HCMV IE1和pp65在人神经胶质瘤U251细胞中的时序表达

目的:研究人神经胶质瘤U251细胞对人巨细病毒(human cytomegalovirus, HCMV)的容许性,并探讨病毒蛋白IE1及pp65的时序表达特点.方法:HCMV感染U251细胞,以透射电镜观察U251细胞的形态学改变,以PCR方法检测HCMV核酸,于不同的时间点分别用抗蛋白IE1及蛋白pp65单克隆抗体进行免疫细胞化学染色,检测IE1及pp65的表达水平.结果:受染的U251细胞出现了明显的形态学改变,并用PCR方法检测到了HCMV核酸.免疫细胞化学染色表明,蛋白IE1在感染后30min～2h无表达,4h开始表达,14h达高峰,随后下降;蛋白pp65在感染后1 h为入侵型pp65,4～96h一直呈低水平表达,至120h表达急剧升高.结论:U251细胞是HCMV的容许细胞,HCMV在其中的表达具有时序性,但是其时序表达较在人成纤维细胞中延迟.

Temporal expression of HCMV IE1 and pp65 in human glioma U251 cells

Objective To determine whether U251 cells are permissive for human cytomegalo-virus (HCMV), and to investigate the characteristics of temporal expression of proteins IE1 and pp65. Methods U251 cells were infected with HCMV, and then the cells were observed under the transmission electronic microscope, and the viral nucleic acid was detected by PCR, and the expression levels of IE1 and pp65 were analyzed by immunohistochemical assay with anti-IE1 monoclonal antibody and anti-pp65 monoclonal antibody at various time spost infection. Results Morphological changes of the infected cells appeared under the transmission electron microscope. The viral nucleic acid was detected successfully by PCR. The expression of IE1 was detected firstly at 4 h post infection, and reached a peak within 14 h, and then decreased. The incoming pp65 was detected at 1 h, the low expression levels of pp65 were detected firstly at 4 h, and they could remain relatively constant through 96 h, but the maximum expression occurred at 120 h. Conclusion Human glioma U251 cells are permissive for HCMV, the temporal cascade of HCMV gene expression can be observed in the infected U251 cells, but it is delayed obviously in the human fibroblast.