scholarly journals Alanine Substitution Inactivates Cross-Reacting Epitopes in Dengue Virus Recombinant Envelope Proteins

Viruses ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 208
Author(s):  
Viviana C. Zomosa-Signoret ◽  
Karina R. Morales-González ◽  
Ana E. Estrada-Rodríguez ◽  
Ana M. Rivas-Estilla ◽  
M. Cristina Devèze-García ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Rural Areas ◽  
Cross Reactivity ◽  
Diagnostic Methods ◽  
Hemorrhagic Complication ◽  
Virus Envelope ◽  
Recombinant Peptide ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Dengue Diagnostics

The expansion of the habitat of mosquitoes belonging to the Aedes genus puts nearly half of the world’s population at risk of contracting dengue fever, and a significant fraction will develop its serious hemorrhagic complication, which can be fatal if not diagnosed properly and treated in a timely fashion. Although several diagnostic methods have been approved for dengue diagnostics, their applicability is limited in rural areas of developing countries by sample preparation costs and methodological requirements, as well as cross-reactivity among the different serotypes of the Dengue virus and other flavivirus, such as the Zika virus. For these reasons, it is necessary to generate more specific antigens to improve serological methods that could be cheaper and used in field operations. Here, we describe a strategy for the inactivation of cross-reacting epitopes on the surface of the Dengue virus envelope protein through the synthetic generation of recombinant peptide sequences, where key amino acid residues from Dengue virus serotype 1 (DENV-1) and 2 (DENV-2) are substituted by alanine residues. The proteins thus generated are recognized by 88% of sera from Dengue NS1+ patients and show improved serotype specificity because they do not react with the antibodies present in seroconverted, PCR-serotyped DEN-4 infected patients.

scholarly journals Tracking the polyclonal neutralizing antibody response to a dengue virus serotype 1 type-specific epitope across two populations in Asia and the Americas

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Daniela V. Andrade ◽  
Colin Warnes ◽  
Ellen Young ◽  
Leah C. Katzelnick ◽  
Angel Balmaseda ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Sri Lankan ◽  
Human Monoclonal Antibodies ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Dengue Virus Serotypes ◽  
Kinetics Of ◽  
Two Populations

Abstract The four dengue virus serotypes (DENV1-4) cause major public health problems worldwide. Highly neutralizing type-specific human monoclonal antibodies (hmAbs) target conformation-dependent epitopes on the DENV envelope protein, including 1F4, a DENV1 type-specific hmAb. Using a recombinant DENV2 virus displaying the DENV1 1F4 epitope (rDENV2/1), we measured the proportion and kinetics of DENV1 neutralizing antibodies targeting the 1F4 epitope in individuals living in Asia and the Americas where different DENV1 genotypes were circulating. Samples from 20 individuals were analyzed 3 and 18 months post-primary DENV1 infection, alongside samples from 4 individuals collected annually for four years post-primary DENV1 infection, from two studies in Nicaragua. We also analyzed convalescent post-primary DENV1 plasma samples from Sri Lankan individuals. We found that neutralizing antibodies recognizing the 1F4 epitope vary in prevalence across both populations and were detected from 20 days to four years post-infection. Additionally, both populations displayed substantial variability, with a range of high to low proportions of DENV1 type-specific neutralizing antibodies recognizing the 1F4 epitope seen across individuals. Thus, the 1F4 epitope is a major but not exclusive target of type-specific neutralizing antibodies post-primary infection with different DENV1 genotypes in Asia and Latin America, and additional epitopes likely contribute to type-specific neutralization of DENV1.

scholarly journals Enhanced Prokaryotic Expression of Dengue Virus Envelope Protein

2013 ◽  
Vol 16 (4) ◽  
pp. 609 ◽  
Author(s):  
Advaita Ganguly ◽  
Ravindra B. Malabadi ◽  
Dipankar Das ◽  
Mavanur R. Suresh ◽  
Hoon H Sunwoo
Keyword(s):  
Dengue Virus ◽  
Envelope Protein ◽  
Vaccine Development ◽  
Virus Envelope ◽  
E Coli ◽  
C Terminus ◽  
Virus Envelope Protein ◽  
Biological Functionality ◽  
Dengue Diagnostics

Purpose. To highlight the expression and purification of the recombinant dengue virus type-1 antigen exploiting the codon optimized full length envelope for increased yield in E. coli. Methods. A 6x His tag was inserted at the C terminus to facilitate purification. The purified protein was recognized in Western blot by Monoclonal antibody specific for the tag. The in vitro refolded recombinant protein was used to immunize mice for the development of hybridomas and also analyzed for its biological functionality with heparan sulfate binding assay. Results. The polyclonal anti-sera from the immunized mice were found to recognize the envelope protein thereby establishing the immunogenicity of the protein. Conclusion. The purified envelope protein could potentially be used towards dengue diagnostics and vaccine development efforts. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

scholarly journals Genotypic Differences in Dengue Virus Neutralization Are Explained by a Single Amino Acid Mutation That Modulates Virus Breathing

mBio ◽  
2015 ◽  
Vol 6 (6) ◽  
Author(s):  
Kimberly A. Dowd ◽  
Christina R. DeMaso ◽  
Theodore C. Pierson
Keyword(s):  
Amino Acid ◽  
Dengue Virus ◽  
Vaccine Development ◽  
Antibody Binding ◽  
Single Amino Acid ◽  
Genotypic Differences ◽  
Conformational Ensembles ◽  
Single Amino Acid Residue ◽  
Virus Serotype ◽  
Serotype 1

