Li B, Hoshino Y, Gorziglia M
Epidemiology Section, National Institute of Allergy and Infectious Diseases, Bethesda, MD, USA.
Arch Virol. 1996;141(1):155-60. doi: 10.1007/BF01718596.
A cDNA clone representing the VP4-encoding gene of human rotavirus strain 69M(VP7 serotype 8) was constructed and inserted into a baculovirus expression vector. Baculovirus recombinants that expressed the 69 M VP4 protein in Spodoptera frugiperda (Sf9) cells were screened by immunofluorescence with hyperimmune antiserum to the 69M strain and purified by terminal dilution. The expressed VP4 was detected by Coomassie blue staining of PAGE-separated proteins. The antigenic relationships between the VP4 of the 69M strain and those of various human and other animal rotavirus strains representing ten established VP4 serotypes were examined by plaque reduction neutralization. Hyperimmune antiserum produced in guinea pigs following immunization with a lysate of Sf9 cells infected with a 69M gene 4-baculovirus recombinant neutralized the infectivity of the homologous human rotavirus 69M strain as well as heterologous equine rotavirus H-2 strain to a high titer. The anti-69M VP4 hyperimmune antiserum did not neutralize significantly other rotavirus strains of human, simian, porcine, bovine, or murine origin. It thus appears that the human rotavirus 69M strain has a distinct VP4 serotype (designated as P serotype 4) which is closely related antigenically to equine rotavirus H-2 VP4.
构建了一个代表人类轮状病毒69M株（VP7血清型8）编码VP4基因的cDNA克隆，并将其插入杆状病毒表达载体中。通过用针对69M株的超免疫抗血清进行免疫荧光筛选，在草地贪夜蛾（Sf9）细胞中表达69M VP4蛋白的杆状病毒重组体，并通过终点稀释法进行纯化。通过对PAGE分离的蛋白质进行考马斯亮蓝染色来检测表达的VP4。通过蚀斑减少中和试验检测69M株的VP4与代表十种已确立的VP4血清型的各种人类和其他动物轮状病毒株的VP4之间的抗原关系。用感染了69M基因4杆状病毒重组体的Sf9细胞裂解物免疫豚鼠产生的超免疫抗血清，能高效中和同源人类轮状病毒69M株以及异源马轮状病毒H - 2株的感染性。抗69M VP4超免疫抗血清对其他人类、猿猴、猪、牛或鼠源的轮状病毒株没有明显的中和作用。因此，人类轮状病毒69M株似乎具有一种独特的VP4血清型（命名为P血清型4），其在抗原上与马轮状病毒H - 2 VP4密切相关。
