Rational Design of DNA-Expressed Stabilized Native-Like HIV-1 Envelope Trimers

Aldon, Yoann; McKay, Paul F.; Allen, Joel D.; Ozorowski, Gabriel; Lévai, R.; Tolazzi, Monica; Rogers, P. S. Z.; He, Linling; Val, Natalia de; Fábián, Katalin; Scarlatti, Gabriella; Zhu, Jiang; Ward, Andrew B.; Crispin, Max; Shattock, Robin J.

doi:10.1016/j.celrep.2018.08.051

Cited by 53 publications

(108 citation statements)

References 68 publications

(118 reference statements)

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“…The occlusion of non-Nab epitopes may refocus the immune system toward NAb targets. This concept is well established in HIV vaccine development where mutations have been designed to stabilize the HIV Env trimer and occlude non-NAb epitopes with some success in eliciting NAb responses in small animals (28,29).…”

Section: Discussionmentioning

confidence: 99%

Enhancing the antigenicity and immunogenicity of monomeric forms of hepatitis C virus E2 for use as a preventive vaccine

Center

Boo

Phu

et al. 2020

Journal of Biological Chemistry

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The E2 glycoprotein of hepatitis C virus (HCV) is the major target of broadly neutralizing antibodies (bNAbs) that are critical for the efficacy of a prophylactic HCV vaccine. We previously showed that a cell culture–derived, disulfide-linked high-molecular-weight (HMW) form of the E2 receptor–binding domain lacking three variable regions, Δ123-HMW, elicits broad neutralizing activity against the seven major genotypes of HCV. A limitation to the use of this antigen is that it is produced only at low yields and does not have a homogeneous composition. Here, we employed a sequential reduction and oxidation strategy to efficiently refold two high-yielding monomeric E2 species, D123 and a disulfide-minimized version (D123A7), into disulfide-linked HMW-like species (Δ123r and Δ123A7r). These proteins exhibited normal reactivity to bNAbs with continuous epitopes on the neutralizing face of E2, but reduced reactivity to conformation-dependent bNAbs and nonneutralizing antibodies (non-NAbs) compared with the corresponding monomeric species. Δ123r and Δ123A7r recapitulated the immunogenic properties of cell culture–derived D123-HMW in guinea pigs. The refolded antigens elicited antibodies that neutralized homologous and heterologous HCV genotypes, blocked the interaction between E2 and its cellular receptor CD81, and targeted the AS412, AS434, and AR3 domains. Of note, antibodies directed to epitopes overlapping with those of non-NAbs were absent. The approach to E2 antigen engineering outlined here provides an avenue for the development of preventive HCV vaccine candidates that induce bNAbs at higher yield and lower cost.

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Section: Discussionmentioning

confidence: 99%

Enhancing the antigenicity and immunogenicity of monomeric forms of hepatitis C virus E2 for use as a preventive vaccine

Center

Boo

Phu

et al. 2020

Journal of Biological Chemistry

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“…Significant work has been put into stabilizing soluble HIV-1 Envelope (Env) immunogens to make them amenable to the clinic and large scale manufacturing (12)(13)(14)(15). We and others have developed stabilized native-like trimers that can be delivered through DNA or RNA platforms (15)(16)(17) and immune responses induced by these trimers could be polished by recombinant protein boost immunization(s) at sites of interest. Nucleic acid-based, and in particular DNA-based, vaccines offer a great opportunity by limiting the extent of manufacturing and long-term storage considerations.…”

mentioning

confidence: 99%

Chemokine-Adjuvanted Plasmid DNA Induces Homing of Antigen-Specific and Non–Antigen-Specific B and T Cells to the Intestinal and Genital Mucosae

