UNLABELLED: The integrity of lipid rafts in cell membranes is important for signal transduction. AIM: To determine the distinct effects of beta-glycolipids on the composition of lipid rafts in natural killer T (NKT) cells and on the level of expression of flotillin-2, leukocyte-specific protein tyrosine kinase (Lck), and STAT1-associated pathways. METHODS: The effects of glycolipids were determined by composition analysis of the raft domains, FACS analysis of the distribution patterns for the raft ganglioside, GM1, and fluorescence microscopy of raft patching. To evaluate the effects of the immune environment on glycolipid-associated alteration of lipid rafts, hepatitis was induced by an intravenous injection of concanavalin A (ConA) in mice treated with various glycolipids. RESULTS: The administration of beta-glucosylceramide, beta-lactosylceramide, and a combination of both significantly altered GM1 content in lymphocyte membranes in an environment-dependent manner. These effects were associated with altered expression levels of flotillin-2, Lck, and STAT1, and with a significant decrease in intrahepatic CD8+ lymphocyte trapping and the alleviation of ConA-induced hepatitis. The administration of alpha-glycolipids failed to induce similar effects. CONCLUSIONS: The alteration in the expression levels of flotillin-2, Lck, and STAT1 that occurs concomitantly with changes in lipid raft composition and structure following the administration of beta-glycolipids in ConA-induced hepatitis is microenvironment-dependent and is associated with decreased intrahepatic CD8(+) T lymphocyte trapping and amelioration of immune-mediated hepatitis.
UNLABELLED: The integrity of lipid rafts in cell membranes is important for signal transduction. AIM: To determine the distinct effects of beta-glycolipids on the composition of lipid rafts in natural killer T (NKT) cells and on the level of expression of flotillin-2, leukocyte-specific protein tyrosine kinase (Lck), and STAT1-associated pathways. METHODS: The effects of glycolipids were determined by composition analysis of the raft domains, FACS analysis of the distribution patterns for the raft ganglioside, GM1, and fluorescence microscopy of raft patching. To evaluate the effects of the immune environment on glycolipid-associated alteration of lipid rafts, hepatitis was induced by an intravenous injection of concanavalin A (ConA) in mice treated with various glycolipids. RESULTS: The administration of beta-glucosylceramide, beta-lactosylceramide, and a combination of both significantly altered GM1 content in lymphocyte membranes in an environment-dependent manner. These effects were associated with altered expression levels of flotillin-2, Lck, and STAT1, and with a significant decrease in intrahepatic CD8+ lymphocyte trapping and the alleviation of ConA-induced hepatitis. The administration of alpha-glycolipids failed to induce similar effects. CONCLUSIONS: The alteration in the expression levels of flotillin-2, Lck, and STAT1 that occurs concomitantly with changes in lipid raft composition and structure following the administration of beta-glycolipids in ConA-induced hepatitis is microenvironment-dependent and is associated with decreased intrahepatic CD8(+) T lymphocyte trapping and amelioration of immune-mediated hepatitis.
Authors: Meir Mizrahi; Tomer Adar; Gadi Lalazar; Dean Nachman; Madi El Haj; Ami Ben Ya'acov; Yoav Lichtenstein; Yehudit Shabat; Dimitri Kanovich; Lida Zolotarov; Yaron Ilan Journal: J Clin Transl Hepatol Date: 2018-02-14