Viral infections and multiple sclerosis

The Enigmatic Role of Viruses in Multiple Sclerosis: Molecular Mimicry or Disturbed Immune Surveillance?

We examined cerebral spinal fluid (CSF) from multiple sclerosis (MS) patients and patients with other neurological diseases (OND) for antibody specific for Human Herpesvirus-6 (HHV-6) and for HHV-6 DNA detectable by PCR. CSF from MS patients had a higher frequency of IgG antibody to HHV-6 late antigens (39.4%) compared with CSF from OND (7.4%). In contrast, the frequency of detectable IgG antibody in CSF from MS patients specific for Epstein-Barr Virus (EBV) (12.1%) and Human Cytomegalovirus (HCMV) (6.1%) was much lower. Two of 12 MS CSFs (16.7%) also contained HHV-6 DNA detected by PCR. None of four OND CSF were positive for HHV-6 DNA. Plasma from 16 patients with MS, eight with OND and 72 healthy donors were tested for antibodies by ELISA to HHV-6 early (p41/38) and late (gp110) proteins. Although no differences in anti-gp110 IgG antibody were detected between MS patients, patients with other neurological diseases, and normals, IgG antibody to early protein p41/38 was detected in > 68% of the plasma from MS patients, 12.5% from OND patients and 27.8% of the controls. IgM antibody to p41/38 was present in > 56% of MS patients, 12.5% of OND patients, and 19% of controls. These data suggest that more than half of the MS patients had active, ongoing HHV-6 infections. HHV-6 was also isolated from peripheral blood mononuclear cells (PBMC) from 3/5 MS patients who were in relapse or had progressive disease and was identified as HHV-6 Variant B. These preliminary results support the hypothesis that HHV-6 may be a co-factor in the pathogenesis of some cases of MS.

Objective Epstein-Barr virus (EBV) and Human Herpes virus 6 (HHV-6) are believed to involve in multiple sclerosis (MS) pathogenesis. Natural killer (NK) and CD8+ T cells have essential roles in handling viral infections and their phenotypic and functional properties may be influenced following exposure to viral infections. Here, we investigated the association of NK and CD8+ T cells subpopulations frequency with EBV and HHV-6 viral load in MS patients. Materials and Methods In this case-control study, EBV and HHV-6 viral load were evaluated in plasma of newly diagnosed relapsing-remitting MS (RRMS) patients at relapse phase (n=23), who were not on disease-modifying therapy (DMT), and sex- and age-matched healthy controls (n=19) using real-time polymerase chain reaction (PCR). The frequency of NK and CD8+ T cells subsets were assessed by CD27, CD28, CD45RO, CD56, and CD57 markers using flow cytometry. Results Despite the increased level of EBV viral load in RRMS patients compared to the control group, there was no statistically significant difference in EBV and HHV-6 copy numbers between the studied groups. In addition, a significant decrease was observed in the percentages of CD56brightCD57- and CD56dimCD57+CD8lowCD45RO- NK cells in RRMS patients in comparison to healthy controls. Analysis of CD8+ T cell subsets showed a substantially high proportion of CD27+CD28+CD45RO+CD57-CD8hiT cells in patients at relapse phase compared to controls. The frequency of NK and T cells subtypes was not associated with EBV and HHV6 plasma viral loads. ConclusionThese findings further highlight the variation of NK and CD8+ T cells subsets frequency in clinically active RRMS patients. Since the composition of cells was not associated with EBV and HHV-6 viral load, perhaps other viral infections may be involved in altered NK and CD8+ T cells subpopulation. Larger cohort studies are needed to confirm these results.

