Long-term infection and shedding of human cytornegalovirus in T98G glioblastoma cells
Long-term infection and shedding of human cytornegalovirus in T98G glioblastoma cells
复制标题
DOI：
10.1128/jvi.00866-07
复制
发表时间：
2007-10-01
影响因子：
5.4
通讯作者：
Fortunato, Elizabeth A.
中科院分区：
文献类型：
--
作者：
Luo, Min Hua;Fortunato, Elizabeth A.
人巨细胞病毒（HCMV）是导致出生缺陷的主要病毒原因，主要影响中枢神经系统（CNS）。为了进一步理解这种CNS病理学，用HCMV感染来自胶质母细胞瘤细胞系T98 G和A172、星形胶质细胞胶质母细胞瘤细胞系CCF-STTG 1（CCF）和神经母细胞瘤细胞系SH-SY 5 Y（SY 5 Y）的细胞。CCF和SY 5 Y细胞对感染是完全允许的，而A172细胞是不允许的。在T98 G细胞中，大多数细胞在感染后6 h（hpi）显示病毒沉积到细胞核中;然而，在前72 h仅在30%的细胞中观察到病毒立即早期基因表达。在病毒抗原（Ag）阳性细胞中，虽然完整的病毒复制中心的发展被延迟，但完全发展的中心在96 hpi形成。有趣的是，即使在感染后很晚的时间，多个小，双极和大病灶的混合物总是存在。输入pp 65进入细胞核的初始运输也被延迟。滴度和感染中心测定显示少量T98 G细胞以非常低的水平脱落病毒。令人惊讶的是，Ag阳性和Ag阴性细胞都继续分裂;由于这种连续分裂，我们采用了每三天传代T98 G细胞的方案，以防止过度拥挤。在该方案下，可检测的感染性病毒脱落持续至第5代，病毒基因表达持续至第8代。这一证据表明T98 G细胞是一种有前途的长期感染模型。
Human cytomegalovirus (HCMV) is the leading viral cause of birth defects, affecting primarily the central nervous system (CNS). To further understand this CNS pathology, cells from glioblastoma cell lines T98G and A172, the astrocytic glioblastoma cell line CCF-STTG1 (CCF), and the neuroblastoma cell line SH-SY5Y (SY5Y) were infected with HCMV. CCF and SY5Y cells were fully permissive for infection, while A172 cells were nonpermissive. In T98G cells, the majority of cells showed viral deposition into the nucleus by 6 h postinfection (hpi); however, viral immediate-early gene expression was observed in only similar to 30% of cells in the first 72 h. In viral antigen (Ag)-positive cells, although the development of complete viral replication centers was delayed, fully developed centers formed by 96 hpi. Interestingly, even at very late times postinfection, a mixture of multiple small, bipolar, and large foci was always present. The initial trafficking of input pp65 into the nucleus was also delayed. Titer and infectious-center assays showed a small number of T98G cells shedding virus at very low levels. Surprisingly, both Ag-positive and Ag-negative cells continued to divide; because of this continuous division, we adopted a protocol for passaging the T98G cells every third day to prevent overcrowding. Under this protocol, detectable infectious-virus shedding continued until passage 5 and viral gene expression continued through eight passages. This evidence points to T98G cells as a promising model for long-term infections.