Complement-induced regulatory T cells suppress T-cell responses but allow for dendritic-cell maturation -- Barchet et al. 107 (4): 1497 Data Supplement - Supplemental materials for: Barchet et al -- Blood

Complement-induced regulatory T cells suppress T-cell responses but allow for dendritic-cell maturation
Blood Barchet et al. 107: 1497

Supplemental materials for: Barchet et al

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Figure S1. Supernatants derived from CD46-induced Tr1-like cells suppress proliferation of bystander T cells through IL-10 (PDF, 12.8 KB) - Purified human CD4⁺ T cells (sorted CD4⁺/CD45RA⁺ cell population) were activated for three days with the indicated immobilized mAbs. As control, T cells were incubated in media with the addition of 10 ng/ml recombinant human IL-10 (rec. IL-10). Supernatants were transferred to freshly purified CD4⁺ T cells and the mixture was incubated with the indicated stimulating antibodies in the presence or absence of function neutralizing mAb to IL-10. Proliferation was measured at day 7 using the CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay from Promega (Madison, WI). This colorimetric assay determines the number of living cells in a sample using an MTS tetrazolium compound (Owen’s reagent). Initial experiments obtained equivalent results using this assay compared to [³H] thymidine incorporation. Aliquots of the same suppressive supernatants were used for the dendritic cell incubation experiments performed in this study. The results are the mean cell proliferation ± SD of three independently performed experiments (with activation conditions in triplicate). *, p < 0.01 by the paired student’s t-test.