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• W2352416175 abstract "Objective To investigate the effect of buthionine sulfoximine(BSO),a glutathione(GSH) synthesis inhibitor,on differentiation and maturation of human dendritic cells(DCs). Methods Monocytes isolated from peripheral blood monuclear cells(PBMCs) of health donors were cultured with GM-CSF and IL-4 in RPMI 1640 medium to produce DCs. The monocytes were divided into early treatment(added with BSO 1 mmol/L from day 1) late treatment(added with BSO 1 mmol/L from day 5),and control(cultured without BSO) groups. The medium was changedevery other day with the BSO complemented to a same concentration(1 mmol/L). On day 7,the unattached cells were collected to analyze their phenotypes and function;the attached cells in the early treatment and control groups were collected and observed under an inverted microscope. In another experiment,the unattached cells were transferred to a new plate on day 5,cultured with LPS(100 ng/ml),IFN-γ(20 U/ml),or without any stimulation for 48 hours,and then collected for phenotyping with specific monoclonal antibodies and flow cytometry. In the functional assay,mitomycin C-treated DCs were co-cultured with allogeneic unattached PBMCs(1×105) in ratios of 1:25,1:50,or 1:100 for 5 days. The proliferation of the lymphocytes was analyzed by measuring the value of 3H-thymidine incorporation with β-counter. Results On day 7,the attached cells in the early treatment group was significantly more than that in the control group. Flow cytometry showed that the CD86+ and CD1a+ cells in the early treatment group were less than those in the control group,mean fluorescence intensity(MFI) value of CD86 was 311.3 ± 97.3 and 552.0 ± 97.9(P = 0.01),while MFI value of CD1a was 52.2 ± 22.2 and 121.2 ± 4.2(P = 0.03),respectively. The value of 3H-thymidine incorporation in the early treatment group was significantly lower than that in the control group when the ratio of DCs to nonadherent PBMCs was 1:25(1587 ± 1458 vs. 9336 ± 1333,P = 0.015). After being cultured with IFN-γ(20 U/ml),the MFI value of CD86 in the early treatment and control groups was 495.9 ± 143.9 and 800.4 ± 27.7,respectively,while it was 1736.7 ± 375.7 and 1960.8 ± 494.5(P 0.05) in those treated with LPS(100 ng/ml),indicating that both the differentiation of monocytes into DCs and IFN-γ-induced maturation of DCs was inhibited by the early treatment with BSO. No significant difference was detected in the expression of the antigen presenting-related molecules among the cells treated with LPS,IFN-γ,or none of them in the late treatment group,and the control group. Conclusions Early treatment with BSO could inhibit the differentiation of monocytes into DCs and the maturation of DCs. Cellular redox state at early stage might be a pivotal decision factor for the differentiation and maturation of DCs." @default.
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• W2352416175 date "2007-01-01" @default.
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• W2352416175 title "Early treatment with buthionine sulfoximine inhibits differentiation of monocytes into dendritic cells" @default.
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