SELF-ASSEMBLED B19 PARVOVIRUS CAPSIDS, PRODUCED IN A BACULOVIRUS SYSTEM, ARE ANTIGENICALLY AND IMMUNOGENICALLY SIMILAR TO NATIVE VIRIONS

被引:191
作者
KAJIGAYA, S
FUJII, H
FIELD, A
ANDERSON, S
ROSENFELD, S
ANDERSON, LJ
SHIMADA, T
YOUNG, NS
机构
[1] CENT PUBL HLTH LAB,VIRUS REFERENCE LAB,LONDON NW9 5HT,ENGLAND
[2] CTR DIS CONTROL,RESP & ENTEROVIRUS DIS BRANCH,ATLANTA,GA 30333
关键词
5TH DISEASE; TRANSIENT APLASTIC CRISIS; VACCINE; ENZYME IMMUNOASSAY;
D O I
10.1073/pnas.88.11.4646
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 B19 VP1 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.
引用
收藏
页码:4646 / 4650
页数:5
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