Aldon

Shattock

McKay

2020

The Journal of Immunology

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Plasmid DNA is a promising vaccine platform that together with electroporation can elicit significant systemic Ab responses; however, immunity at mucosal sites remains low. In this study, we sought to program T and B cells to home to the gastrointestinal and vaginal mucosae using genetic chemokine adjuvants and assessed their impact on immune homeostasis in various distinct immune compartments. BALB/c mice were immunized i.m. with plasmid DNA encoding a model Ag HIV-1 Env gp140 and selected chemokines/cytokine and boosted intravaginally with gp140 recombinant protein. Isolated splenocytes, intestinal lymphocytes, and genital lymphocytes as well as serum and intestinal luminal contents were assessed for Ag-specific reactivity. In addition, flow cytometric analysis was performed to determine the impact on immune homeostasis at these sites. Different molecular chemokine/ cytokine adjuvants effected significant alterations to the recruitment of B and T cells to the spleen, vaginal and intestinal mucosae, for example CCL25 enhanced splenic and vaginal Ag-specific T cell responses whereas CCL28 increased the levels of specific T cells only in the vaginal mucosa. The levels of Ab could be modulated in the systemic circulation, as well as the vaginal vault and intestinal lumen, with CCL20 playing a central role. Our data demonstrate that the CCL20, CCL25, and CCL28 genetic chemokine adjuvants enhance the vaccine Ag-specific humoral and cellular responses and induce homing to the intestinal and female genital mucosae.

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“…Additionally, the SOSIP trimers did not bind at all to non-NAbs particularly in gp120 V3 and some CD4bs epitopes, as well as possessed slightly less complex and less extensively processed glycans compared to the same full-length trimer [46][47][48]. Although no UFO trimer has been characterized using this FRETbased dynamic approach, the Env bearing the UFO design has been proven to display similar antigenicity to its native-like, membrane-bound form [49]. Given the outstanding performance of the UFO design in trimer formation, it will be important to investigate the dynamics of UFO trimers, as well as SOSIP and NFL trimers, in comparison with Envs on matured virions.…”

Section: Discussionmentioning

confidence: 98%

Stabilized diverse HIV-1 envelope trimers for vaccine design

Wang

Liang

et al. 2020

Emerging Microbes & Infections

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One of the major goals in HIV-1 vaccine development is to achieve properly folded and stabilized envelope glycoprotein (Env) trimers that mimic the native Env on the mature virion. Here, we design and characterize uncleaved prefusionoptimized (UFO) trimers for 12 Envs currently circulating in China. Biochemical and biophysical characterization of these UFO trimers identified two subtype B/Bʹ Envs, CNE6 and MG13, which exhibited the highest trimer content and stability at a level comparable to the subtype A reference, BG505. Replacing the gp41 ectodomain (gp41 ECTO) of CRF01_AE trimers with that of CNE6, MG13, and BG505 resulted in chimeric constructs with significantly improved trimer content and stability. Negative-stain electron microscopy (EM) confirmed the structural integrity of these chimeric UFO trimers with CNE6 gp41 ECTO. Antibody binding assays showed that the chimeric trimers shared similar antigenic profiles to those with their original gp41 ECTO domains. Our results thus revealed the intrinsic differences among HIV-1 Envs of diverse origins and the critical role of gp41 ECTO in stabilizing the trimeric spike. By taking advantage of naturally stable Envs, gp41 ECTO swapping may represent a universal approach for the generation of stable trimers with the desired structural and antigenic properties for downstream in vivo evaluation and vaccine development.

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Rational Design of DNA-Expressed Stabilized Native-Like HIV-1 Envelope Trimers

Cited by 53 publications

References 68 publications

Enhancing the antigenicity and immunogenicity of monomeric forms of hepatitis C virus E2 for use as a preventive vaccine

Enhancing the antigenicity and immunogenicity of monomeric forms of hepatitis C virus E2 for use as a preventive vaccine

Chemokine-Adjuvanted Plasmid DNA Induces Homing of Antigen-Specific and Non–Antigen-Specific B and T Cells to the Intestinal and Genital Mucosae

Stabilized diverse HIV-1 envelope trimers for vaccine design

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