Graded Zirconia-hydroxyapatite composite bioceramic and simplex Zirconia-hydroxyapatite composite bioceramic were produced, and the extractes of these two of materials were made to evaluate their immunocompatibility. Scanning electron microscope (SEM) was used to detect the character of the surface of the graded composite bioceramic. Holding and without holding phytohemagglutinin (PHA), proliferation and activation of peripheralblood monocytes(PBMCs) cultured in the two extracts were studied. Cultured in PHA after 72 hours, the proliforation rate of the graded composite group was significantly higher than the simplex composite group (P < 0.01). There was no difference of apoptosis of PBMCs of the two groups (P > 0.05). Cultured in PHA after 24 hours, the ratio of CD3/CD69 positive PBMCs of the simple composite group was significantly higher than that of the graded composite material group (P < 0.01). The numbers of PBMCs activated by the graded composite material group were less than that of the simply composite material group and the technique of graded composite will be helpful to improve its immunocompatibility.
Adult ; Cells, Cultured ; Dental Porcelain ; chemistry ; pharmacology ; Durapatite ; chemistry ; pharmacology ; Humans ; Materials Testing ; Monocytes ; drug effects ; immunology ; Zirconium ; chemistry ; pharmacology

Mature dendritic cells are potent antigen-presenting cells that initiate primary immune responses, while immature dendritic cells have quite different properties from mature dendritic cells and are tolerance inducer actually. Here we describe the method of using monocyte condition medium to generate dendritic cells of different maturation phases from nonproliferating progenitors in human peripheral blood. The procedure involves two steps. The first step(or priming phase) is to work on a 6-7-day culture of plastic-adherent blood monocyte in medium supplement with GM-CSF and IL-4. The second step (or differentiation phase) requires the exposure to monocyte conditioned medium. Only the dendritic cells generated by the first step are actually immature, with strong immature dendritic cell features such as active endocytosis, the same expression of monocyte marker CD14, and much of the MHC class II still lies within intracellular compartments (MIIC). The second stage dendritic cells have all the features of mature dendritic cell, including a stellate shape, nonadherence to plastic, the expression of dendritic cells restricted marker CD83, and very strong T cell stimulatory function. All of these dendritic cell properties are retained for at least 3 days when the cytokines are removed, suggesting that these populations are stable and terminally differentiated. Since progression from immature to mature dendritic cell is entirely dependent on exogenously added growth factor such as monocyte condition medium, the peripheral blood monocyte may help to harness synchronized population of mature and immature dendritic cells for studies or therapies.
Cell Culture Techniques ; Cell Differentiation ; Culture Media, Conditioned ; chemistry ; Dendritic Cells ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; chemistry ; Humans ; Interleukin-4 ; chemistry ; Monocytes ; cytology

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

Astragalus membranaceus ; chemistry ; Cell Line ; Cytokines ; metabolism ; Humans ; Inflammation ; Macrophages ; drug effects ; metabolism ; Monocytes ; drug effects ; metabolism ; Polysaccharides ; pharmacology

Adult ; Aged ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Monocytes ; chemistry ; Polymorphism, Genetic ; Thrombomodulin ; blood ; genetics

We analyzed the comparative amounts of granulocyte-colony stimulating factor (G-CSFr) and granulocyte macrophage CSF (GM-CSFr) receptors expressed on neutrophils and monocytes in measles patients to investigate the role of these CSFrs in the development of leukopenia including neutropenia and monocytopenia in measles. EDTA-anticoagulated peripheral blood of 19 measles patients, 10 children with other infections showing leukopenia and 16 children with normal complete blood cell counts (CBC)s were analyzed using flow cytometry and QuantiBRITE. The leukocyte (5260 +/- 2030/uL vs. 9900 + 2680/uL, p=0.000), neutrophil (2580 +/- 960/uL vs. 4250 +/- 2750/uL, p=0.024) and the lymphocyte counts of measles patients (1810 +/- 1430/uL vs. 4530 +/- 3450/uL, p= 0.006) were lower than in the normal controls. The neutrophils of measles patients expressed similar amounts of G- CSFr (1858 +/- 355) as normal children (1764 +/- 477, p= 0.564) and leukopenic patients (1773 +/- 673, p=0.713), but lower levels of GM-CSFr (535 +/- 118) than normal children (957 +/- 344, p=0.000) and leukopenic patients (832 +/- 294, p=0.002). The monocytes of measles patients expressed similar amounts of G-CSFr (916 +/- 336) and GM-CSFr (3718 +/- 906) as normal children (1013 +/- 391 and 4125 (2645, p > 0.05) but less than leukopenic patients (1454 +/- 398 and 5388 +/- 806, p > 0.05). The neutrophil and monocyte counts of measles patients did not correlate with the amount of G-CSFr or GM-CSFr expressed on neutrophils or monocytes (p > 0.05), but in the normal children, the monocyte count correlated with the levels of GM-CSFr on monocytes (r=0.951, p=0.049). In conclusion, neutropenia is one of the more important characteristics of measles patients, which could be due to the decreased GM-CSFr expression on neutrophils. However, the monocytopenia found in measles patients is not due to the decreased expression of CSFr on the monocytes.
Human ; Leukocyte Count ; Measles/*blood ; Monocytes/*chemistry ; Neutropenia/etiology ; Neutrophils/*chemistry ; Receptors, Granulocyte Colony-Stimulating Factor/*blood ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/*blood

OBJECTIVETo study the effect of different concentrations of bilirubin on expression of toll-like receptor 4 (TLR4) in cord blood monocytes (CBMC).

