Dengue virus isolation relying on antibody-dependent enhancement mechanism using FcγR-expressing BHK cells and a monoclonal antibody with infection-enhancing capacity

Moi, Meng Ling; Lim, Chang‐Kweng; Tajima, Shigeru; Kotaki, Akira; Saijo, Masayuki; Takasaki, Tomohiko; Kurane, Ichiro

doi:10.1016/j.jcv.2011.07.009

Cited by 6 publications

(3 citation statements)

References 18 publications

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“…The results were in agreement with other findings: all ten sera from dengue patients with secondary infections enhanced antibody-dependent DENV infection of K562 cells. As seen in other studies [23], [38], [47], we also found that there was no correlation between in vitro ADE activity and disease severity. In the present study, only one out of ten serum samples (HTD45) was incapable of virus neutralization.…”

Section: Discussionsupporting

confidence: 86%

“…Previous studies assayed ADE in K562 [10], [26], [34], THP-1 [23], [35]–[37] or BHK [38], [39] cells. Therefore, we used both THP-1 and K562 cells to assay ADE (Supplemental Figure S5).…”

Section: Resultsmentioning

confidence: 99%

“…Some studies used clinical isolates prepared from dengue patient plasma, which had been passaged in cell lines [38], [48]. Therefore, we used laboratory-grown DENV with a serotype identical to that of the serotype detected in patient plasma.…”

Section: Discussionmentioning

confidence: 99%

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Low Levels of Antibody-Dependent Enhancement in Vitro Using Viruses and Plasma from Dengue Patients

et al. 2014

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BackgroundThe majority of dengue patients infected with any serotype of dengue virus (DENV) are asymptomatic, but the remainder may develop a wide spectrum of clinical symptoms, ranging from mild dengue fever (DF) to severe dengue hemorrhagic fever (DHF). Severe cases occur more often in patients who experience a secondary infection with a different virus serotype. A phenomenon called antibody-dependent enhancement (ADE) has been proposed to explain the onset of these severe cases, but the exact mechanism of ADE remains unclear.Methodology/Principal FindingVirus neutralization and ADE assays were performed using ultracentrifugation supernatants of acute-phase sera from patients with secondary infections or human monoclonal antibodies (HuMAbs) as anti-DENV antibodies. Virus sources included infectious serum-derived viruses from the ultracentrifugation precipitates, laboratory-culture adapted DENV, or recombinant DENVs derived from patient sera. In contrast to the high levels of ADE observed with laboratory virus strains, low ADE was observed with autologous patient-derived viruses, when patient sera were used to provide the antibody component in the ADE assays. Similar results were obtained using samples from DF and DHF patients. Recombinant-viruses derived from DHF patients showed only minor differences in neutralization and ADE activity in the presence of HuMAbs or plasma derived from the same DHF patient.Conclusion/SignificanceSerum or plasma taken from patients during the acute phase of a secondary infection showed high levels of ADE, but no neutralization activity, when assayed in the presence of laboratory-adapted virus strains. By contrast, serum or plasma from the same patient showed high levels of neutralization activity but failed to induce significant ADE when the assays were performed with autologous virus. These results demonstrate the significance of the virus source when measuring ADE. They also suggest that repeated passage of DENV in cell culture has endowed it with the capacity to induce high levels of ADE.

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Section: Discussionsupporting

confidence: 86%

“…Previous studies assayed ADE in K562 [10], [26], [34], THP-1 [23], [35]–[37] or BHK [38], [39] cells. Therefore, we used both THP-1 and K562 cells to assay ADE (Supplemental Figure S5).…”

Section: Resultsmentioning

confidence: 99%

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Low Levels of Antibody-Dependent Enhancement in Vitro Using Viruses and Plasma from Dengue Patients

et al. 2014

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Chikungunya Virus Growth and Fluorescent Labeling: Detection of Chikungunya Virus by Immunofluorescence Assay

Moi

Takasaki

2016

Methods in Molecular Biology

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Immunofluorescence assay (IFA) is a highly versatile and sensitive assay for detection and titration of chikungunya virus (CHIKV). The IFA technique requires virus-infected cells (viral antigen) and antibodies specific to the viral antigens for detection. Suitable antibodies for detection include monoclonal antibodies specific to CHIKV structural and nonstructural proteins, polyclonal antibodies, and convalescent serum samples. Here, the details of virus antigen preparation, detection by IFA method, and applications are described. The described IFA method is potentially useful in a wide range of studies including virus growth kinetics and virus infection mechanism studies. Additionally, the described IFA method can be modified for applications in arbovirus diagnosis, including CHIKV.

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Evaluation and analysis of dengue virus enhancing and neutralizing activities using simple high-throughput assays

Jing

Huang

et al. 2013

Appl Microbiol Biotechnol

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The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose-response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.

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Dengue virus isolation relying on antibody-dependent enhancement mechanism using FcγR-expressing BHK cells and a monoclonal antibody with infection-enhancing capacity

Cited by 6 publications

References 18 publications

Low Levels of Antibody-Dependent Enhancement in Vitro Using Viruses and Plasma from Dengue Patients

Low Levels of Antibody-Dependent Enhancement in Vitro Using Viruses and Plasma from Dengue Patients

Chikungunya Virus Growth and Fluorescent Labeling: Detection of Chikungunya Virus by Immunofluorescence Assay

Evaluation and analysis of dengue virus enhancing and neutralizing activities using simple high-throughput assays

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