Cutaneous dendritic cells (DCs), Langerhans cells (LCs) and dermal dendritic cells (DDCs), are present in an immature state. The maturation of DCs is crucial for initiating an immune response. Since HLA-DM has an important role for antigen presentation, an increase in HLA-DM expression according to the maturation of blood monocyte-derived dendritic cells (MoDCs), which have similar characteristics with DDCs, is expected. Therefore, the aim of this study was to determine whether or not HLA-DM expression in MoDCs is related to maturation at each culture day (from day 0 to day 13) by flow cytometry. This was compared with the functional changes related to the maturation of MoDCs. MoDCs were generated by culturing human peripheral blood monocytes in the presence of GM-CSF and IL-4 for 7 days, which were followed by subsequent treatment with a cytokine cocktail (GM-CSF, IL-4, IL-1beta, TNF-alpha, IL-6 and PGE2) for the maturation of MoDCs. The intracellular HLA-DM was expressed in the immature MoDC. A sudden 3 to 8 fold increase in the intracellular HLA-DM expression was observed after treatment with a cytokine cocktail. HLA-DM was weakly expressed on the surface of the immature MoDC, but it seemed to be decreased with maturation. This study indicated that the intracellular HLA-DM expression increased, but not on the MoDC surface during maturation. This was despite the fact that HLA-DM expression was noted not only on the surface but also in the intracellular in the MoDC.
Dendritic Cells/*immunology/physiology ; Endocytosis ; Flow Cytometry ; HLA-D Antigens/*analysis ; Human ; Monocytes/*physiology

The immunocytes microglia in the central nervous system (CNS) were reported to play a crucial role in neurodegeneration. As a member of P2 receptors family, purinoceptor P2Y6 has attracted much attention recently. Previous studies showed that purinoceptor P2Y6 mainly contributed to microglia activation and their later phagocytosis in CNS, while in immune system, it participated in the secretion of interleukin (IL)-8 from monocytes and macrocytes. So there raises a question: whether purinoceptor P2Y6 also takes part in neuroinflammation? Thus, this review mainly concerns about the properties and roles of purinoceptor P2Y6, including (1) structure of purinoceptor P2Y6; (2) distribution and properties of purinoceptor P2Y6; (3) relationships between purinoceptor P2Y6 and microglia; (4) relationships between purinoceptor P2Y6 and immunoinflammation. Itos proposed that purinoceptor P2Y6 may play a role in neuroinflammation in CNS, although further research is still required.
Animals ; Humans ; Inflammation ; immunology ; metabolism ; Microglia ; drug effects ; metabolism ; Monocytes ; metabolism ; Phagocytosis ; physiology ; Receptors, Purinergic P2 ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate the relationship between aging and the number and function of bone marrow-derived endothelial progenitor cells (EPC) in rats.

METHODSThe number and function of bone marrow-derived EPC were measured in 4, 10 and 18 months old rats (n = 10 in each group). Mononuclear cells (MNC) were isolated from rat bone marrow by Ficoll density gradient centrifugation and cultured for 7 days, EPC were identified as adherent cells double positive stained for FITC-UEA-I and DiI-acLDL under laser confocal immunofluence microscopy. Boyden chamber was used to determine the migration capacity of EPC and the number of recultured EPC was counted to test the adherent function of EPC.

RESULTThe number (42.0 +/- 5.8 vs 30.8 +/- 4.7 vs 21.5 +/- 4.7 per field) and the migration and adhesion capacities of EPC were significantly reduced with aging.

CONCLUSIONThis study shows that the number and function of EPC are decreased with aging.

Age Factors ; Animals ; Bone Marrow Cells ; cytology ; Cell Count ; Cells, Cultured ; Endothelial Cells ; cytology ; Monocytes ; physiology ; Rats ; Rats, Sprague-Dawley

In order to determine the precise mechanism of the interactions between different types of cells, which are common phenomena in tissues and organs, the importance of coculture techniques are becoming increasingly important. In the area of cardiology, artificial arteries have been developed, based on the understanding of physiological communication of the arterial smooth muscle cells (SMC), endothelial cells (EC), and the extracellular matrix (ECM). In the study of atherosclerosis, the modification of low-density lipoprotein (LDL), which result in the recruitment and accumulation of white blood cells, especially, monocytes/macrophages, and foam cell formation, are hypothesized. Although there are well known animal models, an in vitro model of atherogenesis with a precisely known atherogenesis mechanism has not yet been developed. In this paper, an arterial wall reconstruction model using rabbit primary cultivated aortic SMCs and ECs, was shown. In addition, human peripheral monocytes were used and the transmigration of monocytes was observed by scanning electron and laser confocal microscopy. Monocyte differentiation into macrophages was shown by immunohistochemistry and comprehensive gene expression analysis. With the modified form of LDL, the macrophages were observed to accumulate lipids with a foamy appearance and differentiate into the foam cells in the ECM between the ECs and SMCs in the area of our coculture model.
Animal ; Aorta/physiology* ; Aorta/cytology* ; Arteriosclerosis/etiology* ; Cell Differentiation/physiology ; Cell Movement ; Coculture ; Endothelium, Vascular/physiology ; Endothelium, Vascular/cytology ; Extracellular Matrix/metabolism ; Foam Cells/ultrastructure ; Foam Cells/cytology ; Macrophages/physiology* ; Macrophages/cytology ; Male ; Microscopy, Confocal ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Monocytes/ultrastructure ; Monocytes/physiology ; Muscle, Smooth, Vascular/physiology ; Muscle, Smooth, Vascular/cytology ; Myosin/metabolism ; Protein Isoforms/metabolism ; Rabbits

