From the Department of Immunology (L.D., J.D., N.C.), St James's Hospital; School of Medicine (M.S., N.C.), Trinity College Dublin; MS Unit (M.K., H.K.), Department of Neurology, St James's Hospital; Department of Medical Gerontology (M.M.), School of Medicine, Trinity Translational Medicine Institute, Trinity College Dublin; Virology Laboratory (Y.L., K.O.D., C.R., A.Q., B.C.), St James's Hospital; and FutureNeuro SFI Research Centre (H.K.), Academic Unit of Neurology, School of Medicine, Trinity College Dublin, Ireland.
Neurol Neuroimmunol Neuroinflamm. 2024 May;11(3):e200217. doi: 10.1212/NXI.0000000000200217. Epub 2024 Mar 28.
Epstein-Barr virus (EBV) has been strongly implicated in the pathogenesis of multiple sclerosis (MS). Despite this, there are no routinely used tests to measure cellular response to EBV. In this study, we analyzed the cellular response to EBV nuclear antigen-1 (EBNA-1) in people with MS (pwMS) using a whole blood assay.
This cross-sectional study took place in a dedicated MS clinic in a university hospital. We recruited healthy controls, people with epilepsy (PWE), and pwMS taking a range of disease-modifying treatments (DMTs) including natalizumab, anti-CD20 monoclonal antibodies (mAbs), dimethyl fumarate (DMF), and also treatment naïve. Whole blood samples were stimulated with commercially available PepTivator EBNA1 peptides and a control virus-cytomegalovirus (CMV) peptide. We recorded the cellular response to stimulation with both interferon gamma (IFN-γ) and interleukin-2 (IL-2). We also compared the cellular responses to EBNA1 with IgG responses to EBNA1, viral capsid antigen (VCA), and EBV viral load.
We recruited 86 pwMS, with relapsing remitting MS, in this group, and we observed a higher level of cellular response recorded with IFN-γ (0.79 IU/mL ± 1.36) vs healthy controls (0.29 IU/mL ± 0.90, = 0.0048) and PWE (0.17 IU/mL ± 0.33, = 0.0088). Treatment with either anti-CD20 mAbs (0.28 IU/mL ± 0.57) or DMF (0.07 IU/mL ± 0.15) resulted in a cellular response equivalent to control levels or in PWE ( = 0.26). The results of recording IL-2 response were concordant with IFN-γ: with suppression also seen with anti-CD20 mAbs and DMF. By contrast, we did not record any differential effect of DMTs on the levels of IgG to either EBNA-1 or VCA. Nor did we observe differences in cellular response to cytomegalovirus between groups.
This study demonstrates how testing and recording the cellular response to EBNA-1 in pwMS may be beneficial. EBNA-1 stimulation of whole blood samples produced higher levels of IFN-γ and IL-2 in pwMS compared with controls and PWE. In addition, we show a differential effect of currently available DMTs on this response. The functional assay deployed uses whole blood samples with minimal preprocessing suggesting that employment as a treatment response measure in clinical trials targeting EBV may be possible.
疱疹病毒 4 型（EBV）被强烈认为与多发性硬化症（MS）的发病机制有关。尽管如此，目前还没有常规用于测量细胞对 EBV 反应的检测方法。在这项研究中，我们使用全血检测分析了 MS 患者（pwMS）对 EBV 核抗原-1（EBNA-1）的细胞反应。
这是一项在大学医院专门的 MS 诊所进行的横断面研究。我们招募了健康对照组、癫痫患者（PWE）和接受多种疾病修正治疗（DMT）的 pwMS，包括那他珠单抗、抗 CD20 单克隆抗体（mAb）、二甲基富马酸（DMF），也包括治疗初治患者。全血样本用市售的 PepTivator EBNA1 肽和对照病毒-巨细胞病毒（CMV）肽刺激。我们记录了干扰素 γ（IFN-γ）和白细胞介素 2（IL-2）刺激的细胞反应。我们还比较了 EBNA1 的细胞反应与 EBNA1、病毒衣壳抗原（VCA）和 EBV 病毒载量的 IgG 反应。
我们招募了 86 名 pwMS，其中复发缓解型 MS 患者，我们观察到 IFN-γ记录的细胞反应水平更高（0.79 IU/mL ± 1.36），与健康对照组（0.29 IU/mL ± 0.90， = 0.0048）和 PWE（0.17 IU/mL ± 0.33， = 0.0088）相比。使用抗 CD20 mAb（0.28 IU/mL ± 0.57）或 DMF（0.07 IU/mL ± 0.15）治疗导致细胞反应与对照水平或 PWE 相当（ = 0.26）。记录 IL-2 反应的结果与 IFN-γ一致：抗 CD20 mAb 和 DMF 也观察到抑制作用。相比之下，我们没有记录到 DMT 对 EBNA-1 或 VCA 的 IgG 水平有任何差异作用。我们也没有观察到各组之间细胞对巨细胞病毒反应的差异。
这项研究表明，在 pwMS 中检测和记录对 EBNA-1 的细胞反应可能是有益的。与对照组和 PWE 相比，全血样本中 EBNA-1 的刺激产生了更高水平的 IFN-γ 和 IL-2。此外，我们还观察到目前可用的 DMT 对这种反应的差异影响。所使用的功能测定法使用全血样本进行，预处理最少，这表明它可能作为针对 EBV 的临床试验中的治疗反应测量方法。