Introduction: Multiple sclerosis (MS) is an autoimmune neuroinflammatory and neurodegenerative disease that infects and destroys the central nervous system (CNS). Many variables influence the start of multiple sclerosis disease. MS was thought to be mostly caused by viral infection, particularly infections with the human herpes virus 6 (HHV-6), Epstein-Barr virus (EBV), and other viruses. Aim: the goal of the present study is to estimate the role of human herpesvirus-6 infection as a trigger factor for multiple sclerosis disorder and the role of some proinflammatory cytokine in early detection of this disorder. Material and Methods: We measured the titer of IgM, IgG Ab for human herpesvirus-6 and proinflammatory tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), between October 2023 and February 2024, in the 90 blood samples that were drawn from individuals aged 13 to 75. The medical personnel of Dr. Saad Al-Witry Hospital for Neurosciences provided diagnoses for these patients. We divided the subjects into 3 groups: the first group included 27 patients who suffered from multiple sclerosis with HHV-6 infection, the second group included 33 patients who suffered from multiple sclerosis without HHV-6, and the third group included 30 people who appeared to be in good health. We used the Enzyme-Linked Immunosorbent Assay (ELISA) technology to perform measurements. Results: The statistical analysis showed a significant increase (P≤0.01) in anti-HHV-6 IgM and IgG antibodies in the sera of patients with MS diseases and HHV-6 compared to the control group. There was also a significant increase (P≤0.01) in MS subjects without HHV-6 compared to the control group. Additionally, the statistical analysis of TNF-α level revealed a highly significant difference between MS patients with HHV-6 and those without HHV-6 compared with the control group. Furthermore, the statistical analysis showed a significant elevation (P≤0.01) in interleukin-6 (IL-6) in sera of patients with MS and HHV-6 compared to the control group and in MS subjects without HHV-6 compared to the control group. Conclusion: According to the current research, HHV-6 infection may be a major factor in MS

To assess the presence of Epstein-Barr virus (EBV) and human herpesvirus 6B (HHV-6B) DNA in saliva and plasma from multiple sclerosis (MS) patients enrolled in a randomized, double-blind, placebo-controlled valacyclovir treatment study. DNA was prepared following ultracentrifugation of saliva and plasma. EBV and HHV-6B DNAs were determined by real-time polymerase chain reaction. EBV and HHV-6B DNAs were detected in 41% and 65% of saliva samples, respectively. In patients treated with valacyclovir, the percentage of saliva samples with EBV was significantly reduced (9%; P = 0.000017), whereas the frequency of HHV-6B positive samples was unchanged (57%; P = 0.38). Longitudinal studies demonstrated a time-dependent reduction in the frequency of saliva samples containing EBV following valacyclovir treatment. In contrast, plasma contained EBV and HHV-6B DNAs in 17% and 25% of the samples, respectively, and these numbers were not significantly reduced following valacylovir treatment (13% and 16%, respectively), nor were they different from those of healthy controls (6% and 39%, respectively). Patients with high disease activity had a significantly higher frequency of EBV (P = 0.018) and HHV-6B (P = 0.023) positive samples than did patients with low disease activity. The presence of EBV and HHV-6B was strongly correlated in plasma (P < 0.00000001), but not in saliva (P = 0.41). MS patients express EBV and HHV-6B in both saliva and plasma, but only the expression of EBV in saliva is significantly reduced following valacyclovir treatment. Although EBV and HHV-6B DNAs can be detected in plasma from healthy individuals, the co-expression of both these viruses in MS patients is highly significant and further associated with clinical activity. The observations of viral DNA in plasma are consistent with an underlying immunologic defect in MS.

Higher frequency of Human herpesvirus-6 (HHV-6) viral DNA simultaneously with low frequency of Epstein-Barr virus (EBV) viral DNA in a cohort of multiple sclerosis patients from Rio de Janeiro, Brazil.

