Leishmania donovani activates nuclear transcription factor-kappaB in macrophages through reactive oxygen...

Interaction of Leishmania donovani with macrophages antagonizes host defense mechanisms by interfering with a cascade of cell signaling processes in the macrophages. An early intracellular signaling event that follows receptor engagement is the activation of transcription factor NF-kappaB. It has been reported earlier that NF-kappaB-dependent signaling pathway regulates proinflammatory cytokine release. We therefore investigated the effect of L. donovani infectivity on this nuclear transcription factor in macrophage cell line J774A.1. Both L. donovani and its surface molecule lipophosphoglycan (LPG) resulted in a dose- and time-dependent activation of NF-kappaB-DNA binding activity in an electrophoretic mobility shift assay. We also report the involvement of IkappaB-alpha and IkappaB-beta in the persistent activation of NF-kappaB by L. donovani. We demonstrate that the NF-kappaB activation was independent of viability of the parasite. Electrophoretic mobility supershift assay indicated that the NF-kappaB complex consists of p65 and c-rel subunits. The interaction of parasite with the macrophages and not the cellular uptake was important for NF-kappaB activation. Both p38 and ERK mitogen activated protein kinase (MAP) activation appears to be necessary for NF-kappaB activation by LPG. Preincubation of cells with antioxidants resulted in inhibition of L. donovani induced NF-kappaB activation, thereby suggesting a potential role of reactive oxygen species in L. donovani induced intracellular signaling. The present data indicate that antioxidants could play an important role in working out various therapeutic modalities to control leishmaniasis.