ABSTRACTFlaviviruses sample an ensemble of virion conformations resulting from the conformational flexibility of their structural proteins. To investigate how sequence variation among strains impacts virus breathing, we performed studies with the monoclonal antibody (MAb) E111, which binds an inaccessible domain III envelope (E) protein epitope of dengue virus serotype 1 (DENV1). Prior studies indicated that an observed ~200-fold difference in neutralization between the DENV1 strains Western Pacific-74 (West Pac-74) and 16007 could not be explained by differences in the affinity of MAb E111 for each strain. Through neutralization studies with wild-type and variant viruses carrying genes encoding reciprocal mutations at all 13 amino acid differences between the E proteins of West Pac-74 and 16007, we found that E111 neutralization susceptibility mapped solely to the presence of a lysine or arginine at E domain II residue 204, located distally from the E111 epitope. This same residue correlated with neutralization differences observed for MAbs specific for epitopes distinct from E111, suggesting that this amino acid dictates changes in the conformational ensembles sampled by the virus. Furthermore, an observed twofold difference in the stability of infectious West Pac-74 versus 16007 in solution also mapped to E residue 204. Our results demonstrate that neutralization susceptibility can be altered in an epitope-independent manner by natural strain variation that influences the structures sampled by DENV. That different conformational ensembles of flaviviruses may affect the landscape available for antibody binding, as well as virus stability, has important implications for functional studies of antibody potency, a critical aspect of vaccine development.IMPORTANCEThe global burden of dengue virus (DENV) is growing, with recent estimates of ~390 million human infections each year. Antibodies play a crucial role in protection from DENV infection, and vaccines that elicit a robust antibody response are being actively pursued. We report here the identification of a single amino acid residue in the envelope protein of DENV serotype 1 that results in global changes to virus structure and stability when it is changed. Our results indicate that naturally occurring variation at this particular site among virus strains impacts the ensemble of structures sampled by the virus, a process referred to as virus breathing. The finding that such limited and conservative sequence changes can modulate the landscape available for antibody binding has important implications for both vaccine development and the study of DENV-reactive antibodies.

Induction of multiple cytotoxic T lymphocyte responses in mice by a multiepitope DNA vaccine against dengue virus serotype 1

2016 ◽  
Vol 60 (12) ◽  
pp. 835-845 ◽  
Author(s):  
Xin Yu Chen ◽  
De Zhou Li ◽  
Xiao Zhi Zhong ◽  
Bokun Chen ◽  
Zhi Liang Duan ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dna Vaccine ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Lymphocyte Responses ◽  
Dengue Virus Serotype 1

scholarly journals Safety and Immunogenicity of a Dengue Virus Serotype-1 Purified-Inactivated Vaccine: Results of a Phase 1 Clinical Trial

2015 ◽  
Vol 93 (3) ◽  
pp. 454-460 ◽  
Author(s):  
Luis Javier Martinez ◽  
Leyi Lin ◽  
Jason M. Blaylock ◽  
Arthur G. Lyons ◽  
Kristen M. Bauer ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Dengue Virus ◽  
Phase 1 ◽  
Inactivated Vaccine ◽  
Phase 1 Clinical Trial ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Dengue Virus Serotype 1

scholarly journals Rapid Subtyping of Dengue Virus Serotypes 1 and 4 by Restriction Site-Specific PCR

2000 ◽  
Vol 38 (3) ◽  
pp. 1286-1289 ◽  
Author(s):  
Marize P. Miagostovich ◽  
Flavia B. dos Santos ◽  
C. Milena Gutiérrez ◽  
Lee W. Riley ◽  
Eva Harris
Keyword(s):  
Sequence Analysis ◽  
Dengue Virus ◽  
Restriction Site ◽  
Epidemiological Studies ◽  
Site Specific ◽  
Specific Pcr ◽  
Virus Serotype ◽  
Serotype 1 ◽  
Dengue Virus Serotypes ◽  
Dengue Virus Serotype 1

We previously reported a simple subtyping method, restriction site-specific PCR (RSS-PCR), for dengue virus serotypes 2 and 3; here we describe its application for subtyping dengue virus serotypes 1 and 4. Three major RSS-PCR types were observed for dengue virus serotype 1 and two types were observed for dengue virus serotype 4, in agreement with previous strain classifications based on sequence analysis. Because of its simplicity, this method is amenable to rapid subtyping and application to epidemiological studies of dengue in countries where dengue is endemic.

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