EBV and vitamin D status in relapsing-remitting multiple sclerosis patients with a unique cytokine signature

Abstract

Multiple sclerosis, a debilitating autoimmune and inflammatory disease of the central nervous system, is associated with both infectious and non-infectious factors. We investigated the role of EBV infection, vitamin D level, and cytokine signature in MS patients. Molecular and serological assays were used to investigate immune biomarkers, vitamin D level, and EBV status in 83 patients with relapsing-remitting multiple sclerosis and 62 healthy controls. In total, 98.8 % of MS patients showed a history of EBV exposure compared to 88.6 % in the healthy group (p = 0.005). EBV DNA load was significantly higher in MS patients than healthy subjects (p < 0.0001). Using a panel of biomarkers, we found a distinct transcriptional signature in MS patients compared to the healthy group with mRNA levels of CD73, IL-6, IL-23, IFN-γ, TNF-α, IL-15, IL-28, and IL-17 significantly elevated in MS patients (p < 0.0001). In contrast, the mRNA levels for TGF-β, IDO, S1PR1, IL-10, and CCL-3 were significantly lower in MS patients compared to healthy controls (p < 0.0001). No significant differences were found with the mRNA levels of IL-13, CCL-5, and FOXP3. Interestingly, in MS patients we found an inverse correlation between vitamin D concentration and EBV load, but not EBNA-1 IgG antibody levels. Our data highlight biomarker correlates in MS patients together with a complex interplay between EBV replication and vitamin D levels.

Multiple sclerosis‐associated retrovirus, Epstein‐Barr virus, and vitamin D status in patients with relapsing remitting multiple sclerosis

“…25‐Hydroxy vitamin D levels were measured in sera by the IDS EIA kit (IDS Ltd, UK) according to the manufacturer's instructions. EBNA‐1 specific IgG antibodies were also determined using ELISA (Abnova, Monterotondo, Italy) as previously described …”

“…Although yet to be confirmed, infection with certain herpes viruses particularly Epstein‐bar virus (EBV) and human herpesvirus 6 (HHV‐6) were found to induce transcriptional activation of HERV, supporting the view that co‐infection with another virus may facilitate the development of Multiple Sclerosis . The importance of the concomitant effects of EBV infection and vitamin D deficiency, as important environmental risk factors, has been highlighted by previous studies . Interestingly, an inverse correlation has been recently reported between EBV DNA load and serum concentration of vitamin D in Multiple Sclerosis patients …”

“…The importance of the concomitant effects of EBV infection and vitamin D deficiency, as important environmental risk factors, has been highlighted by previous studies . Interestingly, an inverse correlation has been recently reported between EBV DNA load and serum concentration of vitamin D in Multiple Sclerosis patients …”

The interplay between vitamin D and viral infections

“…Expression of VDRs in almost all immune cells indicates that these are one of the main target of vitamin D, and numerous immune biomarkers are modulated via VDRs action (box 2). (10) given the fact that immune cells also express CYP27B1, it can locally facilitate the conversion of 25(OH) D to 1,25(OH)2D. Green color: Induction or increase of expression by vitamin D. red color: Inhibition or decreases of expression by vitamin D. abbreviation: 7-DHC, 7-dihydrocholesterol (7-DHC); 1,25 di-hydroxy-vitamin D [1,25(OH)2D], 25 hydroxy-vitamin D [25(OH)D], cytochrome P450 family 27 subfamily B member 1 (CYP27B1), cytochrome P450 family 24 subfamily a member 1 (CYP24A1), cytochrome P450 family 2 subfamily R member 1 (CYP2R1), retinoic X receptor (RXR), toll-like receptor (TLR), vitamin D receptor (VDR), vitamin D responsive elements (VDRE) and T cells to microbial invasion sites ( Figure 2).…”

Viruses and endogenous retroviruses in multiple sclerosis: From correlation to causation

“…35 With regard to vitamin D, some studies have failed to detect a statistically significant interaction, 32,36 while others have reported an interaction with either EBV antibodies or DNA load. 37 Mechanistically, observations that there is overlap between EBNA-2a and vitamin D receptor (VDR) binding sites within MS-associated genomic regions and that EBNA-3 binds to the VDR may provide further insights. 24,38 In neuroimaging studies, MRI (magnetic resonance imaging) markers of MS activity and grey matter atrophy were found to be associated with anti-EBV antibody levels.…”