The influence of chronic alcohol consumption on invariant natural killer T cells

Chronic alcohol consumption increases the incidence of infectious diseases and multiple types of cancer and decreases the survival of cancer patients, which is associated with a compromised host immune system. Invariant natural killer T (iNKT) cells are an important population of immunoregulatory cells and play important roles in shaping immune response in infectious diseases and cancers. However, the knowledge of alcohol’s effects on iNKT cells is very limited. Using a well-established experimental mouse model, we found that alcohol consumption in the steady state increases iNKT cells in the thymus and liver, enhances iNKT cell proliferation, activation and maturation. Upon activation, iNKT cells exhibit a Th1-dominant cytokine profile, and enhance Th1 response evidenced by the enhanced activation of dendritic cells, NK, T, and B cells, which favors anti-tumor immunity. In addition, alcohol consumption increases the expression of chemokine receptor CXCR3 on iNKT cells in the thymus, which could be associated with increased thymic retention of iNKT cells. Moreover, alcohol consumption increases inhibitory receptor NKG2A-expressing iNKT cells, which could be a negative feedback mechanism to prevent iNKT cell over-activation. In contrast to the steady state, with tumor progression alcohol consumption accelerates iNKT cell anergy and switches iNKT cell cytokine profile from Th1-dominant to Th2-dominant, which favors tumor progression and decreases the survival of melanoma-bearing mice. In subsequent mechanistic studies, we show that alcohol consumption increases C16-alkenyl-LPE, an endogenous lipid antigen important for iNKT cell development, in the thymus. This finding supports our hypothesis that alcohol consumption influences iNKT cells by altering their endogenous lipid antigens in the thymus. We further show that TNF-α, but not IFN-γ, mediates the alcohol-induced increase of NKG2A+ iNKT in the thymus and spleen and the alcohol-induced increase of CXCR3+ iNKT in the thymus. In summary, chronic alcohol consumption enhances iNKT cell proliferation, activation and maturation via increasing endogenous lipid antigens, which not only enhances iNKT cell Th1 response and favors antitumor immunity, it also predisposes iNKT cells to accelerated immunosenescence. Crosstalk with melanoma, alcohol accelerates iNKT cells anergy and favors tumor progression. TNF-a may play the key role in alcohol-induced increase in CXCR3+ and NKG2A+ iNKT cells

Chronic alcohol consumption inhibits peripheral NK cell development and maturation by decreasing the availability of IL-15

NK cells are innate immune cells and have important roles in antiviral and antitumor immunity. Based on the transcriptional regulation, organ distribution, and cell function, NK cells have recently been further divided into cytotoxic conventional NK cells (cNK) and noncytotoxic helper-like group 1 innate lymphoid cells (ILC1s). It is well known that chronic alcohol consumption decreases peripheral NK cell number and cytolytic activity; however, the underlying mechanism remains to be elucidated. How chronic alcohol consumption affects ILC1s is, to our knowledge, completely unexplored. Herein, we used a well-established mouse model of chronic alcohol consumption to study the effects of alcohol on transcription factor expression, maturation, and cytokine production of cNK cells and ILC1s in various organs. We found that alcohol consumption significantly decreased Eomes-expressing cNK cells in all the examined organs, except BM, but did not significantly affect ILC1s. Alcohol consumption compromised cNK cell development and maturation. Exogenous IL-15/IL-15Rα treatment caused full recovery of Eomes-expressing cNK cell number and maturation. Taken together, our data indicated that chronic alcohol consumption decreases cNK cell number and cytolytic activity by arresting cNK cell development at the CD27 CD11b stage. This developmental arrest of NK cells results from a lack of IL-15 availability in the microenvironment. IL-15/IL-15Rα treatment can recover alcohol consumption-induced developmental defect in NK cells

Isotope dilution mass spectrometry for the quantification of sulfane sulfurs

Sulfane sulfurs are one type of important reactive sulfur species. These molecules have unique reactivity that allows them to attach reversibly to other sulfur atoms and exhibit regulatory effects in diverse biological systems. Recent studies have suggested that sulfane sulfurs are involved in signal transduction processes regulated by hydrogen sulfide (H2S). Accurate and reliable measurements of sulfane sulfurs in biological samples are thus needed to reveal their production and mechanisms of actions. Herein we report a convenient and accurate method for the determination of sulfane sulfur concentrations. The method employs a triphenylphosphine derivative (P2) to capture sulfane sulfurs as a stable phosphine sulfide product, PS2. The concentration of PS2 was then determined by isotope dilution mass spectrometry, using a (13)C3-labeled phosphine sulfide, PS1, as the internal standard. The specificity and efficiency of the method were proven by model reactions. It was also applied to the measurement of sulfane sulfurs in mouse tissues including brain, kidney, lung, liver, heart, spleen, and blood

