title: Cloning, expression and characterization of potential immunogenic recombinant hemagglutinin-neuraminidase protein of Porcine rubulavirus.
abstract: Blue eye disease caused by Porcine rubulavirus (PorPV) is an endemic viral infection of swine causing neurological and respiratory disease in piglets, and reproductive failure in sows and boars. The hemagglutinin-neuraminidase (HN) glycoprotein of PorPV is the most abundant component in the viral envelope and the main target of the immune response in infected animals. In this study, we expressed the HN-PorPV-recombinant (rHN-PorPV) protein in an Escherichia coli system and analyzed the immune responses in mice. The HN gene was cloned from the reference strain PorPV-La Piedad Michoacan Virus (GenBank accession number BK005918), into the pDual expression vector. The expressed protein was identified at a molecular weight of 61.7 kDa. Three-dimensional modeling showed that the main conformational and functional domains of the rHN-PorPV protein were preserved. The antigenicity of the expressed protein was confirmed by Western blot with a monoclonal antibody recognizing the HN, and by testing against serum samples from pigs experimentally infected with PorPV. The immunogenicity of the rHN-PorPV protein was tested by inoculation of BALB/c mice with AbISCO-100(®) as adjuvant. Analysis of the humoral immune responses in mice showed an increased level of specific antibodies 14 days after the first immunization, compared to the control group (P < 0.0005). The results show the ability of the rHN-PorPV protein to induce an antibody response in mice. Due to its immunogenic potential, the rHN-PorPV protein will be further evaluated in pig trials for its suitability for prevention and control of blue eye disease.
authors: Centro Nacional de Investigación Disciplinaria en Microbiología Animal, INIFAP, CP. 05110, Mexico City, Mexico; Division of Virology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, S-751 89, Uppsala, Sweden. Electronic address: cuevas.julieta@inifap.gob.mx.
authors: Julieta Sandra
authors: JS
authors: Cuevas-Romero
authors: Centro Nacional de Investigación Disciplinaria en Microbiología Animal, INIFAP, CP. 05110, Mexico City, Mexico.
authors: José Francisco
authors: JF
authors: Rivera-Benítez
authors: Facultad de Estudios Superiores Cuautitlán FES-C-UNAM, Edo. de México, Mexico.
authors: Eliseo
authors: E
authors: Hernández-Baumgarten
authors: Private Researcher, Puebla, Mexico.
authors: Pablo
authors: P
authors: Hernández-Jaúregui
authors: Instituto Politécnico Nacional. Centro de Investigación y Estudios Avanzados, Mexico City, Mexico.
authors: Marco
authors: M
authors: Vega
authors: Division of Virology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, S-751 89, Uppsala, Sweden.
authors: Anne-Lie
authors: AL
authors: Blomström
authors: Division of Virology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, S-751 89, Uppsala, Sweden.
authors: Mikael
authors: M
authors: Berg
authors: Division of Virology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, S-751 89, Uppsala, Sweden.
authors: Claudia
authors: C
authors: Baule
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli; D005260:Female; D015870:Gene Expression; D015707:HN Protein / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D000071497:Immunogenicity, Vaccine; D051379:Mice; D008807:Mice, Inbred BALB C; D019213:Rubulavirus / Q000201:enzymology / Q000276:immunology; D013552:Swine; D014765:Viral Vaccines / Q000096:biosynthesis / Q000276:immunology
chemical_list: D015707:HN Protein; D014765:Viral Vaccines
keywords: Blue eye disease; Gene expression; Hemagglutinin-neuraminidase; Porcine rubulavirus; Recombinant protein