Expression, processing, and assembly of foot-and-mouth disease virus capsid structures in heterologous systems: induction of a neutralizing antibody response in guinea pigs.
Plasmids containing the foot-and-mouth disease virus structural protein precursor (P1) and 3C protease genes or the P1 gene alone were expressed in Escherichia coli. A recombinant baculovirus containing the P1 gene was also generated and expressed in Spodoptera frugiperda cells. Expression of the P1 and 3C genes in E. coli resulted in efficient synthesis and processing of the structural protein precursor and assembly into 70S empty capsids. This material reacted with neutralizing monoclonal antibodies which recognize only conformational epitopes and elicited a significant neutralizing antibody response in vaccinated guinea pigs. Expression of the P1 gene in E. coli resulted in synthesis of an insoluble product, whereas in insect cells infected with the recombinant baculovirus a soluble product was synthesized. Both soluble and insoluble P1 reacted with a 12S-specific monoclonal antibody, but only soluble P1 elicited a neutralizing antibody response in guinea pigs.
Lewis
S A
SA
Plum Island Animal Disease Center, U.S. Department of Agriculture, Greenport, New York 11944.
Morgan
D O
DO
Grubman
M J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D005537:Aphthovirus / Q000235:genetics* / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006168:Guinea Pigs; D009036:Moths; D009500:Neutralization Tests; D010957:Plasmids; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D014162:Transfection
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D011994:Recombinant Proteins