摘要
采用Ficoll密度梯度离心法(淋巴细胞分离液)分离肾综合征出血热(HFRS)患者外周血单个核细胞(PBMC),并用EB病毒(EBV)感染B淋巴细胞,建立永生化的B淋巴母细胞系(B-LCL)。然后,用含汉滩病毒(Hantaan virus,HTNV)S基因的重组痘苗病毒感染B-LCL,应用间接免疫荧光检测核衣壳蛋白的表达。结果表明,B淋巴细胞经EBV感染4周左右,可形成永生化B-LCL。成功转化后的B-LCL,体积增大,且增殖的淋巴细胞积聚成团。汉滩病毒S基因在B-LCL中能有效表达核衣壳蛋白。含S基因的重组痘苗病毒感染的B-LCL可用作HTNV核衣壳蛋白特异性CTL活性研究的靶细胞。
Peripheral blood mononuclear cells (PBMCs) were separated on Ficoll-Hypaque density gradient centrifugation from patients with Hemorrhagic Fever with Renal Syndrome (HFRS) and infected by EB virus (EBV), in order to establish immortalized B lymphoblastoid cell line (B-LCL). B-LCLs were then infected with the recombinant vaccinia viruse harboring Hantaan virus (HTNV) S gene. Expression of nucleocapsid protein was detected using indirect immunofluorescent assay (IFA). The results showed that immortalized B-LCLs were set up, after B lymphocytes were infected by EBV and cultured for 4 weeks. B lymphocytes transformed by EBV became enlarged and further gathered together. The HTNV nucleocapsid protein can be effectively expressed in B-LCLs transinfected with recom- binant vaccinia viruse harboring HTNV-S gene.
出处
《中国病毒学》
CSCD
2003年第5期415-419,共5页
Virologica Sinica
关键词
汉滩病毒
重组痘苗病毒
肾综合征出血热
EB病毒
B淋巴母细胞系
Hantaan virus (HTNV)
Recombinant vaccinia virus(rVV)
Hemorrhagic Fever with Renal Syndrome (HFRS)
EB virus(EBV), B lymphoblastoid cell lines (B-LCL)
