Development of Envelope Protein Antigens To Serologically Differentiate Zika Virus Infection from Dengue Virus Infection

ABSTRACT Zika virus (ZIKV) is an emerging flavivirus that can cause birth defects and neurologic complications. Molecular tests are effective for diagnosing acute ZIKV infection, although the majority of infections produce no symptoms at all or present after the narrow window in which molecular diagnostics are dependable. Serology is a reliable method for detecting infections after the viremic period; however, most serological assays have limited specificity due to cross-reactive antibodies elicited by flavivirus infections. Since ZIKV and dengue virus (DENV) widely cocirculate, distinguishing ZIKV infection from DENV infection is particularly important for diagnosing individual cases or for surveillance to coordinate public health responses. Flaviviruses also elicit type-specific antibodies directed to non-cross-reactive epitopes of the infecting virus; such epitopes are attractive targets for the design of antigens for development of serological tests with greater specificity. Guided by comparative epitope modeling of the ZIKV envelope protein, we designed two recombinant antigens displaying unique antigenic regions on domain I (Z-EDI) and domain III (Z-EDIII) of the ZIKV envelope protein. Both the Z-EDI and Z-EDIII antigens consistently detected ZIKV-specific IgG in ZIKV-immune sera but not cross-reactive IgG in DENV-immune sera in late convalescence (>12 weeks postinfection). In contrast, during early convalescence (2 to 12 weeks postinfection), secondary DENV-immune sera and some primary DENV-immune sera cross-reacted with the Z-EDI and Z-EDIII antigens. Analysis of sequential samples from DENV-immune individuals demonstrated that Z-EDIII cross-reactivity peaked in early convalescence and declined steeply over time. The Z-EDIII antigen has much potential as a diagnostic antigen for population-level surveillance and for detecting past infections in patients.

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Ability To Serologically Confirm Recent Zika Virus Infection in Areas with Varying Past Incidence of Dengue Virus Infection in the United States and U.S. Territories in 2016

Journal of Clinical Microbiology ◽

10.1128/jcm.01115-17 ◽

2017 ◽

Vol 56 (1) ◽

Cited By ~ 13

Author(s):

Nicole P. Lindsey ◽

J. Erin Staples ◽

Krista Powell ◽

Ingrid B. Rabe ◽

Marc Fischer ◽

...

Keyword(s):

Puerto Rico ◽

Virus Infection ◽

Dengue Virus ◽

Zika Virus ◽

Denv Infection ◽

The United States ◽

American Samoa ◽

Cross Reactivity ◽

Immunoglobulin M ◽

Positive Results

ABSTRACTCross-reactivity within flavivirus antibody assays, produced by shared epitopes in the envelope proteins, can complicate the serological diagnosis of Zika virus (ZIKAV) infection. We assessed the utility of the plaque reduction neutralization test (PRNT) to confirm recent ZIKAV infections and rule out misleading positive immunoglobulin M (IgM) results in areas with various levels of past dengue virus (DENV) infection incidence. We reviewed PRNT results of sera collected for diagnosis of ZIKAV infection from 1 January through 31 August 2016 with positive ZIKAV IgM results, and ZIKAV and DENV PRNTs were performed. PRNT result interpretations included ZIKAV, unspecified flavivirus, DENV infection, or negative. For this analysis, ZIKAV IgM was considered false positive for samples interpreted as a DENV infection or negative. In U.S. states, 208 (27%) of 759 IgM-positive results were confirmed to be ZIKAV compared to 11 (21%) of 52 in the U.S. Virgin Islands (USVI), 15 (15%) of 103 in American Samoa, and 13 (11%) of 123 in Puerto Rico. In American Samoa and Puerto Rico, more than 80% of IgM-positive results were unspecified flavivirus infections. The false-positivity rate was 27% in U.S. states, 18% in the USVI, 2% in American Samoa, and 6% in Puerto Rico. In U.S. states, the PRNT provided a virus-specific diagnosis or ruled out infection in the majority of IgM-positive samples. Almost a third of ZIKAV IgM-positive results were not confirmed; therefore, providers and patients must understand that IgM results are preliminary. In territories with historically higher rates of DENV transmission, the PRNT usually could not differentiate between ZIKAV and DENV infections.

