Expression of cytokines including IL-6 and tumour necrosis factor-α (TNF-α)21 was increased. Interestingly, transcripts for IL-10, IL-13, interferon-γ (IFN-γ) and IL-12p35 were increased but no production at the protein level was detected.10,21 Furthermore, LPS stimulation did not induce a change in IL-4 gene expression.20 However, T cells that had been exposed to antigen-pulsed MoDCs produced protein

for both IL-4 and IFN-γ.6 In contrast to MoDCs, however, very little information is available on maturation and activation of isolated BDCs following stimulation with LPS. Following their activation and maturation, DCs are known to drive AZD2014 molecular weight T-cell proliferation and to modulate the immune response towards a Th1, Th2, Th17 or T regulatory type of response.1,2 As a result of the limitations of studying T-cell

proliferation in outbred species, most studies in pigs have used mixed lymphocyte reactions6,10,12 and few have used autologous cells.16,21,22 In the present study, both MoDCs and BDCs were isolated from vaccinated pigs and co-cultured with autologous T cells to assess the induction of antigen-specific T-cell activation. We found that both MoDCs and BDCs were equally able to induce T-cell proliferation. However, Ku 0059436 when stimulated with LPS, BDCs that were directly isolated from blood showed a greater increase in cytokine and chemokine expression, when compared with MoDCs. This study therefore provides further evidence that directly isolated BDCs represent an important cell population for studying DC biology in pigs. Further studies, however, are required to identify Acetophenone the specific role of pDCs within the BDC population. Eight-week-old Dutch Landrace pigs purchased from Saskatoon Prairie Swine Centre, University of Saskatchewan were used in this study. The goal of this study was to directly compare MoDCs with isolated BDCs both phenotypically and functionally. Phenotypically, DC morphology was examined by Giemsa staining

and the expression of cell surface markers was examined by flow cytometry. Functionally, endocytic ability was examined by flow cytometry, changes in transcript expression and the production of cytokines in response to stimulation with LPS were investigated using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunsorbent assay (ELISA), respectively, and lastly for their ability to stimulate autologous T-cell proliferation, thymidine uptake assays were performed. Studies were performed as per the ethical guidelines of the University of Saskatchewan and the Canadian Council for Animal Care. Blood was collected by heart puncture using ethylenediaminetetraacetic acid (EDTA) -coated syringes and blood mononuclear cells were isolated using a 60% Ficoll-Paque™ Plus gradient (GE Healthcare, Uppsala, Sweden). Monocytes were isolated using magnetic beads [magnetic antibody cell sorting (MACS); Miltenyi Biotec, Auburn, CA] and human anti-CD14 (TÜK4) microbeads (Miltenyi Biotec).