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Piera, K.

Publications and source records attributed to Piera, K..

2 recordsLinked to original sources

Age is an intrinsic driver of inflammatory responses to malaria

Age is a critical factor influencing the host immune response to infection and disease pathogenesis. In malaria, the risk of severe disease increases with age in non-immune individuals. Malaria severity is in part driven by inflammation, but the specific cells and mechanisms contributing to age-dependent disease risk are incompletely understood. Here, we assessed inflammatory cytokines in non-immune children and adults with clinical malaria, and the phenotypic, functional and transcriptional differences of in vitro innate cell responders to malaria parasites in naive children and adults. During naturally acquired malaria, age was associated with increased plasma levels of inflammatory chemokines CCL2, CCL3, CXCL8, CXLC9, along with CRP, and IDO, which were associated with clinical symptoms. In malaria naive individuals, classical monocyte and V{delta}2+ {gamma}{delta} T cell responses from adults were characterized by higher inflammatory cytokine production, and transcriptional activation following stimulation with malaria parasites. Classical monocyte responses in adults were dominated by CCL2 production, while in children the response had increased IL10 production and enrichment in IL10 signaling pathways upon parasite stimulation. This heightened inflammatory response in adults was not mitigated by parasite induced Tregs. Taken together, these findings identify cellular mechanisms of age-dependent host responses that play crucial roles in driving inflammatory responses in malaria.

immunology↗

Implementation of a novel optogenetic tool in mammalian cells based on a split T7 RNA polymerase

Optogenetic tools are widely used to control gene expression dynamics both in prokaryotic and eukaryotic cells. These tools are used in a variety of biological applications from stem cell differentiation to metabolic engineering. Despite some tools already available in bacteria, no light-inducible system currently exists to orthogonally control gene expression in mammalian cells. Such a tool would be particularly important in synthetic biology, where orthogonality is advantageous to achieve robust activation of synthetic networks. Here we implement, characterize and optimize a new orthogonal optogenetic tool in mammalian cells based on a previously published system in bacteria called Opto-T7RNAPs. The tool consists of a split T7 RNA polymerase coupled with the blue light-inducible magnets system (mammalian OptoT7 - mOptoT7). In our study we exploited the T7 polymerases viral origins to tune our systems expression level, reaching up to 20-fold change activation over the dark control. mOptoT7 is used here to generate mRNA for protein expression, shRNA for protein inhibition and Pepper aptamer for RNA visualization. Moreover, we show that mOptoT7 can mitigate gene expression burden when compared to other optogenetic constructs. These properties make mOptoT7 a new powerful tool to use when orthogonality and viral-like RNA species are desired in both synthetic biology and basic science applications.

synthetic biology↗