Veterinary and Biomedical Sciences, Department of
Date of this Version
5-2016
Abstract
The sole envelope glycoprotein spike on the surface of Human Immunodeficiency Virus (HIV-1) is composed of two subunits: gp120 and gp41. The gp120 consists of three domains: the inner domain, outer domain and bridging sheet, to which the viral primary receptor CD4 binds and induces a conformational change in gp120 to expose the co-receptor binding site. Previous studies have found that the outer domain of gp120 is relatively more stable than the inner domain and bridging sheet. When superimposed, the co-crystal structures of CD4-gp120 complex and VRC01-gp120 complex revealed that the CD4-binding site (CD4-BS) antibody VRC01 actually mainly bound to the outer domain of the gp120. The outer domain (designed OD1) of gp120 has been considered as an immunogen candidate but to be ineffective, suggesting that structural-based modifications to the outer domain are required for designing an effective vaccine immunogen. Previous studies in our laboratory have introduced two disulfide bonds to stabilize the outer domain structure and named it OD2, which significantly improved the immunogenicity when compared to the wild-type outer domain (OD1). In this project, based on the OD2 set of experiment, an additional important mutation from serine to tryptophan at position 375 (S375W) in the CD4-binding cavity was introduced to further stabilize the outer domain, which was named OD3. The OD3 immunogen was expressed and purified in E. coli. Subsequently, the purified OD3 immunogen was characterized in vitro and its immunogenicity was tested in guinea pigs. In comparison to OD1 and OD2, the results demonstrate the antibody titers and neutralizing activity were clearly higher in OD3. It is suggested that the S375W mutation stabilized the outer domain and further improved the outer domain (OD) immunogenicity. In conclusion, this research has used a structure-based approach to design HIV-1-1 vaccine immunogens based on the gp120 outer domain (OD).
Advisor: Shi-hua Xiang
Comments
A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science, Major: Veterinary Science, Under the Supervision of Professor Shi-hua Xiang. Lincoln, Nebraska: May, 2016
Copyright (c) 2016 Duoyi Hu