Background and Objective. Multiple sclerosis (MS) is a progressive disease of the nervous system that leads to demyelination of the nerves and is more common in young adults, especially women. Environmental factors that trigger MS pathogeneses are genetic susceptibility and viral infections. The viral infections are considered to be of particular relevance. Along with viruses belonging to the Herpesviridae family such as varicella zoster virus (VZV), human herpes virus-6 (HHV-6), and particularly Epstein–Barr virus (EBV), the expression of the env gene of human endogenous retrovirus (HERV-W) like MSRV is expected to be one of the risk factors for bringing up MS and disease progression. The present study aimed to investigate the correlation between EBV infection and HERV-W env in a case group (MS patients) and a control group (healthy individuals). Materials and Methods. 130 subjects were enrolled in a case-control study at two tertiary university hospitals from Tabriz (Imam and Razi), Iran. Of these, 65 subjects were MS patients serving as the case group, and 65 subjects were healthy individuals serving as the control group. After DNA extraction from all samples, the EBER region of EBV genome was used as the primer for the detection of EBV. RNA was extracted from PBMCs, and cDNA synthesis was performed by using Sina Gene kit. Subsequently, each sample was analysed by RT-PCR with two sets of primers to detect specifically multiple sclerosis retroviruses (MSRV) env, and RT-PCR was repeated for each HERV-W env. Positive sample was used in order to confirm the result. Results. In the case group, 19 (29.2%) patients were male and 46 (70.8%) patients were female. Nevertheless, in the control group, 21 (32.3%) subjects were male and 44 (67.7%) subjects were female. No significant difference was found between groups in gender (p = 0.70). The mean range in control and case groups was 33/38 ± 9/85 and 33.18 ± 8.65, respectively. No significant difference was found between groups in age (p = 0.902). 4 (6.2%) patients in case groups were found to be positive for EBV DNA (p = 0.119). Expression of the env gene of HERVs was observed in 10 (15.38%) and two (3.07%) specimens in the case and control groups (p = 0.030), separately. A comparison of the prevalence of the HERV ENV genome between the two study groups showed a significant difference (p = 0.005). Conclusion. The results of this study failed to show any difference between MS patients and healthy controls in the rate of EBV infection. It can be concluded that the expression of HERV-W/env genes may be involved in the development of MS in these patients.

Association between human herpesviruses and multiple sclerosis: A systematic review and meta-analysis

EBV Reactivation Incidence and Severity in Multiple Sclerosis Patients Treated with Autologous Haematopoietic Stem Cell Transplant and Comparing with Aplastic Anaemia Treated with ATG—Single Site Experience

Human herpesvirus 6 (HHV-6) and Epstein-Barr virus (EBV) have been repeatedly associated with multiple sclerosis (MS) pathogenesis. Also, it has been speculated that, besides its immunomodulatory properties, the efficacy of interferon beta (IFN-beta) in treating the disease may be related to its antiviral properties. The objectives of this study were to evaluate the in vivo antiviral effect of IFN-beta-1b over HHV-6 and EBV and to analyze whether such effect could be involved in its effectiveness in MS. A total of 54 patients with MS were included in an observational, multicentric, 24-month study. HHV-6 and EBV were detected by qPCR in blood and serum samples. IFN-beta-1b effectiveness was evaluated by presence, number and severity of relapses, reduction in the relapse rate, disability progression, and response to the treatment. Patients with HHV-6 in blood had a higher risk of severe relapses (P=0.01) and bad response (P=0.03). HHV-6 was detected more frequently during relapses than in remission in blood (P=0.024) and in serum (P=0.0002). Patients with HHV-6 in serum had more relapses (P=0.02), lesser reduction in the relapse rate (P=0.04), and a lower proportion of responders (P=0.02) than those without HHV-6 active replication. However, any association between EBV and clinical parameters could not be found. We concluded that presence of HHV-6 in blood and serum during IFN-beta treatment could be a good marker of poor response.

