TY - JOUR
T1 - Cleavage-Independent HIV-1 Env Trimers Engineered as Soluble Native Spike Mimetics for Vaccine Design
AU - Sharma, Shailendra K.
AU - deVal, Natalia
AU - Bale, Shridhar
AU - Guenaga, Javier
AU - Tran, Karen
AU - Feng, Yu
AU - Dubrovskaya, Viktoriya
AU - Ward, Andrew B.
AU - Wyatt, Richard T.
N1 - Funding Information:
We thank John Mascola, Marshall Posner, James Robinson, and Dennis Burton for providing monoclonal antibodies, Pascal Poignard and Alejandra Ramos for providing BG505 gp120, and Richard Wilson for providing JRFL gp140-foldon. This study was supported by funding from the NIH HIVRAD (AI104722), the Scripps CHAVI-ID (AI100663), and IAVI, which is made possible by the support of many donors, including the Bill & Melinda Gates Foundation, the Ministry of Foreign Affairs of Denmark, Irish Aid, the Ministry of Finance of Japan, the Ministry of Foreign Affairs of the Netherlands, the Norwegian Agency for Development Cooperation (NORAD), the United Kingdom Department for International Development (DFID), and the United States Agency for International Development (USAID). The full list of IAVI donors is available at http://www.iavi.org . This study is made possible by the generous support of the Bill & Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery and the American people through USAID. The contents of this manuscript are the responsibility of IAVI and do not necessarily reflect the views of USAID or the United States government.
Publisher Copyright:
© 2015 The Authors.
PY - 2015/4/28
Y1 - 2015/4/28
N2 - Viral glycoproteins mediate entry by pH-activated or receptor-engaged activation and exist in metastable pre-fusogenic states that may be stabilized by directed rational design. As recently reported, the conformationally fixed HIV-1 envelope glycoprotein (Env) trimers in the pre-fusion state (SOSIP) display molecular homogeneity and structural integrity at relatively high levels of resolution. However, the SOSIPs necessitate full Env precursor cleavage, which requires endogenous furin overexpression. Here, we developed an alternative strategy using flexible peptide covalent linkage of Env subdomains to produce soluble, homogeneous, and cleavage-independent Env mimics, called native flexibly linked (NFL) trimers, as vaccine candidates. This simplified design avoids the need for furin co-expression and, in one case, antibody affinity purification to accelerate trimer scale-up for preclinical and clinical applications. We have successfully translated the NFL design to multiple HIV-1 subtypes, establishing the potential to become a general method of producing native-like, well-ordered Env trimers for HIV-1 or other viruses.
AB - Viral glycoproteins mediate entry by pH-activated or receptor-engaged activation and exist in metastable pre-fusogenic states that may be stabilized by directed rational design. As recently reported, the conformationally fixed HIV-1 envelope glycoprotein (Env) trimers in the pre-fusion state (SOSIP) display molecular homogeneity and structural integrity at relatively high levels of resolution. However, the SOSIPs necessitate full Env precursor cleavage, which requires endogenous furin overexpression. Here, we developed an alternative strategy using flexible peptide covalent linkage of Env subdomains to produce soluble, homogeneous, and cleavage-independent Env mimics, called native flexibly linked (NFL) trimers, as vaccine candidates. This simplified design avoids the need for furin co-expression and, in one case, antibody affinity purification to accelerate trimer scale-up for preclinical and clinical applications. We have successfully translated the NFL design to multiple HIV-1 subtypes, establishing the potential to become a general method of producing native-like, well-ordered Env trimers for HIV-1 or other viruses.
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U2 - 10.1016/j.celrep.2015.03.047
DO - 10.1016/j.celrep.2015.03.047
M3 - Article
C2 - 25892233
AN - SCOPUS:84928583366
VL - 11
SP - 539
EP - 550
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 4
ER -