{"id":120,"date":"2020-07-19T00:24:19","date_gmt":"2020-07-18T23:24:19","guid":{"rendered":"http:\/\/blogs.kent.ac.uk\/mspvpuprepare\/?p=120"},"modified":"2020-07-19T00:26:44","modified_gmt":"2020-07-18T23:26:44","slug":"influenza-d-pseudotyped-lentiviruses-production-neutralisation-assay-and-serological-surveillance","status":"publish","type":"post","link":"https:\/\/blogs.kent.ac.uk\/mspvpuprepare\/2020\/07\/19\/influenza-d-pseudotyped-lentiviruses-production-neutralisation-assay-and-serological-surveillance\/","title":{"rendered":"Influenza D pseudotyped lentiviruses: production, neutralisation assay and serological surveillance"},"content":{"rendered":"
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Influenza D virus (IDV) has been reported in many animal species and potentially humans worldwide. Cattle are considered the major reservoir. There are currently three main lineages based on the haemagglutinin-esterase (HEF) gene: D\/OK, D\/660 and D\/Japan. We performed pilot surveillance for IDV by using pseudotyped lentivirus (PVs) to generate a cell-based test to identify prior-exposure to IDV in animals. The expression plasmids of the HEF genes, D\/swine\/Italy\/2015, D\/bovine\/France\/2014, and D\/bovine\/Ibaraki\/2016, were constructed. The HEF plasmid was co-transfected with lentiviral vector plasmid expressing luciferase, lentiviral Gag-Pol plasmid, and HAT protease plasmid in producer cells (HEK293T\/17). Three days post-transfection, supernatants were collected and used for titration on various cell lines and in micro-neutralisation tests. Sera from pigs vaccinated with D\/swine\/Italy\/2015 and D\/swine\/Oklahoma\/2011 were used to undertake a preliminary validation of the micro-neutralisation assay. All pig sera have neutralising activity to influenza D (Italy) pseudotyped lentiviruses. Cow and sheep sera, 145 and 114 specimens, respectively, collected from UK farms were screened using the micro-neutralisation test. We found 97 bovine sera (66.9%) were influenza D antibody positive. Collectively, pseudotyped lentivirus technology opens up opportunities for serological surveillance of influenza D viruses.<\/p>\n<\/div>\n<\/div>\n<\/div>\n