A Tale of Two Fusion Proteins: Understanding the Metastability of Human Respiratory Syncytial Virus and Metapneumovirus and Implications for Rational Design of Uncleaved Prefusion-closed Trimers

Overview

Journal bioRxiv

Date 2024 Mar 18

PMID 38496645

Authors

Yi-Zong Lee

Jerome Han

Yi-Nan Zhang

Garrett Ward

Keegan Braz Gomes

Sarah Auclair

Robyn L Stanfield

Linling He

Ian A Wilson

Jiang Zhu

Affiliations

Soon will be listed here.

Abstract

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) cause human respiratory diseases and are major targets for vaccine development. In this study, we designed uncleaved prefusion-closed (UFC) trimers for the fusion (F) proteins of both viruses by examining mutations critical to F metastability. For RSV, we assessed four previous prefusion F designs, including the first and second generations of DS-Cav1, SC-TM, and 847A. We then identified key mutations that can maintain prefusion F in a native-like, closed trimeric form (up to 76%) without introducing any interprotomer disulfide bond. For hMPV, we developed a stable UFC trimer with a truncated F-F linkage and an interprotomer disulfide bond. Tens of UFC constructs were characterized by negative-stain electron microscopy (nsEM), x-ray crystallography (11 RSV-F and one hMPV-F structures), and antigenic profiling. Using an optimized RSV-F UFC trimer as bait, we identified three potent RSV neutralizing antibodies (NAbs) from a phage-displayed human antibody library, with a public NAb lineage targeting sites Ø and V and two cross-pneumovirus NAbs recognizing site III. In mouse immunization, rationally designed RSV-F and hMPV-F UFC trimers induced robust antibody responses with high neutralizing titers. Our study provides a foundation for future prefusion F-based RSV and hMPV vaccine development.

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