Integrated Multifunctional Laryngoscope for Medical Diagnosis and Treatment

Laryngeal lesions can cause great inconvenience to patients. Early diagnosis and corresponding treatments are critical to the survival of patients. However, the diagnosis and precise removal of tumors remain a challenge under the use of a white light laryngoscope. In this work, an integrated, multifunctional laryngoscope was designed and tested for the imaging evaluation and precision laser surgery for laryngeal tissue. This integrated diagnostic and therapeutic endoscopic system included two imaging modes (i.e., optical coherence tomography and white light endoscopy) and a laser ablation treatment mode. The endoscope had a common-path design to ensure that the same position could be imaged and treated simultaneously. The ex vivo porcine larynx experimental results showed that the system imaging modes could simultaneously acquire both superficial and cross-sectional images of the sample tissue, and the ablation treatment could be performed under imaging guidance. This multifunctional laryngoscope has great potential for the early diagnosis of and accurate laser ablation surgery for laryngeal tumors

Radiation-Induced Innate Neutrophil Response in Tumor Is Mediated by the CXCLs/CXCR2 Axis

The early events that lead to the inflammatory and immune-modulatory effects of radiation therapy (RT) in the tumor microenvironment (TME) after its DNA damage response activating the innate DNA-sensing pathways are largely unknown. Neutrophilic infiltration into the TME in response to RT is an early innate inflammatory response that occurs within 24–48 h. Using two different syngeneic murine tumor models (RM-9 and MC-38), we demonstrated that CXCR2 blockade significantly reduced RT-induced neutrophilic infiltration. CXCR2 blockade showed the same effects on RT-induced tumor inhibition and host survival as direct neutrophil depletion. Neutrophils highly and preferentially expressed CXCR2 compared to other immune cells. Importantly, RT induced both gene and protein expression of CXCLs in the TME within 24 h, attracting neutrophils into the tumor. Expectedly, RT also upregulated the gene expression of both cGAS and AIM2 DNA-sensing pathways in cGAS-positive MC-38 tumors but not in cGAS-negative RM-9 tumors. Activation of these pathways resulted in increased IL-1β, which is known to activate the CXCLs/CXCR2 axis. Gene ontology analysis of mRNA-Seq supported these findings. Taken together, the findings suggest that the CXCLs/CXCR2 axis mediates the RT-induced innate inflammatory response in the TME, likely translating the effects of innate DNA-sensing pathways that are activated in response to RT-induced DNA damage

FoxO3a inhibiting expression of EPS8 to prevent progression of NSCLC: A new negative loop of EGFR signalingResearch in context

Background: The resistance to EGF receptor (EGFR) tyrosine kinase inhibitors (TKI) is a major challenge in the treatment of non-small cell lung cancer (NSCLC). Understanding the molecular mechanisms behind resistance is therefore an important issue. Here we assessed the role of EGFR pathway substrate 8 (EPS8) and Forkhead box O 3a (FoxO3a) as potentially valuable targets in the resistance of NSCLC . Methods: The expression levels of EPS8 and FoxO3a in patients with NSCLC (n = 75) were examined by immunohistochemistry staining, while in cells were detected by qPCR and western blot. The effects of EPS8 and FoxO3a on resistance, migration and invasion, cell cycle arrest were detected by MTT, transwell and flow cytometry, respectively. Chromatin immunoprecipitation and luciferase reporter assays were performed to determine the mechanisms of EPS8 expression and FoxO3a regulation. Findings: We observed that the expression of EPS8 inversely correlated with FoxO3a in NSCLC cell lines and NSCLC patients. FoxO3a levels were significantly decreased in tumor tissues compared with para-carcinoma tissues, while EPS8 is opposite. Besides, they play reverse roles in the resistance to gefitinib, the migration and invasion abilities, the cell cycle arrest in vitro and the tumor growth in vivo. Mechanistically, FoxO3a inhibits EPS8 levels by directly binding its gene promoter and they form a negative loop in EGFR pathway. Interpretation: Targeting FoxO3a and EPS8 in EGFR signaling pathway prevents the progression of NSCLC, which implied that the negative loop they formed could served as a therapeutic target for overcoming resistance in NSCLC. Funds: National Natural Science Foundation of China, Science and Technology Project of Henan, Outstanding Young Talent Research Fund of Zhengzhou University and the National Scholarship Fund. Keywords: Forkhead box O 3a (FoxO3a), Epidermal growth factor receptor (EGFR), EGFR pathway substrate 8 (EPS8), Gefitinib, Tumor resistance, Non-small cell lung cancer (NSCLC

Isotope dilution mass spectrometry for the quantification of sulfane sulfurs

Sulfane sulfurs are one type of important reactive sulfur species. These molecules have unique reactivity that can attach reversibly to other sulfur atoms and exhibit regulatory effects in diverse biological systems. Recent studies have suggested that sulfane sulfurs are involved in signal transduction processes regulated by hydrogen sulfide (H(2)S). Accurate and reliable measurements of sulfane sulfurs in biological samples are thus needed to reveal their production and mechanisms of actions. Herein we report a convenient and accurate method for the determination of sulfane sulfurs concentrations. The method employs a triphenylphosphine derivative (P2) to capture sulfane sulfurs as a stable phosphine sulphide product PS2. The concentration of PS2 was then determined by isotope dilution mass spectrometry, using a (13)C(3)-labelled phosphine sulfide PS1 as the internal standard. The specificity and efficiency of the method were proved by model reactions. It was also applied in the measurement of sulfane sulfurs in mice tissues including brain, kidney, lung, liver, heart, spleen, and blood