However, no differences were observed (Fig. 1B). The phenotype of the mature moDC was analysed by flow cytometry (Fig. 2). The cells were gated based on size and granularity. Both moDC from immunocompetent Selleckchem Cisplatin controls and immunosuppressed patients with and without previous SCC were CD14 negative and CD1a positive (Fig. 2A and data not shown). Maturation markers MHC class II, CD40, CD80 and CD83 as
well as chemokine receptor CCR7 responsible for homing to lymph nodes were expressed at similar levels. Costimulatory molecule CD86 was expressed at lower levels on moDC from RTR with and without previous SCC compared with controls, but reached only statistical significance in RTR without SCC (P < 0.05). Migration marker CD38 had an increased expression on moDC from RTR (both with and without previous SCC) compared with controls, but statistical significance was only calculated in moDC from patients with previous SCC (P < 0.05 versus controls). When regrouping the RTR according to their immunosuppressive medication, it turned out that both observed differences in surface marker expression were caused by patients treated with a combination of prednisolone
and EPZ-6438 datasheet cyclosporin A or with a combination of prednisolone and azathioprine/mycophenolate mofetil (MMF; Fig. 2B), while patients on triple treatment (prednisolone, cyclosporin A and azathioprine/MMF) showed a similar surface expression of CD86 and CD38 as the immunocompetent controls (Fig. 2B). The supernatants from the moDC populations were tested for cytokine and chemokine production using a 25-plex Luminex
assay. The cytokines IL-2, IL-2R, IL-4, IL-5, IL-6, IL-7, IL-8, IL-12, IL-13, IL-15, IL-17, IFN-α, TNF-α, GM-CSF and chemokines MIP-1β (CCL4), MCP-1 (CCL2), IP-10 (CXCL10), eotaxin (CCL11) and MIG (CXCL9) were produced by moDC from immunosuppressed patients and immunocompetent controls in similar quantities Celecoxib (data not shown). IL-10 and IFN-γ were not detected in the supernatants of neither DC population. The moDC from immunosuppressed patients with previous SCC produced significantly more RANTES (CCL5), MIP-1α (CCL3) and IL-1RA (P < 0.02 versus controls; Fig. 3A). Moreover, the moDC from immunosuppressed patients without SCC produced less IL-1β (both versus controls and immunosuppressed patients with previous SCC). Interestingly, when regrouping the RTR according to their immunosuppressive medication instead of previous history of SCC (Fig. 3B), no difference in IL-1β and RANTES production was observed any longer (data not shown). RTR treated with both prednisolone and cyclosporin A or prednisolone and azathioprine/MMF produced significantly more MIP-1α compared with immunocompetent controls. All treatment groups produced significantly more IL-1RA compared with immunocompetent controls.