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Congenital abnormalities associated with Zika virus infection–Dengue as potential co-factor? A systematic review

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0008984 ◽

2021 ◽

Vol 15 (1) ◽

pp. e0008984

Author(s):

Stephanie Petzold ◽

Nisreen Agbaria ◽

Andreas Deckert ◽

Peter Dambach ◽

Volker Winkler ◽

...

Keyword(s):

Systematic Review ◽

Virus Infection ◽

Dengue Virus ◽

Protective Effect ◽

Zika Virus ◽

Congenital Abnormalities ◽

Denv Infection ◽

Current Evidence ◽

Geographic Variability ◽

Zikv Infection

Zika virus (ZIKV) emerged in Brazil during 2013–2014 causing an epidemic of previously unknown congenital abnormalities. The frequency of severe congenital abnormalities after maternal ZIKV infection revealed an unexplained geographic variability, especially between the Northeast and the rest of Brazil. Several reasons for this variability have been discussed. Prior immunity against DENV, that affects ZIKV seems to be the most likely explanation. Here we summarise the current evidence regarding the prominent co-factor to potentially explain the geographic variability. This systematic review followed the PRISMA guidelines. The search was conducted up to May 15th, 2020, focussing on immunological interactions from Zika virus with previous Dengue virus infections as potential teratogenic effect for the foetus. Eight out of 339 screened studies reported on the association between ZIKV, prior Dengue virus infection and microcephaly, mostly focusing on antibody-dependent enhancement (ADE) as potential pathomechanism. Prior DENV infection was associated with enhancement for ZIKV infection and increased neurovirulence in one included in vitro study only. Interestingly, the seven in vivo studies exhibited a heterogeneous picture with three studies showing a protective effect of prior DENV infections and others no effect at all. According to several studies, socio-economic factors are associated with increased risk for microcephaly. Very few studies addressed the question of unexplained variability of infection-related microcephaly. Many studies focussed on ADE as mechanism without measuring microcephaly as endpoint. Interestingly, three of the included studies reported a protective effect of prior DENV infection against microcephaly. This systematic review strengthens the hypothesis that immune priming after recent DENV infection is the crucial factor for determining protection or enhancement activity. It is of high importance that the currently ongoing prospective studies include a harmonized assessment of the potential candidate co-factors.

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Prior Dengue Virus Infection Is Associated With Increased Viral Load in Patients Infected With Dengue but Not Zika Virus

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz320 ◽

2019 ◽

Vol 6 (7) ◽

Cited By ~ 5

Author(s):

Gilberto A Santiago ◽

Tyler M Sharp ◽

Eli Rosenberg ◽

Iris I Sosa Cardona ◽

Luisa Alvarado ◽

...

Keyword(s):

Viral Load ◽

Virus Infection ◽

Dengue Virus ◽

Zika Virus ◽

Denv Infection ◽

T Test ◽

Viral Rna ◽

Zikv Infection ◽

Dengue Virus Infection

Abstract To evaluate potential enhancement of Zika virus (ZIKV) infection among patients with prior dengue virus (DENV) infection, we compared loads of viral RNA among patients infected with ZIKV (n = 1070), DENV-2 (n = 312), or DENV-3 (n = 260). Compared to patients without prior DENV infection, patients with prior DENV infection had significantly higher mean loads of viral RNA if infected with DENV-2 (10.6 vs 11.6 log10 GCE/mL, respectively; t test, P < .0001) or DENV-3 (10.3 vs 10.9 log10 GCE/mL; P < .0001), but not ZIKV (4.7 vs 4.7 log10 GCE/mL; P = .959). These findings provide evidence against in vivo enhancement of ZIKV by anti-DENV antibodies.

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Zika virus infection in a traveller returning from the Maldives, June 2015

Eurosurveillance ◽

10.2807/1560-7917.es.2016.21.2.30107 ◽

2016 ◽

Vol 21 (2) ◽

Cited By ~ 50

Author(s):

Essi Marjana Korhonen ◽

Eili Huhtamo ◽

Teemu Smura ◽

Hannimari Kallio-Kokko ◽

Markku Raassina ◽

...

