When the MERS CoV outbreak raised global health concerns, we initiated a program to develop a candidate vaccine. Starting from Spike glycoprotein (S) sequences, we developed an immunization strategy consisting of a full-length S DNA prime and a S1 subunit protein boost that elicited high titers of neutralizing antibodies against eight different MERS-CoV strains. Immune sera contained potent neutralizing antibodies targeting the receptor binding domain (RBD), non-RBD portions of S1, and the S2 subunit. From the immunized mice, we produced a panel of hybridomas and produced monoclonal antibodies from which a variety with high neutralizing activity were selected for further characterization. The atomic structure of a monoclonal antibody, D12, in complex with the RBD revealed two distinct mechanisms by which they block binding to the MERS-CoV receptor, DPP4. In addition, immunogenicity was measured in nonhuman primates. Thus, vaccine immunogens designed from S sequences induced a diverse repertoire of neutralizing antibodies, demonstrating an efficient approach to vaccine design that may be applicable to other emerging viruses. Structural studies were also initiated with the spike glycoprotein of the HKU1 beta-coronavirus, to explore structure and for receptor discovery and led to a spike trimer structure solution. Human airway epithelial cell culture system was established to initiate entry and pathogenesis studies of HKU1. Through collaborations, we have solved the structure of the MERS S protein and based on this structure, designed stabilizing mutations which stabilize the S protein in its prefusion conformation. We are currently assessing immunogenicity of the stabilized MERS S protein, and evaluating stabilizing mutations in other coronavirus spike proteins.
| Wang, Lingshu; Shi, Wei; Chappell, James D et al. (2018) Importance of neutralizing monoclonal antibodies targeting multiple antigenic sites on MERS-CoV Spike to avoid neutralization escape. J Virol : |
| Pallesen, Jesper; Wang, Nianshuang; Corbett, Kizzmekia S et al. (2017) Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen. Proc Natl Acad Sci U S A 114:E7348-E7357 |
| Kirchdoerfer, Robert N; Cottrell, Christopher A; Wang, Nianshuang et al. (2016) Pre-fusion structure of a human coronavirus spike protein. Nature 531:118-21 |
| Wang, Lingshu; Shi, Wei; Joyce, M Gordon et al. (2015) Evaluation of candidate vaccine approaches for MERS-CoV. Nat Commun 6:7712 |
