Identification of potential B cell epitope determinants by computer techniques, in hemagglutinin-neuraminidase from the porcine rubulavirus La Piedad Michoacan.
Hemagglutinin-neuraminidase (HN) from porcine rubulavirus La Piedad Michoacan (RvpLPM) is one of the most antigenic proteins known, and is responsible for virus-host cell interaction. We analyzed the amino acid sequence of HN, using computer-assisted techniques to identify B cell epitopes. From a pool of 18 possible antigenic peptides, we evaluated the antigenicity of the 2 peptides with the highest scores and the 1 with lowest score. Antibodies from RvpLPM-infected pigs recognized the synthesized HN-A, HN-B, and HN-R peptides (optical density [OD]: 0.33 +/- 0.02 for HN-A, 0.20 +/- 0.02 for HN-B, and 0.07 +/- 0.01 for HN-R); bovine serum albumin-coupled HN-A and HN-B induced rabbit anti-RvpLPM antibodies (OD: 0.39 +/- 0.01 for HN-A and 0.35 +/- 0.02 for HN-B). Loop 5 from the outer membrane protein, OmpC, from Salmonella typhi was replaced with HN-B; this protein was then expressed in Escherichia coli UH302. BALB/c mice were challenged intraperitoneally or orogastrically with the fusion protein expressed in E. coli and murine antibodies obtained from both types of administration inhibited virus-hemagglutinating activity, as did the antibodies from RvpLPM-infected swine. These results suggest that HN-A and HN-B are peptides involved in RvpLPM cell carbohydrate recognition, and could therefore be considered potential targets for vaccine and diagnostic procedures development.
Zenteno-Cuevas
Roberto
R
Instituto de Salud Pública, Universidad Veracruzana, Xalapa, Veracruz, Mexico. rzenteno@uv.mx
Huerta-Yepez
Sara
S
Reyes-Leyva
Julio
J
Hernández-Jáuregui
Pablo
P
González-Bonilla
Cesar
C
Ramírez-Mendoza
Humberto
H
Agundis
Concepción
C
Zenteno
Edgar
E
D000465:Algorithms; D000818:Animals; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D015707:HN Protein / Q000737:chemistry / Q000276:immunology*; D006385:Hemagglutination Inhibition Tests; D006387:Hemagglutination, Viral; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification; D019213:Rubulavirus / Q000276:immunology*; D019351:Rubulavirus Infections / Q000276:immunology* / Q000821:virology; D012984:Software; D013552:Swine
D018985:Epitopes, B-Lymphocyte; D015707:HN Protein; D010455:Peptides; D011993:Recombinant Fusion Proteins