Human immune response to Puumala virus glycoproteins and nucleocapsid protein expressed in mammalian cells

Kallio‐Kokko, Hannimari; Leveelahti, Raija; Brummer-Korvenkontio, M; Lundkvist, Åke; Vaheri, Antti; Vapalahti, Olli

doi:10.1002/jmv.2079

Cited by 46 publications

(31 citation statements)

References 40 publications

(41 reference statements)

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“…The IgM test (data not utilized in the present paper) was an IgM capture enzyme immunoassay based on recombinant PUUV-N produced in the baculovirus system [32]. The IgG test was an in-house immunofluorescence assay (IFA) based on PUUV-infected acetone-fixed Vero E6 cells [33,34]. …”

Section: Methodsmentioning

confidence: 99%

Serogrouping and seroepidemiology of North European hantaviruses using a novel broadly targeted synthetic nucleoprotein antigen array

Rönnberg

Vapalahti

Goeijenbier

et al. 2017

Infection Ecology & Epidemiology

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Introduction: Hantaviruses are globally distributed zoonotic pathogens. Great diversity and high antigenic cross-reactivity makes diagnosis by traditional methods cumbersome.Materials and methods: ‘Megapeptides’, 119–120-mers from the amino terminus of the nucleoprotein of 16 hantaviruses, representing the four major branches of the hantavirus phylogenetic tree, were utilized in a novel IgG-based hantavirus suspension multiplex immunoassay (HSMIA) for detection of past hantavirus infections in 155 North European human samples. We compared HSMIA with established EIAs and focus reduction neutralization test (FRNT).Results and discussion: The Puumala hantavirus (PUUV) component in the HSMIA gave concordant results with a PUUV IgG EIA in 142 sera from Northern Sweden (of which 31 were EIA positive, 7 borderline and 104 EIA negative, sensitivity 30/31 = 97%, specificity 104/ 104 = 100%, 134/135 = 99% concordance), with another immunoassay in 40 PUUV IgG positive sera from Finland (36/40 = 90% sensitivity), and was concordant in 8 of 11 cases with PUUV and DOBV neutralization titers, respectively. Two major IgG reactivity patterns were found: (i) a PUUV-specific pattern covering phylogroup IV and its serogroups B and C; and (ii) a Dobrava virus (DOBV)-specific pattern, covering the serogroup A portion of phylogroup III. In addition, we found several minor patterns with reactivity to only one or two megapeptides indicating additional hantaviruses infecting humans in the Swedish and Finnish populations.Conclusion: The broadly reactive and rational HSMIA yielded results highly correlated with the established PUUV EIAs and the NT results. It is a sensitive and specific assay, which will be suited for efficient serosurveillance of hantaviruses in humans. Its use in animals should be further investigated.

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Section: Methodsmentioning

confidence: 99%

Serogrouping and seroepidemiology of North European hantaviruses using a novel broadly targeted synthetic nucleoprotein antigen array

Rönnberg

Vapalahti

Goeijenbier

et al. 2017

Infection Ecology & Epidemiology

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“…The IgM test is an IgM capture enzyme immunoassay based on recombinant PUUV-N produced in the baculovirus system similarly to the antigen we use in the LFRET assay (21). The IgG test is an in-house immunofluorescence assay (IFA) based on PUUV-infected acetone-fixed Vero E6 cells (9,10,21 CH1-b assay. Protein L targets the light chain and recognizes ϳ50% of Ig molecules of nearly all isotypes.…”

Section: Methodsmentioning

confidence: 99%

“…Hantavirus infection induces a strong humoral IgM and IgG antibody response against the structural proteins, particularly the nucleocapsid (N) protein in the acute phase. Laboratory diagnosis of hantavirus disease is based on the detection of IgM and IgG antibodies, usually by immunochromatography (IgM), enzyme immunoassay (EIA) with recombinant N as the antigen, or immunofluorescence assay (IFA) based on acetone-fixed infected cells (9)(10)(11).…”

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confidence: 99%

Rapid Homogeneous Immunoassay Based on Time-Resolved Förster Resonance Energy Transfer for Serodiagnosis of Acute Hantavirus Infection

