xCT-mediated glutamate excretion in white adipocytes stimulates interferon-γ production by natural killer cells in obesity.

Obesity-mediated hypoxic stress underlies inflammation, including interferon (IFN)-γ production by natural killer (NK) cells in white adipose tissue. However, the effects of obesity on NK cell IFN-γ production remain obscure. Here, we show that hypoxia promotes xCT-mediated glutamate excretion and C-X-C motif chemokine ligand 12 (CXCL12) expression in white adipocytes, resulting in CXCR4+ NK cell recruitment. Interestingly, this spatial proximity between adipocytes and NK cells induces IFN-γ production in NK cells by stimulating metabotropic glutamate receptor 5 (mGluR5). IFN-γ then triggers inflammatory activation of macrophages and augments xCT and CXCL12 expression in adipocytes, forming a bidirectional pathway. Genetic or pharmacological inhibition of xCT, mGluR5, or IFN-γ receptor in adipocytes or NK cells alleviates obesity-related metabolic disorders in mice. Consistently, patients with obesity showed elevated levels of glutamate/mGluR5 and CXCL12/CXCR4 axes, suggesting that a bidirectional pathway between adipocytes and NK cells could be a viable therapeutic target in obesity-related metabolic disorders.

Isolation of Hepatic Stellate Cells and Lymphocytes for Co-culture Systems.

In contrast to quiescent hepatic stellate cells (HSCs), activated HSCs play crucial roles in the development of liver fibrosis by producing a huge amount of extracellular matrix such as collagen fibers. However, recent lines of evidence have also highlighted the immunoregulatory functions of HSCs, in which they interact with diverse hepatic lymphocytes to produce cytokines and chemokines, release extracellular vesicles, or express specific ligands. Therefore, to understand the exact interactions between HSCs and lymphocyte subsets in the pathogenesis of the liver disease, it is valuable to establish experimental procedures to isolate HSC and co-culture them with lymphocytes. Here, we introduce the efficient methods to isolate and purify mouse HSCs and hepatic lymphocytes using density gradient centrifugation, microscopic observation, and flow cytometry. Moreover, we provide the direct and indirect co-culturing methods of isolated mouse HSCs and hepatic lymphocytes based upon the purpose of the study.

Catecholamine induces Kupffer cell apoptosis via growth differentiation factor 15 in alcohol-associated liver disease.

Chronic alcohol consumption often induces hepatic steatosis but rarely causes severe inflammation in Kupffer cells (KCs) despite the increased hepatic influx of lipopolysaccharide (LPS), suggesting the presence of a veiled tolerance mechanism. In addition to LPS, the liver is affected by several gut-derived neurotransmitters through the portal blood, but the effects of catecholamines on KCs have not been clearly explored in alcohol-associated liver disease (ALD). Hence, we investigated the regulatory roles of catecholamine on inflammatory KCs under chronic alcohol exposure. We discovered that catecholamine levels were significantly elevated in the cecum, portal blood, and liver tissues of chronic ethanol-fed mice. Increased catecholamines induced mitochondrial translocation of cytochrome P450 2E1 in perivenous hepatocytes expressing the ß2-adrenergic receptor (ADRB2), leading to the enhanced production of growth differentiation factor 15 (GDF15). Subsequently, GDF15 profoundly increased ADRB2 expression in adjacent inflammatory KCs to facilitate catecholamine/ADRB2-mediated apoptosis. Single-cell RNA sequencing of KCs confirmed the elevated expression of Adrb2 and apoptotic genes after chronic ethanol intake. Genetic ablation of Adrb2 or hepatic Gdf15 robustly decreased the number of apoptotic KCs near perivenous areas, exacerbating alcohol-associated inflammation. Consistently, we found that blood and stool catecholamine levels and perivenous GDF15 expression were increased in patients with early-stage ALD along with an increase in apoptotic KCs. Our findings reveal a novel protective mechanism against ALD, in which the catecholamine/GDF15 axis plays a critical role in KC apoptosis, and identify a unique neuro-metabo-immune axis between the gut and liver that elicits hepatoprotection against alcohol-mediated pathogenic challenges.

Dependency of Tunneling-Magnetoresistance Ratio on Nanoscale Spacer Thickness and Material for Double MgO Based Perpendicular-Magnetic-Tunneling-Junction.

It was found that in double MgO based perpendicular magnetic tunneling junction spin-valves ex-situ annealed at 400 °C, the tunneling magnetoresistance ratio was extremely sensitive to the material and thickness of the nanoscale spacer: it peaked at a specific thickness (0.40~0.53 nm), and the TMR ratio for W spacers (~134%) was higher than that for Ta spacers (~98%). This dependency on the spacer material and thickness was associated with the (100) body-centered-cubic crystallinity of the MgO layers: the strain enhanced diffusion length in the MgO layers of W atoms (~1.40 nm) was much shorter than that of Ta atoms (~2.85 nm) and the shorter diffusion length led to the MgO layers having better (100) body-centered-cubic crystallinity.