A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein
<p>Abstract</p> <p>Background</p> <p>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 (...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-08-01
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| Series: | Virology Journal |
| Online Access: | http://www.virologyj.com/content/7/1/195 |
| _version_ | 1818037761070858240 |
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| author | Herrera Emma Barcenas Patricia Hernández Rubicela Méndez Alfonso Pérez-Ishiwara Guillermo Barrón Blanca |
| author_facet | Herrera Emma Barcenas Patricia Hernández Rubicela Méndez Alfonso Pérez-Ishiwara Guillermo Barrón Blanca |
| author_sort | Herrera Emma |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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. <it>Objective</it>: analyze the biological and immunological properties of a polypeptide derived from a highly conserved region of the HN ectodomain. <it>Methods</it>: 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.</p> <p>Results</p> <p>A 567 nucleotide region from the HN gene was amplified and cloned into the plasmid pcDNA3.1. 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 <it>in vitro </it>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 Th1 and Th2 cytokines were detected in them.</p> <p>Conclusions</p> <p>The pcDNAHN176-construct was capable of expressing a polypeptide in Vero cells that was identified by a hyperimmune serum anti Mumps virus, and these cells showed the HD and NA activities of the complete MuV HN protein. The construct also elicited a specific immune response against MuV infection in hamsters.</p> |
| first_indexed | 2024-12-10T07:31:58Z |
| format | Article |
| id | doaj.art-a1a36a6976684e1cb452945b6cee196d |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-10T07:31:58Z |
| publishDate | 2010-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-a1a36a6976684e1cb452945b6cee196d2022-12-22T01:57:32ZengBMCVirology Journal1743-422X2010-08-017119510.1186/1743-422X-7-195A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN proteinHerrera EmmaBarcenas PatriciaHernández RubicelaMéndez AlfonsoPérez-Ishiwara GuillermoBarrón Blanca<p>Abstract</p> <p>Background</p> <p>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. <it>Objective</it>: analyze the biological and immunological properties of a polypeptide derived from a highly conserved region of the HN ectodomain. <it>Methods</it>: 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.</p> <p>Results</p> <p>A 567 nucleotide region from the HN gene was amplified and cloned into the plasmid pcDNA3.1. 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 <it>in vitro </it>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 Th1 and Th2 cytokines were detected in them.</p> <p>Conclusions</p> <p>The pcDNAHN176-construct was capable of expressing a polypeptide in Vero cells that was identified by a hyperimmune serum anti Mumps virus, and these cells showed the HD and NA activities of the complete MuV HN protein. The construct also elicited a specific immune response against MuV infection in hamsters.</p>http://www.virologyj.com/content/7/1/195 |
| spellingShingle | Herrera Emma Barcenas Patricia Hernández Rubicela Méndez Alfonso Pérez-Ishiwara Guillermo Barrón Blanca A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein Virology Journal |
| title | A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein |
| title_full | A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein |
| title_fullStr | A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein |
| title_full_unstemmed | A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein |
| title_short | A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein |
| title_sort | 176 amino acid polypeptide derived from the mumps virus hn ectodomain shows immunological and biological properties similar to the hn protein |
| url | http://www.virologyj.com/content/7/1/195 |
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