Periodic Report Summary 1 - MAJARVISCIG (A Single Dose, Cytomegalovirus-based Vaccine to Induce Heterosubtypic Protective Immunity to Influenza A Virus)
Our project uses a novel immunogenic vector called cytomegalovirus (CMV), which is biased towards induction of cytotoxic T lymphocytes (CTLs), as a means to induce cross-reactive (heterosubtypic) immunity against the 18 possible influenza A virus (IA) serotypes (H1 through H18, based on hemagglutinin (HA)). IA 'pandemics' with high mortality result from introduction of IA with a new surface HA serotype into an immunologically naïve population. Currently there is no commercial vaccine for pandemic IA. We hypothesis that by using CMV to focus the CTL response on more conserved internal regions of IA, the provided heterosubtypic immunity will protect against pandemic IA. We have used state-of-the-art bacterial artificial chromosome (BAC)-based technology to construct a panel of murine CMV (MCMV) recombinants each expressing a different region of conserved internal IA proteins known to induce CTLs (called a T cell epitope) in mice. These CTL epitopes are derived from the conserved internal IA proteins: nucleoprotein (NP), polymerase acidic protein (PA) – a subunit of the IA polymerase complex, and the nonstructural protein 2 (NS2), a protein involved in nuclear export. The MCMV-IA recombinants express IA H1 versions of these NP, PA and NS2 CTL epitopes fused to a non-essential protein of MCMV called IE2. Fusion of CTL epitopes from other viruses to MCMV IE2 has been shown to induce substantial (>1%) CTL cells against the targeted epitope. This panel of MCMV-IA recombinants will be used to define immunogenicity and efficacy induced by these vectors in the C57BL/6 flu mouse model. Two independent clones of each MCMV-IA recombinant were constructed (MCMV-IANP, MCMV-IAPA and MCMV-IANS2). Restriction digestion combined with gel electrophoresis confirmed the absence of gross genomic rearrangements of the MCMV genome during BAC recombination. Direct DNA sequence analysis confirmed integrity of fusion between the IA epitope and the IE2 protein within the IE2 gene, and growth analysis of reconstituted MCMV-IA viruses in permissive murine fibroblast cells showed virus growth kinetics comparable to wild type (WT) MCMV. Virus stocks were produced for in vivo studies. Two immunogenicity and efficacy studies have been performed in collaboration with our UK and US partners: one against low pathogenic PR8 homotypic (H1N1) IA and the other against highly pathogenic avian (H5N1) IA. The results of these studies are undergoing final analysis. In terms of career integration, the CIG Fellow’s Research Excellence Framework (REF) submission was the second highest for the University of Plymouth. He became a Fellow of the Higher Education Academy (FHEA) following completion of the PGCAP in 2014, for which he was also awarded the PGCAP 2013/2014 Year Cohort Award of Excellence for Innovation in Teaching. In April of 2014, he was promoted from Lecturer to Reader in Virology and Immunology and also appointed Director of Postgraduate Studies in the School. The CIG Fellow has been active in outreach, which was acknowledged by being awarded Marie Curie Fellow of the week in early 2014 for one outreach program in school children (see: www.thejarvislab.com).
Neil Avent, (Head of School of Biomedical & Biological Sciences)
Record Number: 182579 / Last updated on: 2016-05-19
Information source: SESAM