Antigenic analysis of nonstructural protein (NSP) 4 of group A avian rotavirus strain PO-13.
In order to analyze the antigenic structure of nonstructural protein (NSP) 4 of group A avian rotavirus strain PO-13, 25 monoclonal antibodies (MAbs) against NSP4 expressed in Escherichia coli were produced. All MAbs reacted with NSP4 on Western blotting, indicating that they recognized sequential epitopes. To determine the antigenic sites (ASs) recognized by the produced MAbs, seven truncated NSP4s were expressed in E. coli. Western blotting analysis showed that there are at least four major ASs on PO-13 NSP4, designated as AS I located in amino acids (aa) 151 to 169, AS II (aa 136 to 150), AS III (aa 112 to 133) and AS IV (aa 1 to 24). Two MAbs reacted exclusively with AS III encompassing the region that has been reported to be an enterotoxin domain. MAbs against ASs II, III and IV reacted with all avian rotaviruses tested by indirect immunofluorescent antibody assays. MAbs against AS I reacted with turkey strains, Ty-1 and Ty-3, but not with a chicken strain, Ch-1. Nine of 11 MAbs against AS II cross-reacted with NSP4 of mammalian rotavirus strains with different NSP4 genotypes. These results suggest that AS II on NSP4 is widely conserved among a variety of rotaviruses.
Laboratory of Zoonotic Diseases, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University, Gifu, Gifu 501-1193, Japan.
Mohammed Ali
MA
Borgan
Yoshio
Y
Mori
Naoto
N
Ito
Makoto
M
Sugiyama
Nobuyuki
N
Minamoto
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000956:Antigens, Viral / Q000276:immunology*; D002460:Cell Line; D003001:Cloning, Molecular; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D009154:Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012401:Rotavirus / Q000276:immunology* / Q000302:isolation & purification*; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D011994:Recombinant Proteins; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins