Evaluation of recombinant dengue viral envelope B domain protein antigens for the detection of dengue complex-specific antibodies.

Simmons, Monika; Porter, Kevin R.; Escamilla, J. E.; Graham, Rachel L.; Watts, Douglas M.; Eckels, Kenneth H.; Hayes, Curtis G.

doi:10.4269/ajtmh.1998.58.144

Cited by 31 publications

(26 citation statements)

References 17 publications

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“…The presence of many type-specific epitopes in DIII and its relatively stable conformation suggest that it might be useful as a virus-specific antigen. It has been shown by others that DIII can be independently expressed and retain much of its biological activity (3,10,21,22,24). The results presented here correlate closely with previously published information concerning the antigenic structure of the flavivirus E protein; however, some alternative explanations for these results could be considered.…”

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confidence: 80%

Contribution of Disulfide Bridging to Epitope Expression of the Dengue Type 2 Virus Envelope Glycoprotein

Roehrig

Volpe

Squires

et al. 2004

J Virol

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The flavivirus envelope (E) glycoprotein encodes important phenotypic and immunogenic properties of the virion (7,17). This protein elicits virus-neutralizing antibodies, mediates virus-cell membrane fusion, and initiates infection through binding to cell surfaces. The high-resolution crystal structures of the ectodomain of the tick-borne encephalitis (TBE) virus and dengue type 2 (DEN2) virus E-protein homodimers have been solved (13, 16). The three structural domains (DI, DII, and DIII) identified in the E protein were roughly analogous to the previously defined antigenic domains (C, A, and B) (Fig. 1). The amino acid sequence of the ectodomain of all flavivirus E proteins contains 12 strictly conserved cysteine residues that form six disulfide (SS) bonds (14). For DEN2 virus, the SS1 bond (Cys 3 bonded to Cys 30) stabilizes an amino-terminal loop (Fig. 1). The SS2 (Cys 60-Cys 121), SS3 (Cys 74-Cys 105), and SS4 (Cys 92-Cys 116) bonds stabilize the amino-proximal part of DII that harbors the virus membrane fusion sequence located at amino acids (aa) 98 to 110 (17, 19). SS5 (Cys 185-Cys 285) stabilizes the carboxy-proximal loop of DII, which is separated from the amino-proximal DII loop by DI. The SS6 bond makes the only bridge in DIII (Cys 302-Cys 333).Reduction of all six of the West Nile virus E-protein SS bonds resulted in a protein that was unable to elicit virusneutralizing antibody in mice (23). To date, only DIII SS6 has been investigated individually for its contribution to the structure and function of the E protein. SS6 is of particular interest because of experimental evidence implicating DIII in cell attachment (5). By using proteolytic fragments of the TBE virus E protein, it was shown that SS6 spontaneously reformed following reduction and oxidation, suggesting that the surrounding secondary structure favored SS6 bond formation (25). This bond was also critical for the correct antigenic structure of the E proteins of DEN1 and DEN2 viruses (9, 12).The antigenic structure of the DEN2 virus E protein has been investigated previously using monoclonal antibodies (MAbs) (8,17). Not unexpectedly, the DEN2 virus E-protein antigenic structure was similar to the TBE virus E-protein antigenic structure. Three antigenic regions (A, B, and C) with unique biochemical characteristics were identified, and these regions correlated well with their analogous regions in the TBE virus E protein. The locations of many of the epitopes were mapped on the DEN2 E-protein three-dimensional structure using MAb competition binding assays, binding of MAb to peptide fragments, and serological testing of MAbs (17).In this study we used site-directed mutagenesis of a chimeric plasmid expressing the prM and E proteins of DEN2 virus strain 16681 to determine the individual contributions of the six SS bonds to E-protein epitope expression. This chimeric plasmid, pCB8D2-2J-2-9-1, expressed modified DEN2 proteins (4). The DEN2 prM signal sequence was replaced by the Japanese encephalitis virus prM signal sequence. The last 20% (aa 39...

