Mapping of B-cell epitopes of hemagglutinin protein of rinderpest virus.
Monoclonal antibodies (mAbs) against secreted hemagglutinin (H) protein of rinderpest virus (RPV) expressed by a recombinant baculovirus were generated to characterize the antigenic sites on H protein and regions of functional significance. Three of the mAbs displayed hemagglutination inhibition activity and these mAbs were unable to neutralize virus infectivity. Western immunoblot analysis of overlapping deletion mutants indicated that three mAbs recognize antigenic regions at the extreme carboxy terminus (between amino acids 569 and 609) and the fourth mAb between amino acids 512 and 568. Using synthetic peptides, aa 569-577 and 575-583 were identified as the epitopes for E2G4 and D2F4, respectively. The epitopic domains of A12A9 and E2B6 mAbs were mapped to regions encompassing aa 527-554 and 588-609. Two epitopes spanning the extreme carboxy terminal region of aa 573 to 587 and 588 to 609 were shown to be immunodominant employing a competitive ELISA with polyclonal sera form vaccinated cattle. The D2F4 mAb which recognizes a unique epitope on RPV-H is not present on the closely related peste des petits ruminant virus HN protein and this mAb could serve as a tool in the seromonitoring program after rinderpest vaccination.
Project Directorate on Animal Disease Monitoring and Surveillance, Hebbal, Bangalore- 560 024, India.
G J
GJ
Renukaradhya
S
S
Mitra-Kaushik
G
G
Sinnathamby
M
M
Rajasekhar
M S
MS
Shaila
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D002417:Cattle; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D017353:Gene Deletion; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D006389:Hemagglutinins, Viral; D007106:Immune Sera; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012302:Rinderpest virus / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D007106:Immune Sera; D010455:Peptides; D011994:Recombinant Proteins; D014764:Viral Proteins; C079749:hemagglutinin protein, Rinderpest virus