A model of the real-time correlation of viral titers with immune reactions in antibody-dependent enhancement of dengue-2 infections

Abstract

We simultaneously assessed dengue-2 virus (DEN-2) titers by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immune reactions including interleukin-4 (IL-4), interferon-gamma (IFN-gamma) and prostaglandin E(2) (PGE(2)) production by human mononuclear cells (MNLs) in a model of antibody-dependent enhancement (ADE). We found that DEN-1 immune sera at 1:100 and 1:250, but not those at 1:10 or control sera, enhanced DEN-2 infections in human MNLs as assessed by the fluorogenic RT-PCR technique. The enhanced profiles of DEN-2 infections determined by the RT-PCR in 6 h were reproducible by the standard plaque-forming unit (PFU) measurement established after 7 days. The ADE-enhanced DEN-2 titers determined by the RT-PCR were 5.5-33-fold higher than those detected by PFU assay, suggesting that total virions during infections were much higher than the viable ones detected by PFU assay. MNLs in response to DEN-2 infections had higher IFN-gamma and PGE(2) production. However, the enhancement of DEN-2 infections by DEN-1 immune sera in MNLs was not associated with further enhancement of IFN-gamma production. In contrast, the presence of subneutralizing DEN-1 immune sera that enhanced DEN-2 infections also enhanced PGE(2) but not IL-4 production. The results of this study suggest that ADE of DEN-2 infections associated with induction of immunosuppressive mediators such as PGE(2) and IL-4 can be simultaneously assessed in a real-time fashion.