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Name Isobel Braithwaite Home University University of Cambridge Broad subject area of Amgen Scholars project Infection and

Immunity Supervisor(s) Antonio Barragan Supervisor(s) department(s) KI Parasitology, Microbiology and Virology/ Swedish Centre for Transmissible Disease Control Poster title Characterisation of changes in DC phenotype induced by T. gondii and investigation of the molecular mechanisms involved Authors Isobel Braithwaite, Jess Weidner Abstract (350 words maximum) Toxoplasma gondii is an obligate intracellular parasite estimated to infect at least one third of the global population, although only a tiny fraction cause severe pathology, mainly congenitally and in immunocompromised hosts. T. gondii has been demonstrated to induce a dramatic increase in motility in dendritic cells; professional antigen presenting cells and key regulators of the immune response. This hijack of host cell motility occurs early after cell invasion and has been proposed to have evolved to facilitate parasite dissemination to immune privileged sites and so aid the establishment of latency the Trojan Horse hypothesis. Furthermore, T. gondii has been shown to induce these changes without protein synthesis but only when it is allowed to actively invade. There are also clear morphological changes associated with DC maturation loss of podosomes, appearance of veils and blebbing and taking on a more rounded shape - even at very early time points. This was investigated by transmigration assay, in which the fraction of cells passing through fine pores was determined, alongside a motility assay developed by the group which uses real time imaging and provides more detailed information about cell movement, and showed a. In parallel with this, infected and non-infected human DCs were imaged by confocal microscopy after phalloidin staining. We are currently starting to investigate the protein responsible for the phenotype change through fractionation of intracellular components, separation by ion exchange and size exclusion chromatography, and mass spectrometry, using the assays described above on transfected DCs to look at fraction activity.

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