马立克氏病病毒MEQ蛋白单克隆抗体的制备及应用研究

基因表达与病毒感染细胞及致瘤能力之间关系的阐明，将有助于我们对马立克氏病病毒（MDV）meq基因功能的了解，该研究利用通过杆状病毒载体在昆虫细胞系SF9上高度表达的MEQ蛋白免疫BALB/c小鼠，然后收获其免疫脾细胞与肿瘤细胞系SP2/0通过PEG于体外融合，获得的杂交瘤细胞被克隆并通过与MDV感染的鸡胚成纤维细胞（CEF）做免疫荧光试验（FA），进行其分泌抗MEQ单克隆抗体（McAb）能力的筛选，获得4株稳定产生抗MEQ蛋白McAb的杂交瘤细胞，其中3G12E6通过FA和免疫酶组化技术能够检测到MDV致瘤株感染的CEF及自然MD肿瘤细胞中表达的meq基因产物。研究的结果首次发现，MDV致瘤株GA、RB1B感染的CEF及自然MD肿瘤细胞中均有meq基因的表达，但在弱毒疫苗株CV1988感染的CEF和免疫鸡组织细胞中则未检测到MEQ，作者认为这是由于meq基因在致瘤株中表达水平高，而在非致瘤株中表达水平低所致，这可能是决定MDV毒力（virulence）或致瘤性（oncogenicity）的关键因素之一。

The development of monoclonal antibody(McAb) against MEQ protein of Marek''''s disease virus(MDV) and its applications

The revelation of the relationship between the expression of gene and the ability of infection, oncogenesis of MDV in vitro and in vivo may play a role in the function analysis of MDV meq gene. In the study, the MEQ protein highly expressed in the insect cell line SF9 by the baculovirus vector was immunized into BALB/c mice and the immunized spleen cells were collected and fused with the tumor cell line SP2/0 via PEG1000 in vitro . The hybridoma cells were cloned and screened for the ability of antiMEQ McAb secretion by immunofluorescence assay (FA) with the MDV GA infected chicken embryo fibroblast (CEF). Four McAbs were developed and can detect the meq gene expression in the CEF infected with oncogenic MDVs and naturally occurred MD tumor cells by FA and immunohistochemistry technique respectively, but can't detect that in the CEF infected with nonvirulent vaccine virus CVI988/Rispens. The results demonstrated that meq is highly expressed in the oncogenic MDVs infected CEF in vitro and the naturally occurred MD tumor cells in vivo when compare to that of the nonvirulent vaccine virus and that consequently may play an important role in the virulence or oncogenicity of MDV.