A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein | Herrera et al. Virology Journal 2010 7 195 http content 7 1 195 VIROLOGY JOURNAL RESEARCH Open Access A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein 11 11 1 2 3 Emma Herrera11 Patricia Barcenas Rubicela Hernandez Alfonso Mendez Guillermo Pérez-Ishiwara Blanca Baiif-n Abstract Background The hemagglutinin-neuraminidase HN protein is the major antigenic determinant of the Mumps virus MuV and plays an important role in the viral infectious cycle through its hemagglutination hemadsorption HA HD and neuraminidase NA activities. Objective analyze the biological and immunological properties of a polypeptide derived from a highly conserved region of the HN ectodomain. Methods a highly conserved region of the HN gene among several MuV genotypes was chosen to be cloned in a eukaryotic expression vector. The pcDNAHN176-construct was transfected into Vero cells and RNA expression was detected by RT-PCR while the corresponding polypeptide was detected by immunofluorescence and immunochemistry techniques. The HD and NA activities were also measured. The immunogenic properties of the construct were evaluated using two systems rabbit immunization to obtain sera for detection of the HN protein and neutralization of MuV infection and hamster immunization to evaluate protection against MuV infection. Results A 567 nucleotide region from the HN gene was amplified and cloned into the plasmid . Vero cells transfected with the construct expressed a polypeptide that was recognized by a MuV-hyperimmune serum. The construct-transfected cells showed HD and NA activities. Sera from immunized rabbits in vitro neutralized two different MuV genotypes and also detected both the HN protein and the HN176 polypeptide by western blot. Hamsters immunized with the pcDNAHN176-construct and challenged with MuV showed a mild viral infection in comparison to non-immunized animals and .
