Quantitative Detection of Epstein-Barr Virus DNA in Cerebrospinal Fluid and Blood Samples of Patients with Relapsing-Remitting Multiple Sclerosis

Abstract

The presence of Epstein-Barr Virus (EBV) DNA in cerebrospinal fluid (CSF) and peripheral blood (PB) samples collected from 55 patients with clinical and radiologically-active relapsing-remitting MS (RRMS) and 51 subjects with other neurological diseases was determined using standardized commercially available kits for viral nucleic acid extraction and quantitative EBV DNA detection. Both cell-free and cell-associated CSF and PB fractions were analyzed, to distinguish latent from lytic EBV infection. EBV DNA was detected in 5.5% and 18.2% of cell-free and cell-associated CSF fractions of patients with RRMS as compared to 7.8% and 7.8% of controls; plasma and peripheral blood mononuclear cells (PBMC) positivity rates were 7.3% and 47.3% versus 5.8% and 31.4%, respectively. No significant difference in median EBV viral loads of positive samples was found between RRMS and control patients in all tested samples. Absence of statistically significant differences in EBV positivity rates between RRMS and control patients, despite the use of highly sensitive standardized methods, points to the lack of association between EBV and MS disease activity.

Epstein-Barr virus antibodies in serum and DNA load in saliva are not associated with radiological or clinical disease activity in patients with early multiple sclerosis

“…In keeping with previous findings [20, 22, 40, 41], EBV DNA was only very rarely detectable in whole blood of patients with CIS/early RRMS, again arguing against an impaired immunological control of EBV in patients with early MS.…”

“…While we did not detect an association of EBV DNA load in saliva and whole blood with MS disease activity, we cannot exclude that EBV reactivation in secondary lymphoid organs or within the brain could be associated with disease activity in MS. Nevertheless, patients with MS do not show increased frequencies or levels of EBV DNA in their cerebrospinal fluid [40, 41], rather arguing against the possibility that EBV reactivation in the CNS may predict MS disease activity. As we measured EBV IgG in serum and DNA in saliva only at a single point in time, our study does not permit to draw conclusions on the temporal dynamics of EBV parameters (i.e.…”

Epstein–Barr virus and multiple sclerosis: potential opportunities for immunotherapy

“…The insensitivity of DNA PCR for detecting EBV in PBMC or whole blood is demonstrated by the detection of EBV DNA in PBMC or whole blood in only 26.4–81.3% of MS patients 95 , 96 , 97 , 98 , 99 , 100 , 101 despite virtually all (>99%) MS patients being EBV seropositive 34 , 36 . The insensitivity of this approach is due to the fact that in healthy EBV‐seropositive subjects, the frequency of EBV‐infected cells within the peripheral blood B‐cell population is only ~5 per 10 6 B cells 107 .…”

“…EBV DNA has been detected in the CSF or cell pellets isolated from the CSF in only small proportions (0–12.5% and 18.2%, respectively) of MS patients, with no significant differences between MS patients and controls 99 , 100 , 102 , 104 , 115 . Even in patients with a demonstrated high level of EBV infection in B cells and plasma cells within brain tissue, the frequency of detection of EBV DNA in the CSF was low (12.5%) 115 .…”

“…114 EBV DNA has been detected in the CSF or cell pellets isolated from the CSF in only small proportions (0-12.5% and 18.2%, respectively) of MS patients, with no significant differences between MS patients and controls. 99,100,102,104,115 Even in patients with a demonstrated high level of EBV infection in B cells and plasma cells within brain tissue, the frequency of detection of EBV DNA in the CSF was low (12.5%). 115 The use of single-cell reverse transcription-PCR for EBER1 on individually sorted B cells and plasma cells from the CSF has yielded conflicting results, with one study detecting no EBV-infected cells 116 and another study finding an increased frequency of EBVinfected B cells in the CSF of patients with MS compared with patients with non-inflammatory neurological diseases.…”