Multiple sclerosis (MS)-associated retrovirus (MSRV)/HERV-W (human endogenous retrovirus W) and Human herpesvirus 6 (HHV-6) are the two most studied (and discussed) viruses as environmental co-factors that trigger MS immunopathological phenomena. Autopsied brain tissues from MS patients and controls and peripheral blood mononuclear cells (PBMCs) were analysed. Quantitative RT-PCR and PCR with primers specific for MSRV/HERV-W env and pol and HHV-6 U94/rep and DNA-pol were used to determine virus copy numbers. Brain sections were immunostained with HERV-W env-specific monoclonal antibody to detect the viral protein. All brains expressed MSRV/HERV-W env and pol genes. Phylogenetic analysis indicated that cerebral MSRV/HERV-W-related env sequences, plasmatic MSRV, HERV-W and ERVWE1 (syncytin) are related closely. Accumulation of MSRV/HERV-W-specific RNAs was significantly greater in MS brains than in controls (P=0.014 vs healthy controls; P=0.006 vs pathological controls). By immunohistochemistry, no HERV-W env protein was detected in control brains, whereas it was upregulated within MS plaques and correlated with the extent of active demyelination and inflammation. No HHV-6-specific RNAs were detected in brains of MS patients; one healthy control had latent HHV-6 and one pathological control had replicating HHV-6. At the PBMC level, all MS patients expressed MSRV/HERV-W env at higher copy numbers than did controls (P=0.00003). Similar HHV-6 presence was found in MS patients and healthy individuals; only one MS patient had replicating HHV-6. This report, the first to study both MSRV/HERV-W and HHV-6, indicates that MSRV/HERV-W is expressed actively in human brain and activated strongly in MS patients, whilst there are no significant differences between these MS patients and controls for HHV-6 presence/replication at the brain or PBMC level.

Background: Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system that leads to inflammation, demyelination and neurodegeneration. Viral aetiology has been suspected to be an MS trigger for a long time, and herpesviruses (HSs) are among the potential pathogens involved. Objectives: The present investigation aims to detect the presence of antibodies against the herpes simplex virus (HSV), varicella-zoster virus, Epstein-Barr virus (EBV), human cytomegalovirus (CMV) and human herpesvirus 6 (HHV6) in the serum of MS patients and control individuals in north-eastern Poland. Method: Plasma was collected from 141 MS patients and 44 blood donors who served as the control group. These individuals were assessed for the presence of antibodies using an enzyme-linked immunosorbent assay. Results: The statistical analysis showed a higher probability of EBV (p = 0.037, OR 4.359) and HHV6 (p = 0.020, OR 3.343) antibody presence in patients with MS compared to that in the control group. In the MS patient group, the prevalence of CMV IgG antibodies was significantly higher in females (p = 0.025). Patients who tested positive for anti-EBV IgG were diagnosed 7.9 years earlier than patients who tested negative for anti-EBV IgG (p = 0.048). Conclusions: The study showed that MS patients in north-eastern Poland were more likely to be seropositive for EBV and HHV6 than healthy individuals. Further work should be undertaken in other regions of Poland and other European countries with particular attention paid to testing seropositivity in all HSs, particularly in the MS patient population, to evaluate the impact of HSs on MS patients in different environments.

Background Multiple sclerosis (MS) is an immune mediated inflammatory disease that attacks myelinated axons in the central nervous system (CNS), destroying the myelin and the axon in variable degrees. The aetiology and pathogenesis of MS is complex and multifactorial, involving many interlacing mechanisms. Many theories had considered viral infections as a possible cause of MS. One of these viruses is Epstein-Barr virus (EBV), a herpes virus belonging to the family herpesviridae. There is obvious similarity between EBV and MS regarding their epidemiological pictures, and it was observed that most MS patients had a history of infectious mononucleosis (IM) a few years before onset. The relation between EBV and MS may give hope for development of biomarkers for prediction of disease development, early diagnosis, prediction of prognosis, curing or even preventing MS through an anti EBV vaccine or antiviral therapies. Objectives This study aims to analyze the association between EBV infection and multiple sclerosis. Subjects and Methods This is a case control study carried in the MS outpatient clinic at Ain Shams University Hospitals during the period from April 2019 till November 2019. Subjects included in this study were classified into two groups. The first group included 30 patients diagnosed as having MS, on the basis of their MRI finding, clinical presentation and according to revised Mc Donald criteria 2017. The patient group included 11 males and 19 females, their age ranged from 18 to 48 years. The second group included 30 age and sex matched healthy controls without any neurological or medical diseases. The control group included 11 males and 19 females, their age ranged from 18 to 48 years. Both groups were tested quantitatively for immunoglobulin G against Epstein Barr viral capsid antigen (VCA) using the enzyme linked immunosorbent assay technique (ELISA). Results All patients with MS (100%) were positive for EBV VCA IgG, whereas (93.33%) of controls were positive. In the MS group, the EBV VCA IgG mean level was (161± 66.32) U/ml compared with (78.53±47.63) U/ml in controls. The difference in serum level of EBV VCA IgG between both groups was statistically highly significant (P = &amp;lt; 0.001). Conclusion There were higher levels of EBV VCA IgG in the serum of MS patients compared to healthy controls. This finding postulates a relation between EBV infection and MS and its role in the pathogenesis of MS.

