ABSTRACT
BACKGROUND AND PURPOSE: Cell-to-cell interactions between mast cells and activated T cells are increasingly recognized as a possible mechanism in the aetiology of allergic or non-allergic inflammatory disorders. To determine the anti-allergic effect of fisetin, we examined the ability of fisetin to suppress activation of the human mast cell line, HMC-1, induced by activated Jurkat T cell membranes. EXPERIMENTAL APPROACH: HMC-1 cells were incubated with or without fisetin for 15 min and then co-cultured with Jurkat T cell membranes activated by phorbol-12-myristate 13-acetate for 16 h. We determined gene expression in activated HMC-1 cells by DNA microarray and quantitative reverse transcription (RT)-PCR analysis. We also examined activation of the transcription factor NF-kappaB and MAP kinases (MAPKs) in activated HMC-1 cells. KEY RESULTS: Fisetin suppresses cell spreading and gene expression in HMC-1 cells stimulated by activated T cell membranes. Additionally, we show that these stimulated HMC-1 cells expressed granzyme B. The stimulatory interaction also induced activation of NF-kappaB and MAPKs; these activations were suppressed by fisetin. Fisetin also reduced the amount of cell surface antigen CD40 and intercellular adhesion molecule-1 (ICAM-1) on activated HMC-1 cells. CONCLUSIONS AND IMPLICATIONS: Fisetin suppressed activation of HMC-1 cells by activated T cell membranes by interfering with cell-to-cell interaction and inhibiting the activity of NF-kappaB and MAPKs and thereby suppressing gene expression. Fisetin may protect against the progression of inflammatory diseases by limiting interactions between mast cells and activated T cells.
Subject(s)
Cell Membrane/physiology , Flavonoids/pharmacology , Mast Cells/drug effects , T-Lymphocytes/physiology , CD40 Antigens/biosynthesis , Cell Degranulation , Cell Line , Cell Line, Tumor , Flavonols , Granzymes/biosynthesis , Humans , I-kappa B Kinase/metabolism , Intercellular Adhesion Molecule-1/biosynthesis , Mast Cells/cytology , Mast Cells/physiology , Mitogen-Activated Protein Kinases/metabolism , NF-kappa B/metabolism , Oligonucleotide Array Sequence Analysis , Perforin/biosynthesis , Phosphorylation , T-Lymphocytes/ultrastructureSubject(s)
Crohn Disease/complications , Urinary Bladder, Neurogenic/etiology , Adult , Humans , MaleABSTRACT
To examine the expression of heat shock protein (hsp) in each organ on autopsy in 11 deaths from hypothermia, we performed immunological staining of specimens from each organ using an antibody to hsp, ubiquitin (Ub). Staining of the liver, kidneys, lungs and pancreas revealed a high level of Ub expression in the cytoplasm and nuclei. However, staining of the same specimens of all controls revealed no Ub expression in the cytoplasm or nuclei. This finding suggests that cells are affected by stress even at a low temperature, and may be important in clarifying the cellular kinetics in death from hypothermia.
