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

摘要

Vaccination with autologous cancer cells expressing a potent foreign antigen is promosing 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 form the allantoic fluid of DNA-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 myelona 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.