major human-specific bacterial pathogen that causes a wide array of manifestations ranging from mild localized infections to life-threatening invasive infections (Hadi et al,2017).…”