Analysis of the Response of Human Neonatal Dendritics Cells and Macrophages Against Rotavirus (137.26)

Abstract

Rotavirus (RV) is the most common cause of infectious diarrhea. Dendritic cells (DCs) and macrophages play a crucial role in the immune response against viral infections; therefore it is important to dissect the interaction of RV with these cells. Here we analyzed the interaction of RV with neonatal DCs and macrophages in an in vitro model. Cord blood-derived mononuclear cells were culture in vitro for 6 days in the presence of either hGM-CSF/IL-4 or hM-CSF to generated enriched DCs and macrophages, respectively. These cells were cultured for 24 hrs with rotavirus and the levels of the activation markers CD40, CD86 and MHC class-II, and the intracellular cytokines IL-6, IL-12 and IL-10 were analyzed by flow cytometry. TGF-β was evaluated by ELISA. Finally, the induction of allogenic T-celll proliferation was analyzed. We found that RV could induce the activation of the neonatal DCs and macrophages since all the activation markers analyzed were up regulated, albeit at different levels. As result of this activation, the neonatal DCs and macrophages produced high levels of the regulatory cytokines, but moderate levels of pro-inflammatory cytokines. Finally, the activated DCs and macrophages were able to induce a strong allogenic T-cell proliferation. These results showed that neonatal DCs and macrophages could be activated by RV and that this activation was sufficient to induce the priming of allogenic T-cells. Supported by grant SEP-CONACYT (80149) and DGAPA, UNAM.