AbstractsBiology & Animal Science

Despite the remarkable success of vaccination, there are a range of human and veterinary diseases that are in need of new vaccines. The rational design of vaccines against these diseases relies on increased understanding of the immunological mechanisms that contribute to vaccine immunity, the development of novel vaccine delivery systems and the characterisation of immune stimulants that are able to increase vaccine efficacy. Vaccine formulations incorporating stimulants that target innate immune receptors have been shown to significantly increase vaccine induced immunity. When incorporated into liposome-based delivery systems, the TLR ligands CpG and poly(I:C) are able to induce protective, long lasting cellular and humoral immune responses in mice. However, the cellular targets of these liposomal adjuvant formulations and the in vivo mechanisms of immune induction remain to be elucidated. The early immune response to vaccination is characterised by activation of cells present at the injection site and their subsequent migration to the local lymph node via the afferent lymphatics. The immunological signals received by innate cells at the peripheral injection site are conveyed to lymphocytes in the local lymph node, leading to the generation of an adaptive immune response where antigen specific lymphocytes emigrate via the efferent lymphatics to perform their tailored effector function. Examination of the afferent and efferent lymphatic compartments during the innate and adaptive phases of an immune response permits the quantification and characterisation of the in vivo biological mechanisms triggered following vaccination. By directly cannulating the ovine lymphatic vessels, the results of this thesis demonstrate that the addition of poly(I:C) or CpG to a liposomal vaccine formulation enhances the immediate inflammatory response at the site of injection, improves antigen uptake by innate cell populations and induces genetic signatures associated with interferon-mediated antiviral immune responses in afferent lymph. The liposomal adjuvant formulations also increased the production of antigen-specific antibodies in the circulation following vaccine challenge. The results additionally show that CpG and poly(I:C) target distinct pathways in afferent lymph to induce their immunological effects, where CpG uniquely increased dendritic-cell associated antigen transport and induced the maturation of monocytes and dendritic cells 72h after injection. CpG also induced the persistence of gene programs involved in cell migration, intracellular DNA sensing and cytotoxic immunity in afferent lymph at this time point. These immunological effects were not observed with liposomal poly(I:C) or liposomes alone. This further translated into an extended period of lymph node cell shut down, the induction of IFNγ positive T cells in efferent lymph and enhanced production of antigen-specific antibodies after injection of liposomal CpG when compared to liposomal poly(I:C) and liposomes alone. The development of a preliminary mathematical…