Hepojoki

Hedman

et al. 2015

J Clin Microbiol

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We recently introduced a homogeneous immunoassay based on time-resolved Förster resonance energy transfer (TR-FRETP uumala virus (PUUV) belongs to the genus Hantavirus within the family Bunyaviridae, comprising typically rodent-and insectivore-borne negative-stranded RNA viruses (1). The known reservoir hosts of hantaviruses are chronically infected and spread the virus via excreta (saliva, urine, and feces) (2). The human infection, acquired via inhalation of aerosolized rodent excreta, manifests as two disease entities: hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). HFRS predominates in Eurasia, and HCPS occurs in the Americas. The case fatality rate of HFRS is 0.1 to 12%, depending on the virus type, and that of HCPS is up to 40% (3).Annually, around 50,000 hospitalizations due to HFRS occur in Eurasia, while a total of ϳ3,500 HCPS cases have been recorded since 1993 (4). In Europe, the most common cause of HFRS (up to 10,000 cases per annum) is PUUV that circulates in bank voles (Myodes glareolus) (5). The symptoms of PUUV infection, nephropathia epidemica (NE, a mild form of HFRS), include abrupt fever, followed by headache, nausea, vomiting, abdominal pain, and signs of renal insufficiency; somnolence and visual disturbances are common, and pulmonary, cardiac, and central nervous system symptoms have also been recorded. The incubation period is 2 to 6 weeks (5-8). Hantavirus infection induces a strong humoral IgM and IgG antibody response against the structural proteins, particularly the nucleocapsid (N) protein in the acute phase. Laboratory diagnosis of hantavirus disease is based on the detection of IgM and IgG antibodies, usually by immunochromatography (IgM), enzyme immunoassay (EIA) with recombinant N as the antigen, or immunofluorescence assay (IFA) based on acetone-fixed infected cells (9-11).Förster resonance energy transfer (FRET) has been applied widely in biomedical research (12)(13)(14)(15)(16)(17)(18)(19), and our goal is to harness the phenomenon for infectious disease serodiagnostics. In FRET, an excited donor fluorophore transfers its energy to an acceptor fluorophore in close proximity (Ͻ10 nm). Time-resolved FRET (TR-FRET) utilizes the long fluorescence emission half-lives of lanthanides and enables background-free measurement from biological materials. We recently introduced two different principles utilizing TR-FRET for antibody detection. The first approach is based on binding of an immunoglobulin molecule to its antigens labeled distinctly with donor and acceptor fluorophores, giving rise to a FRET signal in a homogenous (wash-free) assay format (12). The second approach is based on binding of an immunoglobulin molecule to its antigen and protein L (a bacterial protein targeting the Ig kappa light chain) labeled distinctly with fluorophores forming a FRET pair (20). The emitted TR-FRET signal is proportional to the amount of specific immunoglobulin. Here, using recombinant full-length N protein of PUUV (PUUV-N), we showed that the protein...

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“…The reference test results for PUUV IgM (specificity, 100%; sensitivity, 100%) and IgG (specificity, 100%; sensitivity, 96%) antibodies were obtained from HUSLAB (7,19,(21)(22)(23). The IgG antibodies were detected by an in-house immunofluorescence assay (IFA) based on PUUV-infected acetone-fixed Vero E6 cells (6,7,19). The IgM antibodies against PUUV-N were detected by -capture IgM EIA based on recombinant PUUV-N produced as for the CFRET-IA (19).…”

Section: Methodsmentioning

confidence: 99%

“…The antibody detection is commonly based on enzyme immunoassay (EIA), immunoblotting using recombinant PUUV N as antigen, or immunofluorescence assay (IFA) based on acetone-fixed infected cells. Additionally, rapid immunochromatography tests for IgM detection are available (6)(7)(8).…”

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confidence: 99%

Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease

et al. 2015

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In this study, we describe a competitive homogeneous immunoassay that makes use of Förster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease.

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Human immune response to Puumala virus glycoproteins and nucleocapsid protein expressed in mammalian cells

Cited by 46 publications

References 40 publications

Serogrouping and seroepidemiology of North European hantaviruses using a novel broadly targeted synthetic nucleoprotein antigen array

Serogrouping and seroepidemiology of North European hantaviruses using a novel broadly targeted synthetic nucleoprotein antigen array

Rapid Homogeneous Immunoassay Based on Time-Resolved Förster Resonance Energy Transfer for Serodiagnosis of Acute Hantavirus Infection

Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease

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