Abstract
Human dendritic cells (DCs) are the main target cells of dengue virus (DENV). Because humans injected with even a small volume of DENV from mosquito saliva display a high level of viremia, DCs are expected to be highly susceptible to DENV. In the present study, we assessed the efficiency of DENV infection using the novel immortalized human myeloid cell lines iPS-ML and iPS-DC. To prepare the DC-like myeloid cell line (iPS-DC), iPS-ML cells were cultured in the presence of IL-4 for 72 h. iPS-DC cells were the most susceptible to DENV, followed by iPS-ML, Vero and K562 cells. In contrast, the highest infective yield titer was observed in Vero cells. To investigate further uses of iPS-ML and iPS-DC, these cells were applied to an assay measuring antibody-dependent enhancement (ADE) activity in DENV infection. Serum samples collected from healthy Thai participants and mouse monoclonal antibodies displayed similar ADE activity patterns when examined with iPS-ML, iPS-DC, or K562 cells, the last of which are usually used in conventional ADE assays. Interestingly, iPS-ML cells showed greater susceptibility to ADE activity than iPS-DC and K562 cells. Here, we demonstrated the potential utility of the novel immortalized human myeloid cell lines iPS-ML and iPS-DC in future research on DENV.
Highlights
Dengue virus (DENV) has four genetically distinct serotypes (DENV-1, DENV-2, DENV-3 and DENV-4), which belong to the genus Flavivirus, family Flaviviridae [1]
To investigate the susceptibility of induced pluripotent stem (iPS)-ML and iPS-dendritic cells (DCs) to DENVs, these cells were inoculated with DENV-1–4 at a multiplicity of infection (MOI) of 0.001, or mock infected (MOCK)
The virus titers in the supernatant collected from infectedcells at an MOI of 0.001 were shown in Figure 1B, suggesting that the yield levels obtained with DENV-3 and DENV-4 were higher than those with DENV-1 and DENV-2
Summary
Dengue virus (DENV) has four genetically distinct serotypes (DENV-1, DENV-2, DENV-3 and DENV-4), which belong to the genus Flavivirus, family Flaviviridae [1]. Before the dengue vaccine can be confidently administered to large populations worldwide, there is need of an appropriate antibody survey system to accurately and sensitively determine the seronegative/seropositive status, and the functional antibody status (protective neutralization or pathogenic ADE) of potential vaccinees. DENV, which is transmitted by infected mosquitoes, initially targets dendritic cells (DCs) in human skin [8, 9]. As the initial target cells of DENV, DCs are critical for primary replication in the human epidermis/dermis during the early infection phase [14, 15]. Another group has demonstrated that primary DCs showed ADE in DENV infection [16]. We assessed two novel immortalized human myeloid cell lines to determine whether they have susceptibility to DENV and could be useful in an antibody assay to detect ADE activity
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