Domain III of the envelope protein as a dengue vaccine target

Guzmán, María G.; Hermida, Lisset; Bernardo, Lídice; Ramírez, Rosa; Guillén, Gerardo

doi:10.1586/erv.09.139

Cited by 112 publications

(73 citation statements)

References 96 publications

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“…These results demonstrate that 2 or 3 immunizations with the TDV elicited robust, high-titer, neutralizing antibody responses, particularly for DENV-1 and DENV-2, with somewhat lower titers for DENV-3 and DENV-4. One of the major concerns for the development of EIII and other subunit dengue vaccines is the lack of durable antibody responses (18,31). We determined the longevity of the neutralizing antibody responses in mice for a TDV candidate consisting of R3.2xD1EIII, R3.2xD2EIII, R3.2xD3EIII, and R3.2xD4EIII.…”

Section: Resultsmentioning

confidence: 99%

“…The carboxyl-terminal E domain III (EIII) is the smallest domain (approximately 100 amino acids in length) and contains the putative receptor binding site as well as type-specific and subcomplex-specific neutralizing epitopes that are recognized by strongly neutralizing antibodies (11,(14)(15)(16)(17), making it an attractive target for subunit vaccine development. However, EIII fusion proteins delivered by plasmid DNA or purified from Escherichia coli or baculovirus expression systems are often poorly immunogenic, requiring relatively high antigen doses or potentially toxic adjuvants to achieve moderate immunogenicity and protective efficacy in animal models (18).VaxInnate has developed a clinically proven recombinant flagellin-antigen fusion platform to allow rapid development and economical manufacturing of fusion protein-based vaccines using a well-established E. coli fermentation system and a standardized purification process. This vaccine platform contains bacterial …”

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confidence: 99%

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Immunogenicity and Efficacy of Flagellin-Envelope Fusion Dengue Vaccines in Mice and Monkeys

Liu

Song

Putnak

et al. 2015

Clin. Vaccine Immunol.

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The envelope (E) protein of flaviviruses includes three domains, EI, EII, and EIII, and is the major protective antigen. Because EIII is rich in type-specific and subcomplex-specific neutralizing epitopes and is easy to express, it is particularly attractive as a recombinant vaccine antigen. VaxInnate has developed a vaccine platform that genetically links vaccine antigens to bacterial flagellin, a Toll-like receptor 5 ligand. Here we report that tetravalent dengue vaccines (TDVs) consisting of four constructs, each containing two copies of EIII fused to flagellin (R3.2x format), elicited robust and long-lived neutralizing antibodies (geometric mean titers of 200 to 3,000), as measured with a 50% focus reduction neutralization test (FRNT 50 ). In an immunogenicity study, rhesus macaques (n ‫؍‬ 2) immunized subcutaneously with 10 g or 90 g of TDV three or four times, at 4-to 6-week intervals, developed neutralizing antibodies to four dengue virus (DENV) serotypes (mean post-dose 3 FRNT 50 titers of 102 to 601). In an efficacy study, rhesus macaques (n ‫؍‬ 4) were immunized intramuscularly with 16 g or 48 g of TDV or a placebo control three times, at 1-month intervals. The animals that received 48-g doses of TDV developed neutralizing antibodies against the four serotypes (geometric mean titers of 49 to 258) and exhibited reduced viremia after DENV-2 challenge, with a group mean viremia duration of 1.25 days and 2 of 4 animals being completely protected, compared to the placebo-treated animals, which all developed viremia, with a mean duration of 4 days. In conclusion, flagellin-EIII fusion vaccines are immunogenic and partially protective in a nonhuman primate model. Dengue disease poses a significant health threat to almost onehalf of the world's population residing in the Asian-Pacific region and Latin America, where the disease is endemic, as well as to travelers and military personnel. Currently, no licensed dengue vaccine is available. Several vaccine candidates based on live attenuated dengue viruses (DENVs) or dengue virus-flavivirus chimeras are currently being evaluated in phase II or III clinical trials (for reviews, see references 1-3). However, some of these vaccines require lengthy immunization regimens of up to 1 year for the development of immunity to all four serotypes, which seems to make them less well suited for travelers and military personnel. In a series of recently published phase IIb/III trials conducted in five Asian countries in which dengue is endemic, the Sanofi-Aventis chimeric yellow fever (YF) 17D-DENV-1 to -4 CYD tetravalent dengue vaccine (TDV) showed an acceptable safety profile and 56% overall efficacy (4); however, the vaccine failed to confer significant protection against DENV-2 (5). These findings underscore the need for the development of alternative vaccine platforms, such as those based on purified inactivated virus (6), DNA (7), and recombinant subunit truncated envelope (E) proteins (e.g., 80% envelope protein [80E]) (8). Some of these alternative vaccine candidates hav...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Immunogenicity and Efficacy of Flagellin-Envelope Fusion Dengue Vaccines in Mice and Monkeys

Liu

Song

Putnak

et al. 2015

Clin. Vaccine Immunol.

