CASE: Generating epithelial differentiation models to explore impact of oral mucosal immunity on EBV replication
University of Registration: University of Warwick
Non-academic partner: Dr Elliot Bland, isoBio
Project Outline
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus1 associated with the development of both lymphoid and epithelial tumours; and most recently, implicated in the development of autoimmune diseases such as MS, ME and long covid2. A key challenge in the study of EBV in epithelial cells is that in vitro models are not well developed, with the more advanced models being developed in B cells3.
This project will aim to develop cutting edge models of epithelial differentiation and organotypic raft cultures for the study of EBV life cycle. These models are well defined for other purposes but have not been established in the context of EBV. This will involve first setting up the model systems and then validating which models allow for EBV replication. Techniques will include 3D culturing of cells, differentiating stem cells, RNAscope and RNASeq for example.
More complex in vitro models of EBV life cycle will be a key starting point in determining virus ability to reactivate in tissues and systems to measure therapeutic and immunological blocking of viral replication. Therefore, following model validation, the models will be used as a platform to test for IgG (systemic immunity) vs IgA (mucosal immunity) ability to halt viral replication. Resulting in an interesting comparison between antibody isotypes and therefore systemic and mucosal immunity.
This project includes collaboration with iosBio, a vaccine company developing oral vaccine against EBV. Antibodies tested will be generated from lead vaccine candidates from this company.
Objectives
1. Develop organotypic raft models of epithelial differentiation
2. Monitor EBV replication in these models
3. Fully characterise models that allow for EBV replication
4. Test antibody isotype and their ability to prevent EBV replication
References
1. Bridgewater, H. E., et al. (2020). The Epstein-Barr Virus-Encoded EBNA1 Protein Activates the Bone Morphogenic Protein (BMP) Signalling Pathway to Promote Carcinoma Cell Migration. Pathogens, 9(7), 594. https://doi.org/10.3390/pathogens9070594.
2. Young, L. S., Yap, L. F., & Murray, P. G. (2016). Epstein-Barr virus: more than 50 years old and still providing surprises. Nature reviews. Cancer, 16(12), 789–802. https://doi.org/10.1038/nrc.2016.92.
3. Bierbaumer, L., Schwarze, U. Y., Gruber, R., & Neuhaus, W. (2018). Cell culture models of oral mucosal barriers: A review with a focus on applications, culture conditions and barrier properties. Tissue barriers, 6(3), 1479568. https://doi.org/10.1080/21688370.2018.1479568.
Candidates are encouraged to contact Dr Hannah Bridgewater to discuss the project before applying if they wish to.
Application
Deadline: 16 January 2025
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