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Zika Virus ◽

Denv Infection ◽

Distribution Range ◽

Rt Pcr ◽

Zikv Infection ◽

Igm Antibodies ◽

Zika Virus Infection

We report a Zika virus (ZIKV) infection in a patient with fever and rash after returning to Finland from Maldives, June 2015. The patient had dengue virus (DENV) IgG and IgM antibodies but pan-flavivirus RT-PCR and subsequent sequencing showed presence of ZIKV RNA in urine. Recent association of ZIKV with microcephaly highlights the need for laboratory differentiation of ZIKV from DENV infection and the circulation of ZIKV in areas outside its currently known distribution range.

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Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

Clinical Infectious Diseases ◽

10.1093/cid/cix672 ◽

2017 ◽

Vol 65 (11) ◽

pp. 1829-1836 ◽

Cited By ~ 40

Author(s):

Wen-Yang Tsai ◽

Han Ha Youn ◽

Carlos Brites ◽

Jih-Jin Tsai ◽

Jasmine Tyson ◽

...

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Zika Virus ◽

Dengue Virus Infection ◽

Serological Tests ◽

Zika Virus Infection

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Low seroprevalence of Zika virus infection among adults in Southern Taiwan

BMC Infectious Diseases ◽

10.1186/s12879-019-4491-4 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 2

Author(s):

Yu-Wen Chien ◽

Tzu-Chuan Ho ◽

Pei-Wen Huang ◽

Nai-Ying Ko ◽

Wen-Chien Ko ◽

...

Keyword(s):

Dengue Virus ◽

Migrant Workers ◽

Zika Virus ◽

Southeast Asian ◽

Asian Countries ◽

Zikv Infection ◽

Serological Tests ◽

Cross Sectional ◽

Denv Serotypes ◽

Southeast Asian Countries

Abstract Background We recently conducted a serosurvey of newly arrived workers in Taiwan from four Southeast Asian countries which revealed that 1% of the migrant workers had laboratory-confirmed recent Zika virus (ZIKV) infection. Taiwan, where Aedes mosquitoes are prevalent, has a close relationship with Southeast Asian countries. Up to now, 21 imported cases of ZIKV infection have been reported in Taiwan, but there has been no confirmed indigenous case. The aim of this serosurvey was to assess whether there was unrecognized ZIKV infections in Taiwan. Methods A total of 212 serum samples collected in a cross-sectional seroepidemiologic study conducted during the end of the 2015 dengue epidemic in Tainan, Taiwan, were analyzed. Anti-ZIKV IgM and IgG were tested using commercial enzyme-linked immunosorbent assays (ELISAs). Plaque reduction neutralization tests (PRNTs) for ZIKV and four dengue virus (DENV) serotypes were performed for samples with positive anti-ZIKV antibodies. A confirmed case of ZIKV infection was defined by ZIKV PRNT90 titer ratio ≥ 4 compared to four DENV serotypes. Results The mean age of the 212 participants was 54.0 years (standard deviation 13.7 years), and female was predominant (67.0%). Anti-ZIKV IgM and IgG were detected in 0 (0%) and 9 (4.2%) of the 212 participants, respectively. For the 9 samples with anti-ZIKV IgG, only 1 sample had 4 times higher ZIKV PRNT90 titers compared to PRNT90 titers against four dengue virus serotypes; this individual denied having traveled abroad. Conclusions The results suggest that undetected indigenous ZIKV transmission might have occurred in Taiwan. The findings also suggest that the threat of epidemic transmission of ZIKV in Taiwan does exist due to extremely low-level of herd immunity. Our study also indicates that serological tests for ZIKV-specific IgG remain a big challenge due to cross-reactivity, even in dengue non-endemic countries.

