A neutralization assay for chikungunya virus infections in a multiplex format

BackgroundOutbreaks of the Chikungunya virus (CHIKV) infection has been documented in over 40 countries, resulting in clinical symptoms characterized by fever and joint pain. Diagnosing CHIKV in a clinical lab setting is often omitted because of the high lab safety requirement. An infection system that mimics CHIKV infection will permit clinical evaluation of the production of neutralizing antibody for both disease diagnostics and treatment.Methodology/Principal FindingsWe generated a CHIKV construct expressing CHIKV structural proteins. This construct permits the production of CHIKV pseudo-viral particles with a luciferase reporter. The pseudo-virus was able to infect a wide range of cell lines. The pseudovirus could be neutralized by the addition of neutralizing antibodies from patients.ConclusionsTaken together, we have developed a powerful system that can be handled at biosafety level 2 laboratories for evaluation of existence of CHIKV neutralizing antibodies.

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“…It also allows the study of CHIKV entry. Our method is similar to what has been reported [12] , [13] , [14] .…”

Section: Discussionsupporting

confidence: 82%

Chikungunya Virus Glycoproteins Pseudotype with Lentiviral Vectors and Reveal a Broad Spectrum of Cellular Tropism

Zhang

Wang

et al. 2014

PLoS ONE

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“…Slides were washed and incubated with FITC-labeled rabbit anti-human IgG (SIFIN, Berlin, Germany) (Schwarz et al 2012). The neutralization test was performed with a lentiviral vector that was pseudotyped with the CHIKV glycoprotein (Weber et al 2014).…”

mentioning

confidence: 99%

Low Seroprevalence Indicates Vulnerability of Eastern and Central Sudan to Infection with Chikungunya Virus

Adam¹,

Seidahmed

Weber

et al. 2016

Vector-Borne and Zoonotic Diseases

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Outbreaks of infections with chikungunya virus (CHIKV) have previously been reported from Sudan but the prevalence in the general population is unknown. We investigated the seroprevalence of CHIKV infection in 379 serum samples from patients with fever in the outpatient clinics of three hospitals in eastern and central Sudan. The seroprevalence was 1.8%, indicating that CHIKV infections are rare in these parts of Sudan. As the vector Aedes aegypti is endemic in this area, the population is at risk for a CHIKV epidemic.

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“…The codon-optimized CHIKV E3-E1 gene (based on isolate “S27”) was synthesized by GeneArt (Life Technologies, Darmstadt) and cloned into the plasmid pIRES2-eGFP (Clontech/Takara, 78100 Saint-Germain-en-Laye, France) as described previously [ 18 ].…”

Section: Methodsmentioning

confidence: 99%

A Small Antigenic Determinant of the Chikungunya Virus E2 Protein Is Sufficient to Induce Neutralizing Antibodies which Are Partially Protective in Mice

2015

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BackgroundThe mosquito-borne Chikungunya virus (CHIKV) causes high fever and severe joint pain in humans. It is expected to spread in the future to Europe and has recently reached the USA due to globalization, climate change and vector switch. Despite this, little is known about the virus life cycle and, so far, there is no specific treatment or vaccination against Chikungunya infections. We aimed here to identify small antigenic determinants of the CHIKV E2 protein able to induce neutralizing immune responses.Methodology/Principal FindingsE2 enables attachment of the virus to target cells and a humoral immune response against E2 should protect from CHIKV infections. Seven recombinant proteins derived from E2 and consisting of linear and/or structural antigens were created, and were expressed in and purified from E. coli. BALB/c mice were vaccinated with these recombinant proteins and the mouse sera were screened for neutralizing antibodies. Whereas a linear N-terminally exposed peptide (L) and surface-exposed parts of the E2 domain A (sA) alone did not induce neutralizing antibodies, a construct containing domain B and a part of the β-ribbon (called B+) was sufficient to induce neutralizing antibodies. Furthermore, domain sA fused to B+ (sAB+) induced the highest amount of neutralizing antibodies. Therefore, the construct sAB+ was used to generate a recombinant modified vaccinia virus Ankara (MVA), MVA-CHIKV-sAB+. Mice were vaccinated with MVA-CHIKV-sAB+ and/or the recombinant protein sAB+ and were subsequently challenged with wild-type CHIKV. Whereas four vaccinations with MVA-CHIKV-sAB+ were not sufficient to protect mice from a CHIKV infection, protein vaccination with sAB+ markedly reduced the viral titers of vaccinated mice.Conclusions/SignificanceThe recombinant protein sAB+ contains important structural antigens for a neutralizing antibody response in mice and its formulation with appropriate adjuvants might lead to a future CHIKV vaccine.

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A neutralization assay for chikungunya virus infections in a multiplex format

Cited by 25 publications

References 25 publications

Chikungunya Virus Glycoproteins Pseudotype with Lentiviral Vectors and Reveal a Broad Spectrum of Cellular Tropism

Chikungunya Virus Glycoproteins Pseudotype with Lentiviral Vectors and Reveal a Broad Spectrum of Cellular Tropism

Low Seroprevalence Indicates Vulnerability of Eastern and Central Sudan to Infection with Chikungunya Virus

A Small Antigenic Determinant of the Chikungunya Virus E2 Protein Is Sufficient to Induce Neutralizing Antibodies which Are Partially Protective in Mice

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