Relevance.There is no predominant agent in the etiological structure of hemorrhagic vasculitis; instead, several factors play a role, including the activation of herpesvirus infections in the disease development. The objective of the study is determined by contradictory data and a lack of coordinated agreement regarding the etiology of hemorrhagic vasculitis. Objective: To identify the role of herpesvirus infection in the etiology of hemorrhagic vasculitis in children. Materials and methods. A cross-sectional study was conducted for 25 children aged from 2 months to 18 years with hemorrhagic vasculitis, who were on inpatient treatment at Medical Center "Children's Hospital" in Aktobe. All children underwent an enzyme-linked immunosorbent assay (ELISA) for herpesviruses: herpes simplex virus types I, II, (HSV-I, II), Epstein - Barr virus (EBV), cytomegalovirus (CMV), human herpesvirus type 6 (HHV – 6). The received data is processed by descriptive statistics, STATISTICS 10.0. Results The herpesvirus infection was confirmed in 100% of the examined children, with herpes simplex virus I and II types being discovered in 25.9% of cases, Epstein-Barr virus in 28.46% of cases, cytomegalovirus (CMV) in 29.6% of cases, and human herpesvirus 6 (HHV-6) in 16% of cases. Moreover, they did not occur as mono-infections but rather in conjunction with the cytomegalovirus: CMV+ HSV-I, II (16 %); CMV+ EBV (8%); CMV+ HHV-6 (4%); CMV+ EBV+ HHV-6 (32%); CMV+ HSV-I, II + EBV+ HHV-6 (40%). A low concentration of herpesvirus antibodies in the blood of patients with hemorrhagic vasculitis was detected by ELISA anti-CMV IgG (11.62 U / ml), anti-HHV-6 IgG (6.82 U/ml), which indicates unstable immunity, the risk of activation of viral infection and recurrent hemorrhagic vasculitis. Conclusion.According to the study, herpesvirus infections can lead to hemorrhagic vasculitis in children. If vasculitis recurs, it is advised to check for the presence of antibodies to herpesvirus antigens; if so, an infectious neurologist's consultation and etiotropic antiviral therapy are advised. Актуальность. В этиологической структуре геморрагического васкулита нет преобладания какого-либо одного агента, а играет роль совокупность нескольких факторов, в том числе активация герпесвирусных инфекций в развитии болезни. Противоречивые данные, отсутствие единого мнения об этиологии геморрагического васкулита определяет цель исследования. Цель. Определить роль герпесвирусной инфекции в этиологии геморрагического васкулита у детей. Материалы и методы. Одномоментное поперечное исследование проведено 25 детям с диагнозом геморрагический васкулит в возрасте от 2 мес. до 18 лет, которые находились на стационарном лечении в Актюбинском Медицинском Центре «Детском стационаре» г. Актобе. Всем детям проводился иммуноферментный анализ (ИФА) на герпесвирусы: вирус простого герпеса I, II-го типов, (HSV- I, II), вирус Эпштейна – Барр (EBV), цитомегаловирус (CMV), вирус герпеса человека 6-го типа (HHV-6). Полученные данные обработаны описательной статистикой, СТАТИСТИКА 10.0. Результаты. Герпесвирусная инфекция у обследуемых детей подтверждена в 100% случаев, из них вирус простого