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Dengue virus infection impedes megakaryopoiesis in MEG-01 cells where the virus envelope protein interacts with the transcription factor TAL-1

Scientific Reports ◽

10.1038/s41598-020-76350-5 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Atoshi Banerjee ◽

Aarti Tripathi ◽

Shweta Duggal ◽

Arup Banerjee ◽

Sudhanshu Vrati

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Envelope Protein ◽

Marked Reduction ◽

Leukemia Cell ◽

Denv Infection ◽

Host Protein ◽

Leukemia Cell Line ◽

Virus Envelope ◽

Virus Envelope Protein

Abstract Dengue virus (DENV) infection causes dengue fever in humans, which can lead to thrombocytopenia showing a marked reduction in platelet counts, and dengue hemorrhagic fever. The virus may cause thrombocytopenia either by destroying the platelets or by interfering with their generation via the process of megakaryopoiesis. MEG-01 is the human megakaryoblastic leukemia cell line that can be differentiated in vitro by phorbol-12-myristate-13-acetate (PMA) treatment to produce platelet-like-particles (PLPs). We have studied DENV infection of MEG-01 cells to understand its effect on megakaryopoiesis and the generation of PLPs. We observed that DENV could infect only naive MEG-01 cells, and differentiated cells were refractory to virus infection/replication. However, DENV-infected MEG-01 cells, when induced for differentiation with PMA, supported an enhanced viral replication. Following the virus infection, the MEG-01 cells showed a marked reduction in the surface expression of platelet markers (CD41, CD42a, and CD61), a decreased polyploidy, and significantly reduced PLP counts. DENV infection caused an enhanced Notch signaling in MEG-01 cells where the virus envelope protein was shown to interact with TAL-1, a host protein important for megakaryopoiesis. These observations provide new insight into the role of DENV in modulating the megakaryopoiesis and platelet production process.

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Diagnostic performance of anti-Zika virus IgM, IgAM and IgG ELISAs during co-circulation of Zika, dengue, and chikungunya viruses in Brazil and Venezuela

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009336 ◽

2021 ◽

Vol 15 (4) ◽

pp. e0009336

Author(s):

Ivonne Morales ◽

Kerstin D. Rosenberger ◽

Tereza Magalhaes ◽

Clarice N. L. Morais ◽

Cynthia Braga ◽

...

Keyword(s):

Zika Virus ◽

Test Performance ◽

Diagnostic Performance ◽

Cross Reactivity ◽

Rt Pcr ◽

Zikv Infection ◽

Serological Tests ◽

Sequential Samples

Background Serological diagnosis of Zika virus (ZIKV) infection is challenging because of the antibody cross-reactivity among flaviviruses. At the same time, the role of Nucleic Acid Testing (NAT) is limited by the low proportion of symptomatic infections and the low average viral load. Here, we compared the diagnostic performance of commercially available IgM, IgAM, and IgG ELISAs in sequential samples during the ZIKV and chikungunya (CHIKV) epidemics and co-circulation of dengue virus (DENV) in Brazil and Venezuela. Methodology/Principal findings Acute (day of illness 1–5) and follow-up (day of illness ≥ 6) blood samples were collected from nine hundred and seven symptomatic patients enrolled in a prospective multicenter study of symptomatic patients recruited between June 2012 and August 2016. Acute samples were tested by RT-PCR for ZIKV, DENV, and CHIKV. Acute and follow-up samples were tested for IgM, IgAM, and IgG antibodies to ZIKV using commercially available ELISAs. Among follow-up samples with a RT-PCR confirmed ZIKV infection, anti-ZIKV IgAM sensitivity was 93.5% (43/48), while IgM and IgG exhibited sensitivities of 30.3% (10/35) and 72% (18/25), respectively. An additional 24% (26/109) of ZIKV infections were detected via IgAM seroconversion in ZIKV/DENV/CHIKV RT-PCR negative patients. The specificity of anti-ZIKV IgM was estimated at 93% and that of IgAM at 85%. Conclusions/Significance Our findings exemplify the challenges of the assessment of test performance for ZIKV serological tests in the real-world setting, during co-circulation of DENV, ZIKV, and CHIKV. However, we can also demonstrate that the IgAM immunoassay exhibits superior sensitivity to detect ZIKV RT-PCR confirmed infections compared to IgG and IgM immunoassays. The IgAM assay also proves to be promising for detection of anti-ZIKV seroconversions in sequential samples, both in ZIKV PCR-positive as well as PCR-negative patients, making this a candidate assay for serological monitoring of pregnant women in future ZIKV outbreaks.

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T-cell Responses in Individuals Infected with Zika Virus and in Those Vaccinated Against Dengue Virus

Pathogens and Immunity ◽

10.20411/pai.v2i2.188 ◽

2017 ◽

Vol 2 (2) ◽

pp. 274 ◽

Cited By ~ 13

Author(s):

Dominic Paquin-Proulx ◽

Fabio E. Leal ◽

Cassia G. Terrassani Silveira ◽

Alvino Maestri ◽

Claudia Brockmeyer ◽

...

