摘要
目的明确人巨细胞病毒(HCMV)磷酸基质蛋白65(pp65)抗原在阳性细胞中的定位并初探其定位与病毒活动间的关系,为指导临床病毒抗原血症的监测提供依据。方法利用4,6-联脒-2-苯基吲哚(DAPI)荧光染料能与细胞双链DNA结合的特点,在激光共聚焦显微镜下对HCMV pp65单抗阳染细胞进行抗原定位;同时通过外周血白细胞与病毒共培养技术,检测分析其定位与病毒活动的关系。结果临床标本检测的HCM- Vpp65抗原阳染细胞的表现形式有3种,即细胞浆型、细胞核型和细胞浆核混合型,其中细胞核中的HCMV pp65抗原聚集于细胞核的内部,而非仅仅附着于核表面。进一步的白细胞与病毒共培养结果显示,只有野生型病毒株感染的人胚肺成纤维细胞(HELF)与外周血白细胞共培养,才能检出HCMV pp65抗原阳性的白细胞,而且表现为仅初期(培养1 h内)可检出细胞浆阳染型,随着时间延长,约80%～90%的抗原阳染白细胞表现为细胞核型,偶见细胞浆核混合型。结论HCMV pp65抗原血症时阳染的外周血白细胞的表现形式有3种,即细胞核型、细胞浆型以及细胞浆核混合型,其中细胞核型属细胞核内部型,并非细胞核表面附着型。结合共培养结果认为,细胞核型阳染细胞的检出更能提示病毒的活动。
Objective To identify the localization of human Cytomegalovirus（HCMV） pp65 antigen in the positive leukocytes and discuss the relation between the localization and activity of HCMV infection, to help the guide of HCMV pp65 antigenemia test. Methods By the laser scanning confocal microscopy, to detect the pp65 antigen localization in the positive peripheral leukocytes, and to analyze the relation between the localization of pp65 antigen and the infectivity of HCMV by cocuhure with HCMV infected human embryonic lung fibroblast （HELF）. Results HCMV pp65 antigen was localized in different part of the positive peripheral leukocytes： cytoplasm type, nucleus type, and cytoplasm and nucleus mixed type. In the nucleus type pp65 antigen was inside the cell nucleus. Only coeuhure with HCMV wild strain infected HELF, pp65 positive leukocytes could be deteced, Cytoplasm positive leukncytes just appeared at the early time of cocuhure（within 1 h） , after cocuhure for 2 h, about 80%-90% positive leukocytes were nucleus type, at advanced stage some cytoplasm and nucleus mixed tpye could be found, Conclusions There are three types of HCMV pp65 antigen positive leukocytes in peripheral blood. In the nucleus type, pp65 antigen is not only accretes the surface of nucleus, but also gathers in the cell nucleus. Coculture of leukocyte with HCMV infected HELF shows that only nucleus positive leukocyte indicates HCMV activation.
出处
《检验医学》
CAS
北大核心
2008年第2期131-134,共4页
Laboratory Medicine
基金
上海市教育委员会科研项目(04BA04)
