Impact of autologous plasma on expression of CD1d, phenotype and function of moDC.

<p><b>a)</b> Representative CD1d expression histograms of moDC cultured in either serum-free AIM-V or RPMI supplemented with either 2% autologous plasma (AP), 10% fetal calf serum (FCS) or 2% delipidated AP. At least three independent experiments were performed. <b>b)</b> Activation of NKT hybridoma cells by alpha-GalCer-loaded moDC generated in either AIM-V or RPMI supplemented with 0.2–1% AP. NKT cell activation was assessed using an IL-2 bioassay. Percentages are relative to the highest activation achieved using 1% AP in the culture medium. Mean and error (SD) are shown. ** p<0.01 as tested by one way ANOVA (Kruskal-Wallis test with Dunn’s post test) <b>c)</b> Proliferation of allogeneic PBMC in a mixed lymphocyte reaction. 10⁵ PBMC were incubated with increasing numbers of moDC. PBMC proliferation was measured by ³H-thymidine uptake. Percentages are relative to the highest cell proliferation observed with 12.5 x 10³ moDC^AIM-V. Mean and error (SD) are shown. <b>d)</b> Representative flow cytometry histograms depicting the expression levels of CD80, CD83, CD86 and HLA-DR on moDC matured overnight with IL-1-beta, IL-6, TNF-alpha and PGE₂. At least three independent experiments were performed. <b>e)</b> Cytokine secretion of LPS/IFN-gamma activated moDC as measured by multiplex assay. Each line represents one individual. V1-V5 denominates the five different individuals tested. Statistical significance was determined using a non-parametric Mann-Whitney test.