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  • Title: Enhanced TNF alpha production by monocytic-like cells exposed to dengue virus antigens.
    Author: Hober D, Shen L, Benyoucef S, De Groote D, Deubel V, Wattré P.
    Journal: Immunol Lett; 1996 Nov; 53(2-3):115-20. PubMed ID: 9024988.
    Abstract:
    The studies indicating the importance of TNF alpha in dengue virus infection have led us to determine whether monocyte-like cells produce TNF alpha exposure after dengue virus. The supernatant fluids of mosquito cells (AP61) infected with dengue virus (DV) type 1 and DV type 3 were harvested 7 days post-infection and clarified. DV inactivation was performed in the presence of betapropiolactone that preserves antigenicity of viruses. We used the monocytic-like cell line THP-1 that is a model system of TNF alpha production. Polymyxin B (50 micrograms/ml) was added to block untoward effects resulting from possible LPS contamination of media or cultures. THP-1 cells were primed with a phorbol ester (PMA) for 24 h, then they were cultured for 4 and 24 h in the presence of inactivated culture supernatant of dengue infected AP61 cells or control preparations. The concentrations of TNF alpha in the culture supernatants were measured by using an immunoenzymatic assay. PMA-treated THP-1 cells rapidly secreted TNF alpha in response to inactivated culture supernatant of DV-infected cells. We found high levels of TNF alpha with cells exposed to DV1 and DV3 preparations compared with controls (mean values; 465 and 829 vs. 70 pg/ml, respectively, at 24 h post exposure, n = 4). We obtained a substantial inhibition of the enhancing activity of DV1 and DV3 infected supernatants in the presence of dengue hyperimmune mouse ascitic fluids. Our results demonstrate that exposure of monocytes/macrophages to DV particles or virus proteins derived from DV may be responsible for the enhanced production of TNF alpha in DV-infected patients.
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