肾综合征出血热病毒对人血管内皮细胞感染性的研究  

Human endothelial cells infected by HFRS virus

作　　者：江振友[1] 温筱芸[1] 林晨[1] 韦静[1] 

机构地区：[1]暨南大学医学院微生物学与免疫学教研室,广州510632

出　　处：《中国人兽共患病杂志》2003年第3期59-62,88,共5页Chinese Journal of Zoonoses

基　　金：广东省医学科研基金资助 (NoA19992 80 )

摘　　要：目的　研究肾综合征出血热 (HFRS)病毒在血管内皮细胞 (HEC)内的增殖 ,及病毒对HEC产生几种细胞因子的影响。方法　采用微量免疫酶斑法检测HFRS病毒A6 9株感染HEC后病毒繁殖动态变化 ,间接免疫荧光法检测HEC内HFRS病毒抗原。ELISA法检测HFRS病毒感染上清液中TNF -α、IL - 6和IFN - β的含量。 结果　病毒感染后 1d即可在培养上清中用微量免疫酶斑法测出病毒 ,病毒滴度 5d达高峰 ,以后下降。在一定范围内病毒产量随病毒感染复数 (multiplicityofinfection ,M0I)的增加而增高。间接免疫荧光法证明感染的HEC胞浆及胞膜上携带肾综合征出血热病毒抗原。电镜和光镜下 ,感染细胞未见明显的形态和结构改变。病毒对HEC产生TNF -α无影响 ;产生IL - 6和IFN - β明显增高。 结论　HFRS病毒可在原始靶细胞HEC内增殖。HFRS病毒感染后所出现的微血管损害可能不是病毒直接损害所致 ,而很可能与间接因素有关。Aim To study the multiplication at hemorrhagic fever with renal syndrome (HFRS) virus in human umbilical vein endothelial cells(HUVEC) and to study the effect of HFRS virus infection on the production of interleukin 6(IL 6),tumor necrosis factor alpha (TNF α) and beta inaerferon(IFN β) secreted by HUVEC Methods HFRS virus pathogenic strain (A69) infected HEC in vitro,culture supernatants were collected at different time postinfection (1d?3d?5d?7d?9d),the titers of virus were measured by microimmunoenzymatic plaque method HFRS virus antigen in human endothelial cell were demonstrated by an indirect immunofluorescent assay The levels of TNF α, IL 6 and IFN β in culture supernatants were measured by ELISA Results The viruses were detected in the supematant as early as day 1 postinfection by microimmunoenzymatic plaque method The highest viral titers were obtained at 5 days,decreasing yields of virus showed at later time(days 4 to 9) Viral production of infeted cells was depended on the multiplicity of infection (MOI) As the viral dose was increased from MOI of 0 1EPFU/cell to 50EPFU/celI We were able to detect the presence of viral antigens in infected HEC by indirect immunofluorescence as early as day 1 postinfection No obvious differences in morphology and structure were observed between the virus infected cells and uninfected cells under phase contrast microscope and transmission electron microscope IFN β and IL 6 production by HUVEC were increased with HFRS virus infection No significant changes were observed in the levels of TNF α Conclusion HEC as HFRS virus original target cell can produce infective virus The mechanism of microvascular damage is not likely to be a direct effect of virus replication but may conceivably be the consequence of an immune mediated endothelial injury triggered by viral infection The production of cytokines may play an important role in HFRS virus pathogenicity and immunity

关 键 词：肾综合征出血热病毒 人血管内皮细胞感染性 研究 微量免疫酶斑法 

分 类 号：R512.8[医药卫生—内科学]