Epitope mapping studies with neutralizing and non-neutralizing monoclonal antibodies to the G1 and G2 envelope glycoproteins of Hantaan virus.
Epitopes recognized by three G1-specific and two G2-specific neutralizing monoclonal antibodies to Hantaan virus were mapped by sequence analyses of the complete M genome segments of neutralization escape variant viruses. For each variant, we detected nucleotide sequence substitutions which resulted in a single amino acid change in either the G1 or G2 protein. Serological properties of the variant viruses correlated with changes identified by nucleotide sequence analyses. To map epitopes recognized by three G1-specific and six G2-specific, non-neutralizing monoclonal antibodies, we prepared genes, truncated at the carboxy terminal coding regions of G1 or G2, and expressed them with baculovirus recombinants or transiently in a vaccinia/T7 RNA polymerase system. Reactivities of the monoclonal antibodies with the truncated proteins were monitored by immune precipitation of the radiolabeled, truncated glycoproteins. We determined that all three of the G1-specific antibodies reacted with truncated proteins, which retained the amino terminal one-third of G1, but lost reactivity with shorter G1 proteins. The G2-specific antibodies only recognized G2 proteins, which retained approximately 80% of the G2 gene.
Wang
M
M
Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21702-5011.
Pennock
D G
DG
Spik
K W
KW
Schmaljohn
C S
CS
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000276:immunology*; D016367:Baculoviridae / Q000235:genetics; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005814:Genes, Viral; D014644:Genetic Variation; D018063:Hantaan virus / Q000235:genetics / Q000276:immunology*; D006385:Hemagglutination Inhibition Tests; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000276:immunology; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins