【摘要】 目的研究HCMV感染HEL细胞后,病毒蛋白IE1及pp65在不同时间点的亚细胞分布及其表达量-时效变化相关性。方法HCMV感染HEL细胞后,吸附2h,维持培养0、2、4、6、10、22及46h后,用病毒蛋白IE1的单克隆抗体及蛋白pp65的单克隆抗体做免疫细胞化学染色,检测此两种蛋白的胞内分布;同时运用图像分析系统对此检测结果进行灰度值测定,并利用方差分析分析此测定结果。结果IE1蛋白仅在核内检测到;表达趋势是4~6hp.i.呈表达上升趋势,6~8hp.i.又下降。pp65首先于仅在核内被检测到,之后也分布于胞浆中。pp65表达于2~8hp.i.表达增多,8~24hp.i.呈胞内稳定表达,24~48hp.i.表达降低。结论IE1蛋白及pp65表达的亚细胞分布具有时空特异性;两种蛋白的表达量上都表现为量-时效相关性;本研究间接反映了病毒复制增殖的连续过程。更多还原

【Abstract】 Objective To study the sub-cellular distribution and the expression trends of HCMV proteins: the temporal-spacial relationship between IE1 and pp65 in the infected HEL cells. Methods Seven time-points were selected as follows: HEL cells were adsorbed by HCMV for 2 hours and then cultivated with the maintenance media for 0, 2, 4, 6, 10, 22 and 46h. By using monoclonal IE1 antibody and monoclonal pp65 antibody, the two proteins’ sub-cellular distribution and their expressions were examined immunohistochemically with an image analysis system. Results The IE1 protein was expressed in the nuclei only. The trend of its expression was ascended from 4 to 6h p.i. and then decreased quickly in 8h p.i.; pp65 was also first found in the nuclei and then also in the cytoplasm, which ascended to the peak within 6h p.i., maintained until 24h p.i. and descended from 24h p.i. to 48h p.i.. Conclusions The sub-cellular distributions of viral protein IE1 and pp65 showed a temporal-spacial relationship. The amount of both proteins varied with time. The experiment indirectly proves that the replication of HCMV in HEL cells is continuous.更多还原