The Development of Contemporary Antigen Presenting Cell-Targeting Gene Delivery Vectors for the Generation of a New Class of Vaccines.

摘要

Genetic vaccines offer a treatment opportunity based upon successful gene delivery to specific immune cell modulators. Driving the process is the vector chosen for gene cargo packaging and subsequent delivery to antigen presenting cells (APCs) capable of triggering an immune cascade. As such, the delivery process must successfully navigate a series of requirements and obstacles associated with the chosen vector and target cell. However, most vectors currently utilized rely on ubiquitous delivery mechanisms that ineffectively target important immune effectors such as antigen presenting cells (APCs). Furthermore, APC targeting enables the ability to tune the direction (humoral vs cell-mediated) and strength of the resulting immune responses. Thus, in this dissertation, work is presented describing the development, refinement, and in vivo assessment of two distinct APC-targeting gene delivery strategies. Specifically, mannosylated poly(beta-amino esters) (PBAEs) and hybrid gene delivery vectors containing biological and biomaterial components. Each strategy demonstrated positive immunological outcomes in initial in vitro assessments using methodologies (developed in this dissertation) designed to provide a more a more realistic physiochemical profile prior to in vivo evaluations. Formulations of each strategy that displayed results significantly higher than all known commercial controls were chosen to continue to in vivo gene delivery vaccine models. Moreover, both strategies demonstrated strong, efficient, and safe in vivo humoral immune response profiles without the use of adjuvants when compared to genetic and protein control antigens. In summary, the gene delivery effectiveness provided by both mannosylated PBAE and Hybrid vectors offers specificity and potency in directing APC activation and subsequent Immune responses.