Thermostability of Well-Ordered HIV Spikes Correlates with the Elicitation of Autologous Tier 2 Neutralizing Antibodies

PLoS Pathogens

Karen Tran¹  Shailendra Kumar¹  Joanne DeStefano²  Andrew B. Ward³  Richard T. Wyatt³  Heather Arendt⁴  Natalia de Val⁴  Yu Feng⁴  Shridhar Bale⁴  Richard Wilson⁴  Javier Guenaga⁵ 

[1]Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, United States of America

[2]Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, United States of America

[3]IAVI Design and Development Lab, Department of Research and Development, International AIDS Vaccine Initiative, Brooklyn, New York, United States of America

[4]IAVI Neutralizing Center at TSRI, Department of Research and Development, International AIDS Vaccine Initiative, La Jolla, California, United States of America

[5]The Scripps CHAVI-ID, The Scripps Research Institute, La Jolla, California, United States of America

关键词: Antibodies;    Enzyme-linked immunoassays;    Antigens;    Cross-linking;    Immunologic adjuvants;    Blue native polyacrylamide gel electrophoresis;    Guinea pigs;    B cells;   

DOI  :  10.1371/journal.ppat.1005767

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

In the context of HIV vaccine design and development, HIV-1 spike mimetics displaying a range of stabilities were evaluated to determine whether more stable, well-ordered trimers would more efficiently elicit neutralizing antibodies. To begin, in vitro analysis of trimers derived from the cysteine-stabilized SOSIP platform or the uncleaved, covalently linked NFL platform were evaluated. These native-like trimers, derived from HIV subtypes A, B, and C, displayed a range of thermostabilities, and were “stress-tested” at varying temperatures as a prelude to in vivo immunogenicity. Analysis was performed both in the absence and in the presence of two different adjuvants. Since partial trimer degradation was detected at 37°C before or after formulation with adjuvant, we sought to remedy such an undesirable outcome. Cross-linking (fixing) of the well-ordered trimers with glutaraldehyde increased overall thermostability, maintenance of well-ordered trimer integrity without or with adjuvant, and increased resistance to solid phase-associated trimer unfolding. Immunization of unfixed and fixed well-ordered trimers into animals revealed that the elicited tier 2 autologous neutralizing activity correlated with overall trimer thermostability, or melting temperature (Tm). Glutaraldehyde fixation also led to higher tier 2 autologous neutralization titers. These results link retention of trimer quaternary packing with elicitation of tier 2 autologous neutralizing activity, providing important insights for HIV-1 vaccine design.

【 授权许可】

CC BY