Comparison of the innate immune responses of porcine monocyte-derived dendritic cells and splenic dendritic cells stimulated with LPS

Abstract

Dendritic cell (DC) subsets form a remarkable cellular network that regulate innate and adaptive immune responses. Although pigs are the most approximate model to humans, little is known about the regulation of monocyte-derived DCs (moDCs) and splenic DCs (SDCs) in the initiation of immune responses under inflammatory conditions. We investigated the activation and maturation of porcine moDC and SDC subpopulations following LPS stimulation. Porcine monocytes that would differentiate into moDCs were isolated. SDCs were isolated directly from the porcine spleen. Following LPS stimulation, phagocytosis activity, TLR4/MyD88-dependent gene expression, co-stimulatory molecule, and pro-inflammatory cytokine (TNF-α, IL-1β) and chemokine (IL-8) expressions were increased in both cell subsets. Furthermore, moDCs showed higher levels of gene and protein expression compared with SDCs. Interestingly, moDCs were found to be more responsive via the TLR4/TRAF-dependent signalling pathway of activation. Only SDCs expressed higher level of IL-12p40 gene and protein, whereas, IFN-γ gene and protein expression were likely to be unchanged after LPS stimulation in both cell subtypes. These data demonstrate that porcine moDCs display a greater ability to initiate innate immune responses, and could be used as a model to investigate immune responses against Ags.

The role of toll-like receptor 4 in tumor microenvironment

“…Among them, the expression of TLR4 is restricted to mDCs. Following LPS stimulation, DCs separated from pigs presented an enhanced TLR4/MyD88-dependent gene expression, CD40 and CD86 on the cell surface, as well as chemokine and pro-inflammatory cytokine expressions [ 25 ] (Figure 1 ). Mature DCs are able to induce T helper 1 (TH1) -type immune responses, and are also viewed as potent initiators of tumor associated antigen (TAA)-specific immunity.…”

Characterization of polarization states of canine monocyte derived macrophages

“…Pan-macrophage markers CD14 and CD11b [30] were highly expressed indicating that these cells indeed maintain macrophage properties during polarization [31]. Co-stimulatory molecules CD40, CD80 and CD86 together with MHC II are essential for antigen presentation and T cell interaction by macrophages and are considered as M1 macrophage markers in many species [31][32][33][34]. MHC class II showed increased expression in both M1 and M2 polarized cells.…”

Epigenetic Modulation of TLR4 Expression by Sulforaphane Increases Anti-Inflammatory Capacity in Porcine Monocyte-Derived Dendritic Cells

“…DCs were derived from cultured porcine monocytes isolated from peripheral blood monocular cells (PBMCs) following the protocol described previously [15,17]. Briefly, PBMCs were isolated from two porcine peripheral blood samples using Ficoll-Histopaque medium (cat.…”

“…At the end of the PCR, a melting curve analysis was performed to detect the specificity of the PCR. Details of the PCR conditions have been described previously [15,17]. Each experiment was performed in triplicate and each sample was quantified in duplicate (technical replication).…”

“…80004; QIAGEN). Equal amounts of cell lysates were loaded and electrophoresed through precast gels, transferred onto nitrocellulose membranes, and confirmed with ponceau S staining [17]. Membranes were incubated with primary antibodies specific for anti-acetylated histones H3 (H3-Ac) (cat.…”