Figure 4

Decreased ability of DC-STAMP knock-down mBMDCs to stimulate T cell proliferation in allogeneic MLR. Splenocytes depleted for B220⁺ cells from Balb/c mice were stained with CFSE and co-cultured with immature (-LPS) or LPS-matured (+LPS) DC-STAMP knock-down (shST1) or control (shScr) mBMDCs from C57BL/6 mice. (A) FACS data of CFSE dilution in CD3⁺ T cells. (B) Quantified T-cell proliferation as a mean fluorescent intensity (MFI) of CFSE dilution in CD3⁺ T cells. Mean ± SEM of four independent experiments are shown. (C) FACS data of CD62L surface expression in CD3⁺CD4⁺ and CD3⁺CD8⁺ T cells (Balb/c) from co-cultures with DC-STAMP knock-down (shST1) or with control (shScr) mBMDCs (C57BL/6) (black line) compared to isotype control (grey shaded area). Data shown are representative of three independent experiments. (D) Quantification of CD62L MFI on CD3⁺CD4⁺ and CD3⁺CD8⁺ T cells (Balb/c) from co-cultures with DC-STAMP knock-down mBMDCs (shST1) and shScr mBMDCs (C57BL/6). Data are expressed as mean ± SEM of three independent experiments. (E) Supernatants from allogeneic MLR co-cultures with mature mBMDCs were analyzed for indicated cytokines and chemokines by Milliplex bead assay. Data shown are mean ± SEM from triplicates and are representative of at least three experiments. Two-tailed Student t test was performed in B, D and E (ND-not detectable, *p < 0.05; **p < 0.01; ***p < 0.001).