Dana obtained her BSc (Hons) in Biomedical Science from the University of Surrey in 2018. As part of this programme she undertook a professional training year within the Porcine Reproductive and Respiratory Syndrome (PRRS) Immunology group at The Pirbright Institute, between 2016-2017. She assisted in evaluating a methodology to effectively genetically modify porcine B cells, during which she developed a keen interest in bioimaging techniques.
Dana is currently undertaking her PhD at the University of Surrey, in collaboration with The Pirbright Institute, under the mentorship of Prof. Gill Elliott and Prof. Pippa Hawes. Her project aims to investigate the morphogenesis of the prototypic alphaherpesvirus herpes simplex virus type 1 (HSV-1), utilising recombinant fluorescently-tagged viruses to examine tegument proteins within the infected cell.
The rapid evolution of porcine reproductive and respiratory syndrome viruses (PRRSV) poses a major challenge to effective disease control since available vaccines show variable efficacy against divergent strains. Knowledge of the antigenic targets of virus-neutralizing antibodies that confer protection against heterologous PRRSV strains would be a catalyst for the development of next-generation vaccines. Key to discovering these epitopes is the isolation of neutralizing monoclonal antibodies (mAbs) from immune pigs. To address this need, we sought to establish systems to enable the isolation of PRRSV neutralizing porcine mAbs. We experimentally produced a cohort of immune pigs by sequential challenge infection with four heterologous PRRSV strains spanning PRRSV-1 subtypes and PRRSV species. Whilst priming with PRRSV-1 subtype 1 did not confer full protection against a subsequent infection with a PRRSV-1 subtype 3 strain, animals were protected against a subsequent PRRSV-2 infection. The infection protocol resulted in high serum neutralizing antibody titers against PRRSV-1 Olot/91 and significant neutralization of heterologous PRRSV-1/-2 strains. Enriched memory B cells isolated at the termination of the study were genetically programmed by transduction with a retroviral vector expressing the Bcl-6 transcription factor and the anti-apoptotic Bcl-xL protein, a technology we demonstrated efficiently converts porcine memory B cells into proliferating antibody-secreting cells. Pools of transduced memory B cells were cultured and supernatants containing PRRSV-specific antibodies identified by flow cytometric staining of infected MARC-145 cells and in vitro neutralization of PRRSV-1. Collectively, these data suggest that this experimental system may be further exploited to produce a panel of PRRSV-specific mAbs, which will contribute both to our understanding of the antibody response to PRRSV and allow epitopes to be resolved that may ultimately guide the design of immunogens to induce cross-protective immunity.