This nding could have implications for the susceptibility of DENV contaminated individuals to other blood pathogens, this kind of as HIV or hepatitis C virus , considering that a lack of IFN manufacturing in those cells may possibly let a secondary infection to progress additional efciently. The obser vation that type I IFN gene expression is diminished in DENV infected DCs and in addition in 293T cells following triggering of IFN production clearly indicates that DENV infection interferes with this particular pathway. Also supporting this observation certainly is the requirement of DENV replication for this inhibition, since UV inactivation of DENV completely abol ishes this inhibition.
Due to the fact this inhibition was ob served soon after IFN induction by various pathways , such as RIG I, MDA5, and TLR3, it could indicate that selelck kinase inhibitor DENV interferes with these pathways by focusing on a typical component or that DENV may possibly encode extra IFN antag onists focusing on each one of those pathways at different ranges, as is described for that WNV E, NS1, and NS2A pro teins. On the whole, virally encoded proteins may well have a few functions, and the same viral immune antagonist can interfere with a few pathways. For example, the inuenza A NS1 protein has various functions, this kind of as the inhibition within the style I IFN program in contaminated cells, binding and sequestra tion of dsRNA,

interference with host mRNA processing, fa cilitation of preferential viral mRNA translation, and inhibi tion of DC activation. Our data showing DENV interference with IFN produc tion in contaminated DCs even at the IFN RNA degree recommend that DENV infection interferes with the kind I IFN manufacturing pathway at an upstream step just before the induction of gene expression and not on the protein degree.
The reduction of IRF 3 phosphorylation observed just after NDV infection in previously DENV infected DCs when compared to outcomes for only NDV infected ones or in DENV infected 293T cells after SeV infection supports this hypothesis. Given that selleck inhibitor some DCs exposed to DENV had been not infected with DENV but had been subsequently contaminated with NDV , that is in a position to in duce a strong IRF three phosphorylation , it may be difcult to demonstrate a strong reduction of phosphorylated IRF three after NDV infection of previously DENV infected DCs by Western blotting. Also, it could be difcult to distinguish the contribution to your IRF three phosphorylation of each population present in the group of doubly infected DCs.
Really, the 37% reduction observed just after quantication of the Western blot densitometry ought to corre spond for the number of DCs which have been coinfected using the two viruses. If we assume that equal levels of NDV infection induce very similar ranges of IRF 3 phosphorylation, the main difference observed in IRF three phosphorylation in between NDV infected DCs and DENV NDV infected DCs cannot be due to a distinction in NDV infection ranges, because the percentages of NDV infected cells are very similar.