Human antibody response to Zika targets type-specific quaternary structure epitopes

Collins, Matthew H.; Tu, Huy A.; Gimblet-Ochieng, Ciara; Liou, Guei-Jiun Alice; Jadi, Ramesh; Metz, Stefan; Thomas, Ashlie; McElvany, Benjamin D.; Davidson, Edgar; Doranz, Benjamin J.; Reyes, Yaoska; Bowman, Natalie M.; Becker‐Dreps, Sylvia; Bucardo, Filemón; Lazear, Helen M.; Diehl, Sean A.; Silva, Aravinda M. de

doi:10.1172/jci.insight.124588

Cited by 50 publications

(77 citation statements)

References 83 publications

(113 reference statements)

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“…We conclude that the rE M antigen induces an EDIII focused antibody response that is poorly neutralizing, whereas rE D antigens stimulate robust neutralizing antibody responses that target epitopes distinct from simple EDIII epitopes. We recently isolated two ZIKV-specific strongly neutralizing human mAbs (A9E and G9E) from a person who was infected with ZIKV in 2016 23 . These antibodies map to complex epitopes on domain I (A9E) and domain II (G9E) of ZIKV E protein 23 .…”

Section: Resultsmentioning

confidence: 99%

“…We recently isolated two ZIKV-specific strongly neutralizing human mAbs (A9E and G9E) from a person who was infected with ZIKV in 2016 23 . These antibodies map to complex epitopes on domain I (A9E) and domain II (G9E) of ZIKV E protein 23 . Moreover, antibody blockade assays using these mAbs and human immune sera indicated that the epitopes targeted by these mAbs also are targeted by antibodies in immune serum from Zika patients but not dengue patients 23 .…”

Section: Resultsmentioning

confidence: 99%

“…These antibodies map to complex epitopes on domain I (A9E) and domain II (G9E) of ZIKV E protein 23 . Moreover, antibody blockade assays using these mAbs and human immune sera indicated that the epitopes targeted by these mAbs also are targeted by antibodies in immune serum from Zika patients but not dengue patients 23 . We used a blockade of binding (BOB) assay to test if mice immunized with rE M or rE D developed antibodies that recognize A9E or G9E epitopes on ZIKV.…”

Section: Resultsmentioning

confidence: 99%

“…Mice were monitored for weight loss and ZIKV induced disease, and mice were bled at day 0 (before virus challenge), day 4 and day 14 ( Fig.4A ). The potently neutralizing mAb G9E was used as a positive control for protection against ZIKV infection 23 . Mice that received G9E or serum from rE D +alum immunization were completely protected against weight loss and disease signs.…”

Section: Resultsmentioning

confidence: 99%

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Oligomeric state of the ZIKV E protein defines protective immune responses

Metz

Thomas

Brackbill

et al. 2019

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8The current leading Zika vaccine candidates in clinical testing are based on live or killed virus 9 platforms, which have safety issues, especially in pregnant women. Zika subunit vaccines, 10 however, have shown poor performance in preclinical studies. We hypothesized that Zika 11Envelope (E) protein subunit vaccines have performed poorly because the antigens tested have 12been recombinant E monomers that do not display critical quaternary structure epitopes present 13on Zika E protein homodimers that cover the surface of the virus. To test this hypothesis, we 14 engineered and produced stable recombinant E protein homodimers. Unlike the E monomer, the 15 dimer was recognized by strongly neutralizing monoclonal antibodies isolated from Zika-immune 16 individuals. In a mouse model of vaccination, the dimeric antigen stimulated strongly neutralizing 17 antibodies that targeted epitopes that were similar to epitopes recognized by human antibodies 18 following natural Zika virus infection. In contrast, the monomer antigen stimulated lower levels of 19 neutralizing antibodies directed to simple epitopes on domain III of E protein. In a mouse model 20 of ZIKV challenge, only E dimer antigen stimulated protective antibodies, not the 21 monomer. These results highlight the importance of mimicking the highly structured flavivirus 22 surface when designing subunit vaccines. The flavivirus field has a long history of using E 23 monomers as vaccine antigens with limited success. These results are applicable to developing 24 second generation subunit vaccines against Zika as well as other medically important flaviviruses 25 such as dengue and yellow fever viruses. 26 130 Figure 3: ZIKV rE D stimulates antibodies that target complex epitopes on the virion. A) Method for 131 depleting EDIII binding antibodies from mouse immune sera. Recombinant ZIKV EDIII (His-tagged) was 132 coupled to nickel beads i then incubated with immune sera ii . Magnetic pull down removes nickel beads and 133 EDIII binding antibodies iii , leaving leavind sera depleted from EDIII binding antibodies iv . B) The level of 134 EDIII-binding antibodies in the serum of mice immunized with the indicated antigens is shown as a 135 percentage of the total level of ZIKV specific IgG, as measured by ELISA of EDIII-depleted and control-136 depleted serum. C) The neutralizing activity of EDIII-depleted and undepleted sera was determined and 137 expressed as the dilution at which 50% of the virus was neutralized (Neut50). D) A blockade of binding 138 (BOB) assay was used to evaluate if mice immunized with rE M or rE D developed antibodies that blocked 139 the binding of A9E, G9E and EDE C10 human mAbs. Data points represent individual mice. Statistical140differences were determined by one-way ANOVA followed by a Tukey's test (p<0.05).

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Oligomeric state of the ZIKV E protein defines protective immune responses

Metz

Thomas

Brackbill

et al. 2019

Preprint

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“…To further test the native antigenicity of ZIKV RVPs, we performed an ELISA with RVPs using various conformational MAbs. As detection MAbs, we used two EDE MAbs (C8 and C10) and five neutralizing anti-ZIKV MAbs (ZIKV-117, ZIKV-195, ZIKV-116 (29), A9E (28), and LM-081). Each of these seven MAbs represents a distinct conformational region on the native structure of ZIKV prM/E ( Fig.…”

Section: Zikv Rvps Are Infectious and Antigenically Identical To Livementioning

confidence: 99%

Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications

Whitbeck

Thomas

Kadash-Edmondson

et al. 2020

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The development of vaccines against flaviviruses, including Zika virus (ZIKV) and dengue virus (DENV), continues to be a major challenge, hindered by the lack of efficient and reliable methods for screening neutralizing activity of sera or antibodies. To address this need, we previously developed a plasmid-based, replication-incompetent DENV reporter virus particle (RVP) production system as an efficient and safe alternative to the Plaque Reduction Neutralization Test (PRNT). As part of the response to the 2015-2016 ZIKV outbreak, we recently developed pseudoinfectious ZIKV RVPs by modifying our DENV RVP system. The use of ZIKV RVPs as critical reagents in human clinical trials requires their further validation using stability and reproducibility metrics for large-scale applications. In the current study, we validated ZIKV RVPs using infectivity, neutralization, and enhancement assays with monoclonal antibodies (MAbs) and human ZIKV-positive patient serum. ZIKV RVPs are antigenically identical to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of plaque formation assays. We demonstrate reproducible neutralization titer data (NT50 values) across different RVP production lots, volumes, time frames, and laboratories. We also show RVP stability across experimentally relevant time intervals and temperatures. Our results demonstrate that ZIKV RVPs provide a safe, rapid, and reproducible reagent for large-scale, long-term studies of neutralizing antibodies and sera, which can facilitate large-scale screening and epidemiological studies to help expedite ZIKV vaccine development.

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