Kajigaya S, Fujii H, Field A, Anderson S, Rosenfeld S, Anderson L J, Shimada T, Young N S
Cell Biology Section, National Heart, Lung, and Blood Institute, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4646-50. doi: 10.1073/pnas.88.11.4646.
B19 parvovirus is pathogenic in humans, causing fifth disease, transient aplastic crisis, some cases of hydrops fetalis, and acquired pure red cell aplasia. Efforts to develop serologic assays and vaccine development have been hampered by the virus's extreme tropism for human bone marrow and the absence of a convenient culture system. We constructed recombinants containing either the major (VP2) or minor (VP1) structural proteins of B19 in a baculovirus-based plasmid, from which the polyhedrin gene had been deleted; these recombinant plasmids were used to generate recombinant infectious baculovirus. Subsequent infection of insect cells in vitro resulted in high-level expression of either B19VP1 or VP2. Parvovirus capsids were obtained by self-assembly in cell cultures coinfected with either VP1- and VP2-containing baculoviruses or, surprisingly, VP2-containing baculoviruses alone. Empty B19 capsids composed of VP1 and VP2 could replace serum virus as a source of antigen in a conventional immunoassay for detection of either IgG or IgM antiparvovirus antibodies in human serum. Immunization of rabbits with capsids composed of VP1 and VP2 resulted in production of antisera that recognized serum parvovirus on immunoblot and neutralized parvovirus infectivity for human erythroid progenitor cells. Baculovirus-derived parvovirus antigen can substitute for scarce viral antigen in immunoassays and should be suitable as a human vaccine.
B19细小病毒对人类具有致病性,可引发传染性红斑、再生障碍性危象、部分胎儿水肿病例以及获得性纯红细胞再生障碍。由于该病毒对人类骨髓具有极强的嗜性,且缺乏便捷的培养系统,血清学检测方法的开发和疫苗研发工作受到了阻碍。我们构建了重组体,将B19的主要结构蛋白(VP2)或次要结构蛋白(VP1)置于一个缺失了多角体蛋白基因的杆状病毒质粒中;这些重组质粒用于产生重组感染性杆状病毒。随后在体外感染昆虫细胞,导致B19VP1或VP2高水平表达。细小病毒衣壳通过在同时感染含VP1和VP2的杆状病毒或单独感染含VP2的杆状病毒的细胞培养物中自组装获得。由VP1和VP2组成的空B19衣壳可替代血清病毒,作为传统免疫测定中检测人血清中抗细小病毒IgG或IgM抗体的抗原来源。用由VP1和VP2组成的衣壳免疫兔子,可产生能在免疫印迹上识别血清细小病毒并中和细小病毒对人红系祖细胞感染性的抗血清。杆状病毒衍生的细小病毒抗原可在免疫测定中替代稀缺的病毒抗原,且应适合作为人用疫苗。