Sugar, Ice, and Everything (Not So) Nice: Structural Interrogation of the Lassa Glycoprotein Complex and Its Recognition by the Humoral Immune System

• Main Author: Perrett, Hailee R
• Format: Dissertation
• Language: English
• Published: ProQuest Dissertations & Theses 01-01-2023
• Subjects: Biophysics; Immunology; Virology
• ISBN: 9798380307413

Recognized as a priority emerging pathogen by the World Health Organization due to its potential to cause a public health crisis and the absence of efficacious therapeutics or vaccines, Lassa fever affects an estimated 300,000 people per year and results in approximately 5000 deaths. Lassa virus—a member of the Arenaviridae family—is decorated by heavily glycosylated envelope glycoprotein complexes (GPC) responsible for host recognition and viral fusion and as such are the focus of many preclinical vaccine studies. The trimeric conformation of GPC is necessary to induce almost all neutralizing antibody responses; however, the solubilized ectodomain is inherently unstable and has proved recalcitrant to unbound structural studies in the absence of stabilizing antibodies. To alleviate complexities associated with GPC’s metastable nature, we utilized a trimerization scaffold. This scaffold, which covalently attached via a flexible linker near the membrane proximal region of GPC, stabilizes the unliganded protein in its trimeric oligomerization state. We use this stabilization mechanism to advance our knowledge of the GPC, predominantly through a structural lens.The developments from this thesis can be summarized in two major categories: (1) the use of the stabilization scheme to develop a broader understanding of GPCs across Lassa lineages and (2) defining the interactions of antibodies with the GPC. In doing so, this work contributes to the foundation for further structure-guided therapeutic and immunogen design to combat Lassa fever.