Dehee A, Asselot C, Piolot T, Jacomet C, Rozenbaum W, Vidaud M, Garbarg-Chenon A, Nicolas J C
Service de Virologie, E.A. 2391, Hôpital Trousseau, 26 rue du Dr. A. Netter, 75012 Paris, France.
J Med Virol. 2001 Nov;65(3):543-52.
Epstein-Barr virus (EBV) reactivation is more likely to occur in immunocompromised patients with subsequent higher susceptibility to EBV-associated lymphoproliferations. In contrast to transplant recipients, limited data are available concerning the EBV load in HIV-infected patients, with or without AIDS-related non-Hodgkin's lymphomas. We developed a TaqMan real-time PCR assay, allowing both the EBV genome and a cellular gene to be quantified in order to obtain a reliable normalized measurement of the EBV load in peripheral blood mononuclear cells (PBMCs). With a wide 6-log(10) quantification range and inter-assay variations of less than 24%, this quantitative PCR was sufficiently accurate and reproducible for routine follow-up. The EBV load was determined in PBMCs from 113 HIV-infected patients, 11 patients with primary HIV infection and 24 HIV-seronegative healthy controls. The rates of EBV detection were similar in the three groups. However, EBV loads were higher in the HIV-infected group (P < 0.00001) except for the patients with primary HIV infection. Unexpectedly, EBV loads were not correlated with the clinical stages of HIV infection or HIV replication, and did not depend on the degree of immunodepression, as judged by CD4+ counts. This study contributes towards the definition of the baseline EBV load during HIV infection and stresses the broad inter-individual variability of the EBV load in HIV-infected patients. Real-time PCR provides a useful tool that can be used in further longitudinal studies to assess the relevance of the EBV load to identify HIV-infected patients with a high risk of EBV-associated lymphoproliferations.
爱泼斯坦-巴尔病毒（EBV）再激活更有可能发生在免疫功能低下的患者中，这些患者随后对EBV相关的淋巴增殖性疾病易感性更高。与移植受者不同，关于感染HIV的患者（无论有无艾滋病相关非霍奇金淋巴瘤）的EBV载量，可用数据有限。我们开发了一种TaqMan实时荧光定量PCR检测方法，可同时对EBV基因组和一个细胞基因进行定量，以便获得外周血单个核细胞（PBMC）中EBV载量的可靠标准化测量值。该定量PCR的定量范围宽达6个对数（10），批间变异小于24%，对于常规随访而言足够准确且可重复。我们测定了113例感染HIV患者、11例原发性HIV感染患者和24例HIV血清阴性健康对照者的PBMC中的EBV载量。三组的EBV检测率相似。然而，除原发性HIV感染患者外，HIV感染组的EBV载量更高（P<0.00001）。出乎意料的是，EBV载量与HIV感染的临床分期或HIV复制无关，也不取决于免疫抑制程度（以CD4+细胞计数判断）。本研究有助于明确HIV感染期间的基线EBV载量，并强调了HIV感染患者中EBV载量存在广泛的个体间差异。实时荧光定量PCR提供了一种有用的工具，可用于进一步的纵向研究，以评估EBV载量对于识别有EBV相关淋巴增殖性疾病高风险的HIV感染患者的相关性。
