Qureshi A A, Trent D W
Infect Immun. 1973 Dec;8(6):993-9. doi: 10.1128/iai.8.6.993-999.1973.
Solubilized nonstructural antigen III from Saint Louis encephalitis (SLE)-Japanese B encephalitis (JBE)-West Nile (WN) and dengue-2 arbovirus-infected pig kidney cells was purified by employing Brij-58 solubilization, organic solvent extraction, and column chromatography. Diethylaminoethyl-cellulose column peak C eluate contained only intracellular viral envelope protein designated antigen I. Antigen I of SLE virus absorbed homologous neutralizing antibody; however, the intracellular nonstructural protein designated antigen III did not. The antigenic relationships of solubilized antigens I and III prepared from SLE, JBE, and WN virus-infected cells were determined by complement-fixation (CF) and immunodiffusion analyses. Solubilized antigen I of each virus cross-reacted broadly with both homologous and heterologous antibody at high antigen concentrations in the CF test. Antigen III of each virus reacted with only homologous antibody by both CF and immunoprecipitation. This study demonstrates that during SLE, JBE, WN, and dengue-2 infections, virus-specific proteins containing both type-specific and group-reactive determinants are synthesized. Antigen III, a nonstructural protein, is serologically virus type specific and may be useful as a type-specific diagnostic reagent.
通过采用Brij - 58增溶、有机溶剂萃取和柱色谱法,对来自圣路易斯脑炎(SLE)、日本乙型脑炎(JBE)、西尼罗河(WN)和登革热2型虫媒病毒感染的猪肾细胞中增溶的非结构抗原III进行了纯化。二乙氨基乙基纤维素柱峰C洗脱液仅含有细胞内病毒包膜蛋白,命名为抗原I。SLE病毒的抗原I能吸附同源中和抗体;然而,细胞内的非结构蛋白(命名为抗原III)则不能。通过补体结合(CF)和免疫扩散分析,确定了从SLE、JBE和WN病毒感染细胞中制备的增溶抗原I和III的抗原关系。在CF试验中,每种病毒的增溶抗原I在高抗原浓度下与同源和异源抗体均有广泛的交叉反应。每种病毒的抗原III仅通过CF和免疫沉淀与同源抗体发生反应。本研究表明,在SLE、JBE、WN和登革热2型感染期间,合成了含有型特异性和组反应性决定簇的病毒特异性蛋白。抗原III作为一种非结构蛋白,在血清学上具有病毒型特异性,可用作型特异性诊断试剂。