Role of MAPK signaling on human mast cell activation induced by Trichomonas vaginalis / Шинэ санаа Шинэ нээлт

Trichomonas vaginalis is a flagellated protozoan parasite that causes vaginitis and cervicitis in women and asymptomatic urethritis and prostatitis in men. Mast cells have been reported to be predominant in the vaginal smears and vaginal walls of patients infected with T. vaginalis. Mitogen activated protein kinase (MAPK) activated by various stimuli also regulate the transcriptional activity of various cytokine genes in the mast cells. Because of their essential role in intracellular signaling network, also appropriate targets for pharmacological treatment of inflammatory disorders.Cultivation of T.vaginalis and HMC-1 line, preparation of TvSP, to check intracellular ROS level and degranulation by FACS, to determine phosphorylation of MAPK and p47phox by immunobloting.In this study, we investigated whether MAPK were involved ROS generation and exocytotic degranulation in HMC-1 induced by T. vaginalis-derived secretory products (TvSP). We first examined that TvSP could induce activation of MAPK and NADPH oxidase in HMC-1 cells. Stimulation with TvSP induced phosphorylation of MAPK and p47phox in HMC-1 cells. Phosphorylation of p47phox is main source of ROS generation. Next, to determine involvement activation of MAPK in ROS generation and degranulation in HMC-1 cells induced by TvSP. ROS generation is required for exocytotic degranulation of mast cells induced by TvSP. Stimulation with TvSP induced phosphorylation of p47phox, ROSgeneration, and surface up-regulation of CD63 in human mast cells. CD63 is a marker for exocytosis. Pretreatment with MAPK inhibitors strongly inhibited TvSP-induced ROS generation and exocytotic degranulation.Our results suggest that TvSP could induce intracellular ROS generation and exocytotic degranulation in HMC-1 via MAPK signaling pathway.

The Influence of IgE on Cultured Human Mast Cells

PURPOSE: The mast cell plays a pivotal role in the human immune response. Crosslinking of 2 IgE molecules bound to the high affinity IgE receptor (FcepsilonRI) on the surface of the mast cell results in mast cell degranulation and the release of several proinflammatory mediators. Patients with type-I allergy have increased levels of IgE in the blood compared to healthy individuals. METHODS: In a 6-week culture system of stem cells to human mast cells we investigated the effect of the concentration of IgE. The mast cells were cultured with different concentrations of IgE for the last 10 days of the maturation period. It was observed how the IgE concentration affects the histamine release, FcepsilonRI density on the mast cell surface and the concentration of other mediators. RESULTS: A clear correlation between IgE concentration in culture medium and the release of histamine upon activation was observed. It showed a bell-shaped dose response curve, with maximal response around an IgE-concentration of 250 ng/mL. Furthermore, the sensitivity of the mast cells and surface density of FcepsilonRI on mast cell surface was also influenced by the IgE concentration in the culture medium. CONCLUSIONS: IgE in the culture medium during the last 10 days of mast cell maturation influences the release of the preformed mediator histamine after mast cell activation and the density of FcepsilonRI on the mast cell surface. The release of the de novo synthetized mediator prostaglandin D2 and the expression of chymase and tryptase are not influenced by IgE in culture medium.

The Influence of IgE on Cultured Human Mast Cells

PURPOSE: The mast cell plays a pivotal role in the human immune response. Crosslinking of 2 IgE molecules bound to the high affinity IgE receptor (FcepsilonRI) on the surface of the mast cell results in mast cell degranulation and the release of several proinflammatory mediators. Patients with type-I allergy have increased levels of IgE in the blood compared to healthy individuals. METHODS: In a 6-week culture system of stem cells to human mast cells we investigated the effect of the concentration of IgE. The mast cells were cultured with different concentrations of IgE for the last 10 days of the maturation period. It was observed how the IgE concentration affects the histamine release, FcepsilonRI density on the mast cell surface and the concentration of other mediators. RESULTS: A clear correlation between IgE concentration in culture medium and the release of histamine upon activation was observed. It showed a bell-shaped dose response curve, with maximal response around an IgE-concentration of 250 ng/mL. Furthermore, the sensitivity of the mast cells and surface density of FcepsilonRI on mast cell surface was also influenced by the IgE concentration in the culture medium. CONCLUSIONS: IgE in the culture medium during the last 10 days of mast cell maturation influences the release of the preformed mediator histamine after mast cell activation and the density of FcepsilonRI on the mast cell surface. The release of the de novo synthetized mediator prostaglandin D2 and the expression of chymase and tryptase are not influenced by IgE in culture medium.

Identification of Inducible Murine Mast Cell Genes by Suppression PCR - Based Subtractive Hybridization / 대한면역학회지

The mast cell is an essential effector cell in allergic inflammation through its capacity to respond to IgE dependent activation. Mast cells also participate in the modulation of physiologic processes, but the role of mast cell in these processes is still unclear. Recently, the number of structurally defined chernoattractants for leukocytes has greatly increased, owing to largely to the identification of the chemokine superfamily. In this study we examined the pattern of expression of chemokines and their receptors in HMC-1 after treatment with PMA/A23187 and/or LPS using RT-PCR and ELISA. Messenger RNA of IL-8, the representative CXC chemokine, was induced after PMA/ A23187 treatment. All of the CC chemokines tested, except eotaxin, were induced after PMA/A23187 treatment. CCR1, CXCR2, CXCR3 and CXCR4 were expressed in all test groups regardless of activation. CCR3 was expressed only at 3 hours of activation. CCR2, CCR5 and CXCR1 were not expressed in mast cell line. Production of most of chemokine proteins was not detected in resting state and increased significantly after 3 hours of activation with PMA/A23187. The effect of LPS treatment was negligible. MCP-1 protein was always produced without activation and accurnulated in a time-dependent rnanner. These data suggest that the expression of mRNA and protein of chemokines and chemokine receptors are regulated transcriptionally and translationally. Human mast cell may respond to various stimuli by producing chemokines and their receptors to regulate their function and may act autonomously or through other inflammatory cell that they recruited.

Induction and Expression of Chemokines and Their Receptors in Human Mast Cell Line ( HMC - 1 ) / 대한면역학회지

The mast cell is an essential effector cell in allergic inflammation through its capacity to respond to IgE dependent activation. Mast cells also participate in the modulation of physiologic processes, but the role of mast cell in these processes is still unclear. Recently, the number of structurally defined chernoattractants for leukocytes has greatly increased, owing to largely to the identification of the chemokine superfamily. In this study we examined the pattern of expression of chemokines and their receptors in HMC-1 after treatment with PMA/A23187 and/or LPS using RT-PCR and ELISA. Messenger RNA of IL-8, the representative CXC chemokine, was induced after PMA/ A23187 treatment. All of the CC chemokines tested, except eotaxin, were induced after PMA/A23187 treatment. CCR1, CXCR2, CXCR3 and CXCR4 were expressed in all test groups regardless of activation. CCR3 was expressed only at 3 hours of activation. CCR2, CCR5 and CXCR1 were not expressed in mast cell line. Production of most of chemokine proteins was not detected in resting state and increased significantly after 3 hours of activation with PMA/A23187. The effect of LPS treatment was negligible. MCP-1 protein was always produced without activation and accurnulated in a time-dependent rnanner. These data suggest that the expression of mRNA and protein of chemokines and chemokine receptors are regulated transcriptionally and translationally. Human mast cell may respond to various stimuli by producing chemokines and their receptors to regulate their function and may act autonomously or through other inflammatory cell that they recruited.