Stimulatory Interactions between Human Coronary Smooth Muscle Cells and Dendritic Cells

Abstract

Despite inflammatory and immune mechanisms participating to atherogenesis and dendritic cells (DCs) driving immune and non-immune tissue injury response, the interactions between DCs and vascular smooth muscle cells (VSMCs) possibly relevant to vascular pathology including atherogenesis are still unclear. To address this issue, immature DCs (iDCs) generated from CD14+ cells isolated from healthy donors were matured either with cytokines (mDCs), or co-cultured (ccDCs) with human coronary artery VSMCs (CASMCs) using transwell chambers. Co-culture induced DC immunophenotypical and functional maturation similar to cytokines, as demonstrated by flow cytometry and mixed lymphocyte reaction. In turn, factors from mDCs and ccDCs induced CASMC migration. MCP-1 and TNFα, secreted from DCs, and IL-6 and MCP-1, secreted from CASMCs, were primarily involved. mDCs adhesion to CASMCs was enhanced by CASMC pre-treatment with IFNγ and TNFα ICAM-1 and VCAM-1 were involved, since the expression of specific mRNAs for these molecules increased and adhesion was inhibited by neutralizing antibodies to the counter-receptors CD11c and CD18. Adhesion was also inhibited by CASMC pre-treatment with the HMG-CoA-reductase inhibitor atorvastatin and the PPARγ agonist rosiglitazone, which suggests a further mechanism for the anti-inflammatory action of these drugs. Adhesion of DCs to VSMCs was shown also in vivo in rat carotid 7 to 21 days after crush and incision injury. The findings indicate that DCs and VSMCs can interact with reciprocal stimulation, possibly leading to perpetuate inflammation and vascular wall remodelling, and that the interaction is enhanced by a cytokine-rich inflammatory environment and down-regulated by HMGCoA-reductase inhibitors and PPARγ agonists.

PARP-1 Is Critical for Recruitment of Dendritic Cells to the Lung in a Mouse Model of Asthma but Dispensable for Their Differentiation and Function

“…Expression of VCAM-1 on structural cells such as those of the smooth muscle also influences DC localization in inflamed tissues [40] and participates in tissue remodeling [40, 41]. We thus examined whether the effect of PARP-1 gene knockout on DC migration to the lung was associated with a reduction of VCAM-1 expression in lungs of OVA-sensitized and OVA-challenged mice.…”

Vaginal Lactobacilli and Vaginal Dysbiosis-Associated Bacteria Differently Affect Cervical Epithelial and Immune Homeostasis and Anti-Viral Defenses

“…Buffy coats from healthy donors ( n = 9) were supplied by the Transfusional Center of Azienda Ospedaliera Careggi (Firenze, Italy). PBMCs were isolated by Ficoll-Paque density gradient (Cedarlane Labs, Burlington, Ontario, Canada) according to Paccosi et al [ 53 ] and cultured in 6-well plates at the concentration of 10 6 cells/mL in RPMI-1640 medium (Euroclone, Pero, Italy) supplemented with 10% FBS, 1% L-glutamine and 1% penicillin and streptomycin (Euroclone, Pero, Italy). After 1 h at 37 °C, cells were stimulated with heat-inactivated bacteria (50 MOI/cell) and cultured for additional 5 days at 37 °C and 5% di CO 2 .…”

Differentiation of Human Hematopoietic Progenitor Cells Into Dendritic Cells In Vitro Under Stimulation of PPAR-Gamma

“…Human buffy coats were stored at +4 °C for 24 h before use and then subjected to density gradient centrifugation on Ficoll (Lymphoprep, Euroclone, Pero, Italy) followed by immunomagnetic separation of CD14+ cells, following a previously published protocol [12,29]. In detail, the buffy coat was suspended in 2,5% dextran (Amersham Pharmacia Biotech, Sweden) in 0.9% NaCl and stabilized at 37 °C for 30 min.…”

“…In vitro, DCs can differentiate from circulating CD14+ and CD14-monocytes [10][11][12], from CD34+ progenitors and from CD133+ precursors; the last cells have been used only in a few studies [13][14][15]. Attempts to drive the differentiation towards specific subtypes of DCs, in particular towards LCs, have given inconsistent results, the best ones have been obtained starting from CD14-cells of adult peripheral blood [11] and from CD34+ [16] or CD133+ precursors isolated from cord blood [15].…”