Abstract
Dendritic cells (DCs) are important mediators of the immune system, residing
in areas close to the external environment. The roles of DCs in the modulation of the
immune system have long been studied in other mammalian species, the extent of
which has evaded the equine model due to a lack of equine specific reagents required
for investigation. An antibody panel was established in order to interrogate the
phenotypes of equine peripheral blood mononuclear cell (PBMC) subsets. Using flow
cytometry, putative pDC, cDC1, cDC2, monocytes and two unknown subsets were
identified and align with those of other mammalian species. EqWC1 was employed
successfully as an addition to the panel and showed clear differential staining patterns
across the subsets, although the identity of the target of this antibody remains
unclear. To overcome limitations posed by the lack of equine antibodies primer panels
were established to provide downstream tools for interrogating the putative DC
populations. To further the characterisation of equine DC, PBMCs were depleted of T
and B cells and interrogated via molecular techniques, such as transcriptome and
single cell RNA sequencing. Thereby, cDC1, cDC2, pDC, and monocytes were
confirmed. Following culture under different cytokine environments, it was evident
that dendritic cells did better and had an increased cell viability when monocytes were
included. In summary, work carried out in this thesis provides fundamental insight to
further study and use of DC in infection and vaccine models.