Investigation of pathologic factors of chronic active EB virus infection using microarray and human tissue model

为建立研究EB病毒（EBV）感染的模型系统，我们检测了B 95 -8病毒和表达增强型绿色荧光蛋白（EGFP-EBV）的重组EBV在人淋巴组织中的复制能力。将在常规扁桃体切除术中手术切除的人扁桃体组织切成小块，并置于空气界面处培养基中的胶原海绵凝胶顶部，然后将无细胞病毒悬液直接应用于每个组织块的顶部。在用B 95 -8和EGFP-EBV感染的组织模型中，在感染后12-15天至24天观察到培养基中EBV DNA水平增加。在培养过程中，从培养基中收集的细胞中的8个EBV相关基因的表达水平增加。此外，流式细胞术分析显示，大多数EGFP^+细胞为CD 3 ^-CD 56 ^-CD 19 ^+ HLA-DR^+，代表幼稚（IgD^+）和记忆（CD 27 ^+）B细胞。此外，在这个模型中，EBV复制抑制阿昔洛韦治疗剂量依赖性的方式。这些数据表明，该模型具有潜在的用于在原发感染时的局部组织的病理分析，以及用于筛选新的抗病毒剂。

To establish a model system for the study of Epstein-Barr virus (EBV) infection, we tested the ability of B95-8 virus and recombinant EBV expressing enhanced green fluorescent protein (EGFP-EBV) to replicate in human lymphoid tissue. Human tonsil tissues that had been surgically removed during routine tonsillectomy were sectioned into small blocks and placed on top of collagen sponge gels in culture medium at the air-interface, then a cell-free viral suspension was directly applied to the top of each tissue block. Increasing levels of EBV DNA in culture medium were observed after 12-15 days through 24 days post-infection in tissue models infected with B95-8 and EGFP-EBV. Expression levels of eight EBV-associated genes in cells collected from culture medium were increased during culture. Additionally, flow cytometry analyses revealed that most EGFP^+ cells were CD3^- CD56^- CD19^+ HLA-DR^+, and represented both nalve (IgD^+) and memory (CD27^+) B cells. Moreover, EBV replication in this model was suppressed by acyclovir treatment in a dose-dependent manner. These data suggest that this model has potential for use in the pathological analysis of local tissues at the time of primary infection, as well as for screening novel antiviral agents.