Expression of the Cloned Hemagglutinin-Neuraminidase Gene of the Newcastle Disease Virus in Mouse Myeloma Cells

Vaccination with autologous cancer cells expressing a potent foreign antigen is promising for immunotherapy of tumors. A construct was obtained to transfect cancer cells with the hemagglutinin-neuraminidase (HN) gene of the Newcastle disease virus (NDV). Specific primers were designed, and the HN cD...

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Bibliographic Details
Published in:Molecular biology (New York) 2001-09, Vol.35 (5), p.785-791
Main Authors: Risinskaya, N V, Fegeding, K V, Vasilenko, O V, Sudarikov, A B
Format: Article
Language:English
Subjects:
Allantoic fluid
Birds
Cancer
Cancer vaccines
Cell agglutination
double prime HN gene
Eggs
Enzyme-linked immunosorbent assay
Exo- alpha -sialidase
Expression vectors
Ice
Immunotherapy
Myeloma
Newcastle disease
Newcastle disease virus
Poultry
Primers
Proteins
RNA
Tumors
Vaccination
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Summary:Vaccination with autologous cancer cells expressing a potent foreign antigen is promising for immunotherapy of tumors. A construct was obtained to transfect cancer cells with the hemagglutinin-neuraminidase (HN) gene of the Newcastle disease virus (NDV). Specific primers were designed, and the HN cDNA was amplified from RNA isolated from the allantoic fluid of NDV-infected embryonated chicken eggs. The amplified fragment was cloned in pCR2.1, sequenced, and recloned in expression vector pCDNA3.1/Zeo(+). The resulting construct was used to transfect mouse myeloma cells SP2/0. Production of HN was checked by ELISA and by a neuraminidase activity assay. Cell agglutination on ice was proposed as a test for surface HN.[PUBLICATION ABSTRACT]
ISSN:0026-8933
1608-3245
DOI:10.1023/A:1012346925499