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“…In contrast, the LR mutant viruses were no longer neutralized by the EDIII vaccine serum. Further studies are needed to understand why immunization with the ectodomain of E protein stimulated EDIII lateral ridge but not A strand binding neutralizing antibodies, especially because some groups are pursuing recombinant E protein antigens as dengue vaccine candidates (2,11,14). We conclude that while EDIII-targeted immunizations can stimulate an LR-focused neutralizing antibody response, most neutralizing antibodies in people exposed to DENV2 infections are not directed against these epitopes.…”

mentioning

confidence: 82%

Recombinant Dengue Type 2 Viruses with Altered E Protein Domain III Epitopes Are Efficiently Neutralized by Human Immune Sera

Wahala

Huang

Butrapet

et al. 2012

J Virol

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cHumans develop polyclonal, serotype-specific neutralizing antibody responses after dengue virus (DENV) infection. Many mouse antibodies that neutralize DENV bind to the lateral ridge or A strand epitopes on domain III of the viral envelope (EDIII) protein. It has been assumed that these epitopes are also the main target of human neutralizing antibodies. Using recombinant dengue serotype 2 viruses with altered EDIII epitopes, we demonstrate that EDIII epitopes are not the main target of human neutralizing antibody. Dengue viruses (DENV) are emerging, mosquito-borne flaviviruses of humans (10). Some people infected with dengue have asymptomatic or mild disease, while others develop classical dengue fever (DF) or dengue hemorrhagic fever (DHF), which can be fatal (12, 13). The DENV complex consists of 4 distinct but related viruses designated as serotypes. Although infection with one serotype stimulates an adaptive immune response that is highly cross-reactive between serotypes, this response only prevents reinfection with the homologous serotype (13). People experiencing a second infection with a new serotype face a much greater risk of developing DHF because preexisting, cross-reactive immunity can exacerbate disease. A leading theory to explain the association between preexisting immunity and severe disease is antibody-dependent enhancement (ADE), which postulates that cross-reactive, weakly neutralizing antibodies enhance the ability of DENVs to infect Fc receptor-bearing cells (13) and the amount of virions released from each infected cell (32). Antibodies also play a key role in neutralizing DENVs and appear to provide longterm protection from reinfection. Currently, several live attenuated dengue vaccines are being tested in clinical trials. Despite the advanced stage of live DENV vaccine development, we do not know the properties of human antibodies responsible for potent and long-term neutralization following natural infection.The DENV envelope (E) protein that covers the surface of the virion is the main target of neutralizing antibodies. Each folded E protein molecule contains three distinct domains, designated EDI, EDII, and EDIII (Fig. 1A) (21-23). Most mouse monoclonal antibodies (MAbs) that strongly neutralize DENVs bind to epitopes on the lateral ridge (LR) and A strand of EDIII (Fig. 1B) (26,28,30,31). The LR epitope is poorly conserved between DENV serotypes, and antibodies that target this epitope only bind and neutralize a single serotype (dengue type specific) (8, 31). The A strand epitope is more conserved between serotypes, and antibodies that bind to this epitope usually bind and neutralize more than one serotype (dengue subcomplex) (31). Some antibodies are sensitive to mutations in both the LR and A strand, indicating that the footprints of these antibodies span both epitopes (8,9,19,31).As people exposed to DENV infections develop strong, typespecific or subcomplex-neutralizing polyclonal antibody responses, it was plausible that human neutralizing antibodies would also bind to epitopes ...

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“…The envelop protein domain III (EDIII) induces serotype-specific antibodies. Although it has low intrinsic potential for eliciting cross-reactive antibodies against heterologous serotypes (Hombach et al 2005), it has emerged as the most promising region for subunit vaccine development (Guzman et al 2010). Recombinant antigens based on EDIII have been produced using bacteria, yeast, insect cells and plants (Batra et al 2010b;Cardoso et al 2013;Clements et al 2010;Etemad et al 2008;Martinez et al 2010;Saejung et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Production of dengue virus envelope protein domain III-based antigens in tobacco chloroplasts using inducible and constitutive expression systems

et al. 2016

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Dengue fever is a disease in many parts of the tropics and subtropics and about half the world's population is at risk of infection according to the World Health Organization. Dengue is caused by any of the four related dengue virus serotypes DEN-1, -2, -3 and -4, which are transmitted to people by Aedes aegypti mosquitoes. Currently there is only one vaccine (Dengvaxia Ò ) available (limited to a few countries) on the market since 2015 after half a century's intensive efforts. Affordable and accessible vaccines against dengue are hence still urgently needed. The dengue envelop protein domain III (EDIII), which is capable of eliciting serotype-specific neutralizing antibodies, has become the focus for subunit vaccine development. To contribute to the development of an accessible and affordable dengue vaccine, in the current study we have used plant-based vaccine production systems to generate a dengue subunit vaccine candidate in tobacco. Chloroplast genome engineering was applied to express serotypespecific recombinant EDIII proteins in tobacco chloroplasts using both constitutive and ethanol-inducible expression systems. Expression of a tetravalent antigen fusion construct combining EDIII polypeptides from all four serotypes was also attempted. Transplastomic EDIIIexpressing tobacco lines were obtained and homoplasmy was verified by Southern blot analysis. Northern blot analyses showed expression of EDIII antigen-encoding genes. EDIII protein accumulation levels varied for the different recombinant EDIII proteins and the different expression systems, and reached between 0.8 and 1.6 % of total cellular protein. Our study demonstrates the suitability of the chloroplast compartment as a production site for an EDIII-based vaccine candidate against dengue fever and presents a Gateway Ò plastid transformation vector for inducible transgene expression.

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Domain III of the envelope protein as a dengue vaccine target

Cited by 112 publications

References 96 publications

Immunogenicity and Efficacy of Flagellin-Envelope Fusion Dengue Vaccines in Mice and Monkeys

Immunogenicity and Efficacy of Flagellin-Envelope Fusion Dengue Vaccines in Mice and Monkeys

Recombinant Dengue Type 2 Viruses with Altered E Protein Domain III Epitopes Are Efficiently Neutralized by Human Immune Sera

Production of dengue virus envelope protein domain III-based antigens in tobacco chloroplasts using inducible and constitutive expression systems

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