Keyword(s):

T Cell ◽

Dengue Virus ◽

Zika Virus ◽

Cell Response ◽

Cell Epitope ◽

Denv Infection ◽

Cross Reactivity ◽

T Cell Response ◽

Cd4 T Cell ◽

T Cell Epitopes

Background: The outbreak of Zika virus (ZIKV) infection in Brazil has raised concerns that infection during pregnancy could cause microcephaly and other severe neurodevelopmental malformations in the fetus. The mechanisms by which ZIKV causes fetal abnormalities are largely unknown. The importance of pre-infection with dengue virus (DENV), or other flaviviruses endemic to Brazil, remains to be investigated. It has been reported that antibodies directed against DENV can increase ZIKV infectivity by antibody dependent enhancement (ADE), suggesting that a history of prior DENV infection might worsen the outcome of ZIKV infection.Methods: We used bioinformatics tools to design 18 peptides from the ZIKV envelope containing predicted HLA-I T-cell epitopes and investigated T-cell cross-reactivity between ZIKV-infected individuals and DENV-vaccinated subjects by IFNg ELISPOT.Results: Three peptides induced IFNg production in both ZIKV-infected subjects and in DENV-vaccinated individuals. Flow cytometry indicated that 1 ZIKV peptide induced a CD4+ T-cell response in DENV-vaccinated subjects.Conclusions: We demonstrated that vaccination against DENV induced a T-cell response against ZIKV and identified one such CD4+ T-cell epitope. The ZIKV-reactive CD4+ T cells induced by DENV vaccination and identified in this study could contribute to the appearance of cross-reactive antibodies mediating ADE.

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Global transcriptome analysis of Aedes aegypti mosquitoes in response to Zika virus infection

10.1101/179416 ◽

2017 ◽

Cited By ~ 1

Author(s):

Kayvan Etebari ◽

Shivanand Hegde ◽

Miguel A Saldaña ◽

Steven G Widen ◽

Thomas G Wood ◽

...

Keyword(s):

Virus Infection ◽

Aedes Aegypti ◽

Dengue Virus ◽

Zika Virus ◽

Target Genes ◽

World Health ◽

Mosquito Vector ◽

Zikv Infection ◽

Zika Virus Infection ◽

Non Coding Rnas

AbstractZika virus (ZIKV) of the Flaviviridae family is a recently emerged mosquito-borne virus that has been implicated in the surge of the number of microcephaly instances in South America. The recent rapid spread of the virus led to its declaration as a global health emergency by the World Health Organization. The virus is transmitted mainly by the mosquito Aedes aegypti that also vectors dengue virus, however little is known about the interactions of the virus with the mosquito vector. In this study, we investigated the transcriptome profiles of whole Ae. aegypti mosquitoes in response to ZIKV infection at 2, 7, and 14 days post-infection using RNA-Seq. Results showed changes in the abundance of a large number of transcripts at each time point following infection, with 18 transcripts commonly changed among the three time points. Gene ontology analysis revealed that most of the altered genes are involved in metabolic process, cellular process and proteolysis. In addition, 486 long intergenic non-coding RNAs were identified that were altered upon ZIKV infection. Further, we found correlational changes of a number of potential mRNA target genes with that of altered host microRNAs. The outcomes provide a basic understanding of Ae. aegypti responses to ZIKV and helps to determine host factors involved in replication or mosquito host anti-viral response against the virus.ImportanceVector-borne viruses pose great risks on human health. Zika virus has recently emerged as a global threat, rapidly expanding its distribution. Understanding the interactions of the virus with mosquito vectors at the molecular level is vital for devising new approaches in inhibiting virus transmission. In this study, we embarked on analyzing the transcriptional response of Aedes aegypti mosquitoes to Zika virus infection. Results showed large changes both in coding and long non-coding RNAs. Analysis of these genes showed similarities with other flaviviruses, including dengue virus, which is transmitted by the same mosquito vector. The outcomes provide a global picture of changes in the mosquito vector in response to Zika virus infection.

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