It is generally accepted that tissue factor plays an important role in coagulation and intravascular thrombus formation. Tissue factor is not only found primarily on the surface of certain cells that are located outside the vasculature, but also found in circulating cells. Monocyte express tissue factor induced by endotoxin. Recently, many researches indicate that P-selectin, CD40 ligand and GPIIb/IIIa receptor of platelet can also affect expression of tissue factor by monocyters. In addition, a lot of studies showed that tissue factor exist in the circulation including contained platelet. Tissue factor in the platelet releases under certain condition, and initiates coagulation. In this review the relation between platelet and tissue factor was elaborated.
Blood Platelets ; drug effects ; metabolism ; physiology ; CD40 Ligand ; physiology ; Humans ; Monocytes ; drug effects ; metabolism ; P-Selectin ; physiology ; Peptides ; pharmacology ; Platelet Glycoprotein GPIIb-IIIa Complex ; physiology ; Thromboplastin ; biosynthesis ; drug effects ; physiology

AIMTo observe the property alteration of K(v) channel in SLE patient's peripheral blood lymphocyte and its significant.

METHODSThe patch-clamp technique was used to record the current of K(V) channel in SLE patient's peripheral lymphocyte.

RESULTSThe current amplitude of K(V) channel in the SLE patient's lymphocytes decreased, it was (258.6 +/- 112.5) pA in healthy people, but in SLE patient it was (139.4 +/- 58.5) pA (P < 0.05). There was no other changes in the property of channel, include activation potential, inactivation property, channel closing kinetics and its pharmacological property.

CONCLUSIONThe decline of SLE patient's cell immunity may be related to the decrease of the amplitude of K(V) channel current.

Case-Control Studies ; Cell Membrane ; physiology ; Flow Cytometry ; Humans ; Lupus Erythematosus, Systemic ; blood ; pathology ; physiopathology ; Lymphocyte Count ; Lymphocytes ; physiology ; Monocytes ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; physiology

Oxidized LDL (OxLDL), a causal factor in atherosclerosis, induces the expression of heat shock proteins (Hsp) in a variety of cells. In this study, we investigated the role of CD36, an OxLDL receptor, and peroxisome proliferator-activated receptor gamma (PPAR gamma) in OxLDL-induced Hsp70 expression. Overexpression of dominant-negative forms of CD36 or knockdown of CD36 by siRNA transfection increased OxLDL-induced Hsp70 protein expression in human monocytic U937 cells, suggesting that CD36 signaling inhibits Hsp70 expression. Similar results were obtained by the inhibition of PPAR gamma activity or knockdown of PPAR gamma expression. In contrast, overexpression of CD36, which is induced by treatment of MCF-7 cells with troglitazone, decreased Hsp70 protein expression induced by OxLDL. Interestingly, activation of PPAR gamma through a synthetic ligand, ciglitazone or troglitazone, decreased the expression levels of Hsp70 protein in OxLDL-treated U937 cells. However, major changes in Hsp70 mRNA levels were not observed. Cycloheximide studies demonstrate that troglitazone attenuates Hsp70 translation but not Hsp70 protein stability. PPAR gamma siRNA transfection reversed the inhibitory effects of troglitazone on Hsp70 translation. These results suggest that CD36 signaling may inhibit stress- induced gene expression by suppressing translation via activation of PPAR gamma in monocytes. These findings reveal a new molecular basis for the anti-inflammatory effects of PPAR gamma.
Antigens, CD36/*physiology ; Cell Line, Tumor ; Chromans/pharmacology ; Cycloheximide/pharmacology ; HSP70 Heat-Shock Proteins/*biosynthesis ; Humans ; Lipoproteins, LDL/pharmacology/*physiology ; Monocytes/drug effects/metabolism ; PPAR gamma/agonists/antagonists & inhibitors/*physiology ; Protein Synthesis Inhibitors/pharmacology ; Signal Transduction ; Thiazolidinediones/pharmacology

Atherosclerosis ; etiology ; prevention & control ; Cytokines ; secretion ; Dendritic Cells ; drug effects ; immunology ; Endocytosis ; Fenofibrate ; pharmacology ; Humans ; Immunophenotyping ; Lipoproteins, LDL ; toxicity ; Monocytes ; cytology ; PPAR alpha ; agonists ; physiology

Adolescent ; Adult ; Aged ; Female ; HLA-DR Antigens ; blood ; Humans ; Lipopolysaccharide Receptors ; blood ; Male ; Middle Aged ; Monocytes ; physiology ; Severe Acute Respiratory Syndrome ; immunology

To investigate the effects of DENV infection on the expression of Vascular endothelial growth factor (VEGF) in monocytes, and to explore which innate immune signaling pathway is responsible for VEGF induction. Real-time PCR was used to determine the expression levels of VEGF in DENV-infected THP-1. We found that different serotype viruses (DENV1, DENV2, DENV3) induced the VEGF expression. Moreover, VEGF expression was significantly increased in human primary monocytes infected with DENV 2. In addition, VEGF induction by DENV2 was significantly impaired by knockdown of TLR3 and interferon-beta promoter stimulator 1 (IPS-1), or by inhibition of ERK, JNK or NF-kappaB. These results demonstrated that DENV induced VEGF expression in monocytes, and the activation of TLR3, IPS-1 signal pathways were required for DENV2-triggered VEGF induction, suggesting that VEGF might be a promising therapeutic target for DHF.
Dengue ; genetics ; immunology ; virology ; Dengue Virus ; classification ; genetics ; physiology ; Humans ; MAP Kinase Signaling System ; Monocytes ; NF-kappa B ; genetics ; immunology ; Up-Regulation ; Vascular Endothelial Growth Factor A ; genetics ; immunology