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supporting

confidence: 80%

Contribution of Disulfide Bridging to Epitope Expression of the Dengue Type 2 Virus Envelope Glycoprotein

Roehrig

Volpe

Squires

et al. 2004

J Virol

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“…Earlier investigations with other flaviviruses have also reported the presence of virusspecific epitopes within this region of the E protein (e.g., see references 6, 22, and 25), and other authors have suggested the utility of domain III from dengue virus types 1 to 4 or JEV as antigens for specific serological diagnosis of infections with those flaviviruses (10,24). These observations led us to investigate the utility of a recombinant, bacterially expressed domain III (r-EIII) antigen derived from the envelope protein of a North American WNV strain (385-99) for discrimination of WNV from other JE virus group infections.…”

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confidence: 99%

Use of a Recombinant Envelope Protein Subunit Antigen for Specific Serological Diagnosis of West Nile Virus Infection

et al. 2004

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Serological diagnosis of West Nile virus (WNV) infection is complicated by extensive antigenic crossreactivity with other closely related flaviviruses, such as St. Louis encephalitis virus.Here we describe a recombinant, bacterially expressed antigen equivalent to structural domain III of the WNV envelope protein that has allowed clear discrimination of antibody responses to WNV from those against other related flaviviruses in indirect enzyme-linked immunosorbent assays using standardized control antisera and field-collected samples.Since 1999, the United States has experienced annual epidemics of disease in humans and animals caused by West Nile virus (WNV) over an expanding geographical range. To date during 2003, WNV has been isolated in 46 states and the District of Columbia, and more than 8,500 cases of human disease, resulting in 199 deaths, have been reported (1). Outbreaks of WNV disease with neurological manifestations have also been reported in Eastern Europe, North Africa, and Israel since the mid-1990s (reviewed in reference 17). WNV is clearly an emerging and significant public health problem. Research priorities to limit the impact of WNV include the development of more-specific rapid diagnostic assays (5, 18).WNV is a member of the Japanese encephalitis (JE) virus group of the genus Flavivirus, family Flaviviridae; other members include Japanese encephalitis virus (JEV), found throughout Asia, St. Louis encephalitis virus (SLEV), found in the Americas, and Murray Valley encephalitis virus (MVEV), found in Australia and New Guinea. These viruses have a similar ecology and are antigenically related to WNV, and their cocirculation in several regions of the world has complicated the specific diagnosis of infections by these viruses in humans and other vertebrate hosts (10, 15). Cross-reactions in patients ultimately diagnosed with probable dengue virus infections have also been reported in evaluations of WNV testing assays (19).Previously we have reported the identification of WNVspecific neutralizing epitopes within structural domain III of the WNV envelope (E) protein (2). Earlier investigations with other flaviviruses have also reported the presence of virusspecific epitopes within this region of the E protein (e.g., see references 6, 22, and 25), and other authors have suggested the utility of domain III from dengue virus types 1 to 4 or JEV as antigens for specific serological diagnosis of infections with those flaviviruses (10, 24). These observations led us to investigate the utility of a recombinant, bacterially expressed domain III (r-EIII) antigen derived from the envelope protein of a North American WNV strain (385-99) for discrimination of WNV from other JE virus group infections. MATERIALS AND METHODSExpression and purification of recombinant WNV E protein domain III. Regions corresponding to structural domain III of the WNV strain 385-99 E protein were reverse transcription-PCR amplified for cloning and expression either as a glutathione S-transferase (GST) fusion using the pGEX-2T syste...

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“…On glycoprotein E (10,14), a highly dengue virus typespecific antigenic side, the B domain or domain III, has been characterized (17). The short sequence of about 100 amino acids was expressed in Escherichia coli for the Japanese encephalitis (JE) virus by Mason et al (12) and for dengue viruses by Fonseca et al (4).…”

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Serological Differentiation of Infections with Dengue Virus Serotypes 1 to 4 by Using Recombinant Antigens

et al. 2002

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The B domains of dengue virus serotypes 1 to 4 were expressed in Escherichia coli. The purified proteins were applied to immunoblot strips to detect serotype-specific antibodies in paired serum samples from 41 patients with primary and secondary dengue infections. A close correlation between the results obtained with the immunoblot strips and by type-specific reverse transcription-PCR (T. Laue, P. Emmerich, and H. Schmitz, J. Clin. Microbiol. 37:2543-2547, 1999) was observed.

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Evaluation of recombinant dengue viral envelope B domain protein antigens for the detection of dengue complex-specific antibodies.

Cited by 31 publications

References 17 publications

Contribution of Disulfide Bridging to Epitope Expression of the Dengue Type 2 Virus Envelope Glycoprotein

Contribution of Disulfide Bridging to Epitope Expression of the Dengue Type 2 Virus Envelope Glycoprotein

Use of a Recombinant Envelope Protein Subunit Antigen for Specific Serological Diagnosis of West Nile Virus Infection

Serological Differentiation of Infections with Dengue Virus Serotypes 1 to 4 by Using Recombinant Antigens

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