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- W2034876444 abstract "RATIONALE: We have addressed the in vivo and ex vivo significance of sphingosine kinase (SPHK) inhibition in Immunoglobulin E (IgE)-mediated allergic responses. METHODS: The in vivo studies were performed on male BALB/c mice, aged 10 weeks obtained from the Laboratory Animal Holding Unit, National University of Singapore, Singapore. To induce PCA, the BALB/c mice were lightly anesthetized and injected intradermally on the basolateral side with 20 ng/ml of monoclonal mouse anti-DNP IgE diluted in 20 μl of saline. Saline alone was used as the negative control; the positive control group was given only the anti-DNP-IgE without N, N-Dimethyl D-erythro-sphingosine (DMS). For in vitro studies, total bone marrow samples from femurs of 3-5 mice were cultured in RPMI medium supplemented with 10% FCS, 10 ng/ml IL3, 20 ng/ml SCF, 2 mM L-glutamine, 100 U/ml penicillin and streptomycin in a 5% CO2 atmosphere. After a 4-week maturation period, BMMCs were collected and tested for c-kit receptor and FcεRI expression by flow cytometry using phycoerythrin-labelled mouse anti c-kit (Pharmigen), or anti-DNP IgE (SPE-7; Sigma) and mAb fluorerscin anti-mouse IgE (Molecular probes) antibodies, respectively. RESULTS: Injection of DMS subcutaneously, or intravenously, inhibited the PCA response in vivo. Moreover, inhibition of SPHK in BMMC, by either DMS or specific SPHK 1-siRNA, decreases several effector function, normally triggered during FcεRI aggregation, such as degranulation, the release of proinflammatory cytokines (IL-6, IL-3 and TNF α), as well as the de novo synthesis of the arachidonic acid metabolites such as leukotrienes, and prostaglandins. CONCLUSIONS: Our data suggest that SPHK is a potential key mediator of IgE-mediated anaphylactic responses, and other mast cell related pathologies." @default.
- W2034876444 created "2016-06-24" @default.
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- W2034876444 date "2006-02-01" @default.
- W2034876444 modified "2023-09-27" @default.
- W2034876444 title "Essential Role of Sphingosine Kinase in IgE-mediated Allergic Response in vivo and ex vivo" @default.
- W2034876444 doi "https://doi.org/10.1016/j.jaci.2005.12.538" @default.
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