Estes M K, Crawford S E, Penaranda M E, Petrie B L, Burns J W, Chan W K, Ericson B, Smith G E, Summers M D
J Virol. 1987 May;61(5):1488-94. doi: 10.1128/JVI.61.5.1488-1494.1987.
Rotaviruses are the major pathogens that cause life-threatening diarrhea in young children and animals. We inserted a simian rotavirus SA11 gene 6 cDNA into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. The major capsid antigen (VP6) was expressed in high yields (20 to 150 micrograms/10(6) cells) when Spodoptera frugiperda cells were infected with baculovirus recombinants containing SA11 gene 6 inserts. Reactivity with monospecific polyclonal and monoclonal antibodies suggested that VP6, expressed intracellularly or found in the media, maintained native antigenic determinants. VP6 purified from the media from infected cells also possessed a native oligomeric structure, was immunogenic in guinea pigs, and was able to spontaneously assemble into morphologic subunits. Antisera from immunized guinea pigs failed to neutralize virus in plaque reduction assays, but detected homologous and heterologous rotavirus strains when tested by immunofluorescence, immunoprecipitation, and enzyme-linked immunosorbent assays.
轮状病毒是导致幼儿和动物出现危及生命的腹泻的主要病原体。我们将猿猴轮状病毒SA11基因6的cDNA插入到苜蓿银纹夜蛾核型多角体病毒的基因组中，该基因位于强多角体蛋白启动子附近。当用含有SA11基因6插入片段的杆状病毒重组体感染草地贪夜蛾细胞时，主要衣壳抗原（VP6）得以高产表达（20至150微克/10⁶个细胞）。与单特异性多克隆抗体和单克隆抗体的反应表明，在细胞内表达或在培养基中发现的VP6保留了天然抗原决定簇。从感染细胞的培养基中纯化得到的VP6也具有天然的寡聚结构，在豚鼠中具有免疫原性，并且能够自发组装成形态亚基。免疫豚鼠的抗血清在蚀斑减少试验中未能中和病毒，但在通过免疫荧光、免疫沉淀和酶联免疫吸附试验检测时，能够检测到同源和异源轮状病毒株。
