Department of Neurology, The University of Texas Southwestern Medical Center, Dallas2Department of Ophthalmology, The University of Texas Southwestern Medical Center, Dallas.
Laboratory of Molecular Medicine and Neuroscience, National Institutes of Health (NIH), Bethesda, Maryland.
JAMA Neurol. 2014 May;71(5):596-602. doi: 10.1001/jamaneurol.2014.63.
Infection with JC virus (JCV) may lead to development of demyelinating progressive multifocal leukoencephalopathy in patients with multiple sclerosis (MS) who are treated with natalizumab.
To determine whether mononuclear cells in circulation from MS patients treated with natalizumab harbor JCV DNA.
DESIGN, SETTING, AND PARTICIPANTS: In this prospective investigation, we enrolled 49 MS patients from the Clinical Center for Multiple Sclerosis at The University of Texas Southwestern Medical Center and 18 healthy volunteers. We drew 120-mL blood samples from 26 MS patients at baseline and at approximately 3-month intervals to 10 months during the course of natalizumab infusions. One blood sample was drawn from 23 MS patients receiving natalizumab for more than 24 months and from 18 healthy volunteers.
Natalizumab treatment of MS.
The blood samples were separated using flow cytometry into CD34+, CD19+, and CD3+ cell subsets; DNA templates were prepared using quantitative polymerase chain reaction for JCV DNA identification. Plasma samples were tested for anti-JCV antibodies by enzyme-linked immunosorbent assays performed at the Laboratory of Molecular Medicine and Neuroscience, National Institute of Neurological and Communicative Diseases and Stroke.
Thirteen of the 26 patients (50%) with baseline and follow-up blood samples had detectable viral DNA in at least 1 cell compartment at 1 or more points. Ten of the 23 patients (44%) receiving treatment for more than 24 months and 3 of the 18 healthy volunteers (17%) also had detectable viral DNA in 1 or more cell compartment. Fifteen of the 49 MS patients (31%) were confirmed to harbor JCV in CD34+ cells and 12 of 49 (24%) in CD19+ cells. Only 1 of 18 healthy volunteers were viremic in CD34+ cells and none in CD19+ cells. Nine patients and 1 healthy volunteer were viremic but had seronegative test results for JCV antibodies.
JC virus DNA was detectable within cell compartments of natalizumab-treated MS patients after treatment inception and longer. JC virus DNA may harbor in CD34+ cells in bone marrow that mobilize into the peripheral circulation at high concentrations. Latently infected cells initiate differentiation to CD19+ cells that favors growth of JCV. These data link the mechanism of natalizumab treatment with progressive multifocal leukoencephalopathy.
JC 病毒（JCV）感染可能导致接受那他珠单抗治疗的多发性硬化症（MS）患者发展为脱髓鞘进行性多灶性白质脑病。
确定接受那他珠单抗治疗的 MS 患者循环中的单核细胞是否携带 JCV DNA。
设计、地点和参与者：在这项前瞻性研究中，我们从德克萨斯大学西南医学中心的多发性硬化症临床中心招募了 49 名 MS 患者和 18 名健康志愿者。我们从 26 名 MS 患者中抽取了 120 毫升血液样本，这些患者在那他珠单抗输注过程中，在基线时和大约每 3 个月至 10 个月进行一次。从接受那他珠单抗治疗超过 24 个月的 23 名 MS 患者和 18 名健康志愿者中抽取了 1 份血液样本。
那他珠单抗治疗 MS。
使用流式细胞术将血液样本分离为 CD34+、CD19+和 CD3+细胞亚群；使用定量聚合酶链反应制备 DNA 模板以鉴定 JCV DNA。使用国立神经病学与中风研究所的分子医学和神经科学实验室的酶联免疫吸附试验检测血浆样本中的抗 JCV 抗体。
在基线和随访血液样本中，有 13 名患者（50%）至少在 1 个或多个时间点的 1 个或多个细胞隔室中检测到病毒 DNA。23 名接受治疗超过 24 个月的患者中有 10 名（44%）和 18 名健康志愿者中有 3 名（17%）在 1 个或多个细胞隔室中也检测到病毒 DNA。49 名 MS 患者中有 15 名（31%）在 CD34+细胞中被证实携带 JCV，49 名中有 12 名（24%）在 CD19+细胞中携带 JCV。18 名健康志愿者中只有 1 名在 CD34+细胞中病毒血症，而在 CD19+细胞中没有。9 名患者和 1 名健康志愿者病毒血症但 JCV 抗体检测结果阴性。
在那他珠单抗治疗开始后和更长时间内，可在 MS 患者的细胞隔室中检测到 JCV DNA。JC 病毒 DNA 可能存在于骨髓中的 CD34+细胞中，这些细胞在高浓度时会动员到外周循环中。潜伏感染的细胞分化为 CD19+细胞，有利于 JCV 的生长。这些数据将那他珠单抗治疗与进行性多灶性白质脑病的发病机制联系起来。
