Figure 4

RA generated CD11b⁺BM-MCs suppress T cell proliferation and express a regulatory phenotype. CD11b⁺ CD11c^- were purified from BM-MCs by sequential purification of CD11b⁺ cells from the CD11c- fraction and purity determined to be >95%. Cells were routinely stained for cell surface markers and flow cytometry performed using the BD Accuri C6 Flow cytometer. CD11b⁺ CD11c cells were (A) assessed for expression of maturation markers CD80, CD86, MHCII, PD-L1 and PD-L2 (B) co-cultured with responder immune cells for 96 hours with media or anti-CD3 stimulation and then pulsed with H³ thymidine in the final 18 hours of culture, (C) intracellular cytokine staining (IL-17, INF-γ, IL-4) of CD4⁺ cells performed and (D) co-cultured with activated CTLs and a T lymphoma cell line with cell viability assessed by the MTT assay. Data is representative of three separate experiments. * = p < 0.05 (A-C) Data shown is a representation of 3 experiments (D).