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Scheeres, E. C.

Publications and source records attributed to Scheeres, E. C..

2 recordsLinked to original sources

TGFβ primes alveolar-like macrophages to induce type I IFN following TLR2 activation

Alveolar macrophages (AMs) are key mediators of lung function and are potential targets for therapies during respiratory infections. TGF{beta} is an important regulator of AM differentiation and maintenance, but how TGF{beta} directly modulates the innate immune responses of AMs remains unclear. This shortcoming prevents effective targeting of AMs to improve lung function in health and disease. Here we leveraged an optimized ex vivo AM model system, fetal-liver derived alveolar-like macrophages (FLAMs), to dissect the role of TGF{beta} in AMs. Using transcriptional analysis, we first globally defined how TGF{beta} regulates gene expression of resting FLAMs. We found that TGF{beta} maintains the baseline metabolic state of AMs by driving lipid metabolism through oxidative phosphorylation and restricting inflammation. To better understand inflammatory regulation in FLAMs, we next directly tested how TGF{beta} alters the response to TLR2 agonists. While both TGF{beta} (+) and TGF{beta} (-) FLAMs robustly responded to TLR2 agonists, we found an unexpected activation of type I interferon (IFN) responses in FLAMs and primary AMs in a TGF{beta}-dependent manner. Surprisingly, mitochondrial antiviral signaling protein and the interferon regulator factors 3 and 7 were required for IFN production by TLR2 agonists and the IFN response was dependent on mitochondrial reactive oxygen species. Together, these data suggest that TGF{beta} modulates AM metabolic networks and innate immune signaling cascades to control inflammatory pathways in AMs.

immunology↗

The Trichloroethylene Metabolite S-(1,2-dichlorovinyl)-l-cysteine Inhibits Lipopolysaccharide-induced Transcriptomic Inflammation Pathways and Cytokine Secretion in a Macrophage Cell Model

Previous studies have shown that the trichloroethylene metabolite S-(1,2-dichlorovinyl)-l-cysteine (DCVC) inhibits cytokine secretion in pathogen stimulated fetal membrane tissue but little is known about the mechanism for these effects, including which cell types or transcriptomic pathways are impacted. Macrophages play a critical role in the fetal membrane innate immune response during infection. We tested the hypothesis that DCVC inhibits lipopolysaccharide (LPS) stimulated inflammation pathways in differentiated (macrophage-like) THP-1 cells. THP-1 cells were differentiated with phorbol 12-myristate 13-acetone for 24 hours and subsequently treated with 1, 5, or 10 {micro}M DCVC for 24 hours. After an additional 4 hour incubation with lipopolysaccharide (LPS), we collected RNA and cell media. We performed transcriptomic analysis using RNA sequencing analysis for 5{micro}M DCVC treatments and quantified cytokine release (IL-1{beta}, IL-6, and TNF-) into cell media for 1, 5 and 10 {micro}M DCVC treatments. RNAseq analysis revealed 1,399 differentially expressed genes (FDR<0.05 and log2fold change magnitude>2.5) in the cells co-treated with DCVC and LPS compared to LPS alone. For example, TNF was 9-fold downregulated with the addition of DCVC. Major pathways downregulated (adjusted p-value<0.05) in DCVC+LPS treatments versus LPS-only treatments, included: "acute inflammatory response", "production of molecular mediator of immune response" and "phagocytosis". LPS increased IL-1{beta}, IL-6, and TNF- levels in culture media (p<0.001), but this effect which was inhibited by co-treatment with DCVC (p<0.001 for LPS vs. LPS+DCVC treatments). Our results demonstrate that DCVC suppresses inflammatory responses in macrophages.

pharmacology and toxicology↗