герпеса І, ІІ-го типов выявлен в 25,9%, вирус Эпштейна – Барр – 28,46%, цитомегаловирус (CMV) – 29,6%, вирус герпеса человека 6-го типа (HHV-6) – 16% случаев. И они не встречались как моноинфекции, а в виде ассоциации с цитомегаловирусом: CMV+ HSV-I, II (16 %); CMV+ EBV (8%); CMV+ HHV-6 (4%); CMV+ EBV+ HHV-6 (32%); CMV+ HSV-I, II + EBV+ HHV-6 (40 %). Выявлено малая концентрация антител герпесвирусов в крови больных с геморрагическим васкулитом методом ИФА анти-CMV IgG (11,62 ЕД/мл), анти-HHV-6 IgG (6,82 ЕД/мл), что свидетельствует о неустойчивом иммунитете, риске активации вирусной инфекции и рецидивирующим течением геморрагического васкулита. Заключение. Исследование показало роль герпесвирусной инфекций в развитии геморрагического васкулита у детей и при рецидивирующем васкулите рекомендуется обследование на наличие антител на антигены герпесвирусов и при их положительном результате рекомендуется консультация инфекциониста о решении этиотропной противовирусной терапии. Өзектілігі. Геморрагиялық васкулиттің этиологиялық құрылымында бір агент басым болмайды, бірақ бірнеше факторлардың жиынтығы, соның ішінде аурудың дамуындағы герпесвирустық инфекциялардың белсендірілуі маңызды рөл атарады. Қарама-қайшы дәлелдер, геморрагиялық васкулиттің этиологиясы туралы бірдей тұжырымның болмауы зерттеу мақсатын анықтайды. Мақсаты. Балалардағы геморрагиялық васкулиттің этиологиясындағы герпесвирустық инфекцияның рөлін анықтау. Материалдар мен әдістер. Ақтөбе қаласының Ақтөбе Медициналық орталығы Балалар стационарында" геморрагиялық васкулитпен стационарлық ем қабылдаған 2айдан 18 жасқа дейінгі 25 балаға бір мезгілде көлденең зерттеу жүргізілді. Барлық балаларға герпесвирустарға иммуноферменттік талдау (ИФА) жүргізілді: I, II типті қарапайым герпес вирусы, (HSV-I, II), Эпштейн - Барр вирусы (EBV), цитомегаловирус (CMV), 6 типті адамның герпес вирусы (HHV–6). Алынған мәліметтер сипаттамалық статистикамен өңделді, СТАТИСТИКА 10.0. Нәтижелері. Зерттелеген балалардағы герпесвирустық инфекция 100% жағдайда расталды, оның ішінде І, ІІ типті қарапайым герпес вирусы 25,9%, Эпштейн – Барр вирусы – 28,46%, цитомегаловирус (CMV) – 29,6%, 6 типті (HHV-6) адамның герпес вирусы -16% жағдайлар анықталды. Олар моноинфекция ретінде емес, цитомегаловируспен байланыс ретінде пайда болды: CMV+ HSV-I, II (16 %); CMV+ EBV (8%); CMV+ HHV-6 (4%); CMV+ EBV+ HHV-6 (32%); CMV+ HSV-I, II + EBV+ HHV-6 (40 %).Геморрагиялық васкулитпен ауыратын науқастардың қанында герпесвирус антиденелерінің төмен концентрациясы анти-CMV IgG (11,62 бірл/мл), анти-HHV-6 IgG (6,82 бірл/мл) ИФА әдісімен анықталды, бұл тұрақсыз иммунитетті, вирустық инфекцияны белсендіру қаупін және геморрагиялық васкулиттің қайталанатын ағымын көрсетеді. Қорытынды. Зерттеу балалардағы геморрагиялық васкулиттің дамуындағы герпесвирустық инфекциялардың рөлін көрсетті және қайталанатын васкулит кезінде герпесвирус антигендеріне антиденелердің болуын тексеру ұсынылады және олардың оң нәтижесі болған кезде этиотропты вирусқа қарсы терапияны шешу үшін инфекционистпен кеңес жүргізу ұсынылады.

Ultrasensitive detection and quantification of viral nucleic acids with Raindance droplet digital PCR (ddPCR).