METHODSUnder the sterile condition, umbilical vein blood samples were obtained from normal full-term newborns, and the monocytes were in vitro separated by the method of gelatin/plasma coated flasks. The monocytes were preincubated with various concentrations (0-307.8 μmol/L) of bilirubin dissolved in bovine albumin solution for 1 hr. Bilirubin-treated CBMC were further cultured with LPS (1 μg/mL) to induce cellular activation for 24 hrs, and then the CBMC were collected. The expression of TLR4 in monocytes was measured by indirect immunofluorescence method.

RESULTSBilirubin at the concentrations of 102.6, 153.9, 220.6 and 307.8 μmol/L inhibited the expression of TLR4 of CBMC. The inhibition effect increased with the increasing concentration of bilirubin.

CONCLUSIONSBilirubin can inhibit the TLR4 expression of CBNC in a dose-dependent manner.

Bilirubin ; pharmacology ; Dose-Response Relationship, Drug ; Fetal Blood ; chemistry ; drug effects ; Humans ; Infant, Newborn ; Monocytes ; chemistry ; drug effects ; Toll-Like Receptor 4 ; blood

This study was conducted to get high quality and sufficient numbers of mature dendritic cells from healthy donor peripheral blood. The experiment began on culturing of plastic-adherent monocytes isolated from healthy donor peripheral blood with granulocyte-monocyte clony-stimulating factor (GM-CSF 150 ng/ml) and interleukin 4 (IL-4 800 U/ml) without fresh medium feeding and cytokines for 7 days. After 7 days, CD14+ monocytes not only differentiated into high purity DC but also expressed HLA-I and HLA-II molecules, costimulating molecules, adherent molecules and its progenitor marker CD14 molecule highly. These cells displayed all phenotypic and morphologic characteristics of mature dendritic cells and were most potent stimulatory cells in allogeneic mixed leukocyte reactions. The endocytosis ability of these DCs peaked at the third day in culture and decreased remarkably afterwards. These results provide evidence for the first time that CD14+ monocytes differentiated in vitro from peripheral blood monocytes exhibit dendritic cells characteristics and still express its progenitor marker CD14 molecules highly. The results of this experiments may facilitate further studies of CD14+ DC and its clinical applications.
Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Dendritic Cells ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; chemistry ; Humans ; Interleukin-4 ; chemistry ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; cytology

In our previous work we reported that HIV Tat and 6 cysteine rich peptides of Tat induce tumor necrosis factor-related apoptosis-induced ligand (TRAIL) in human monocytes (Yang et al., 2003). Here our results showed that HIV Tat and Tat cysteine rich peptide increase CCR5 expression in human monocytes, and this activity is inhibited by rabbit anti-Tat. Boiled Tat does not increase CCR5 expression in monocytes. These results provide insight into a new mechanism by which HIV Tat plays a key role in the pathogenesis of HIV-1 infection.
Amino Acid Sequence ; Cells, Cultured ; Cysteine ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Fluid ; chemistry ; Gene Expression Regulation ; drug effects ; physiology ; Gene Products, tat ; chemistry ; pharmacology ; Humans ; Molecular Sequence Data ; Monocytes ; drug effects ; metabolism ; Peptides ; chemistry

BACKGROUNDMacrophage migration inhibitory factor (MIF) plays a pivotal role in inflammatory and immune-mediated diseases. However, its molecular function and role in gastrointestinal diseases has rarely been studied and thus warrants an in-depth investigation. This study was designed and conducted to determine MIF expression in Helicobacter pylori (H. pylori)-induced gastritis and the effect of H. pylori on MIF expression in monocytes in vitro.

METHODSGastric specimens of 62 patients with chronic gastritis were obtained through endoscopic biopsies. Both gastric antrum and body were examined for histopathologic changes. Positive H. pylori was determined through rapid urease test and histopathological examination. A patient was classified as H. pylori positive if both tests showed positive results. The updated Sydney System was employed to assess the severity and activity of gastric inflammation. Double immunoassaying for MIF/T-cells (CD45RO) and MIF/macrophage (KP1), as well as in situ hybridization for the expression of MIF mRNA were used for the current analysis. THP-1, a monocyte cell line, was co-incubated with H. pylori strains (ATCC26695) and subsequently examined for the expression of MIF protein and mRNA by enzyme linked immunosorbent assay and retrospective transcription-polymerase chain reaction, respectively.

RESULTSAmong 62 patients with chronic gastritis, significant increase in total T-cells, MIF+ T-cells, total macrophages, MIF+ macrophages and MIF mRNA+ cells was observed in 42 H. pylori positive patients compared to H. pylori negative patients. Moreover, the increase of the MIF mRNA+ cells was highly correlated with the severity of the disease (number of MIF mRNA+ cells/mm(2), mild: 2834 +/- 382, moderate: 3569 +/- 123, severe: 3881 +/- 118, P < 0.01). In vitro results showed that the expression of MIF protein and mRNA in monocytes was significantly increased after incubation with H. pylori strains.

CONCLUSIONSOverexpression of MIF is common in H. pylori-induced gastric inflammation, which suggests MIF may play an important role in the initiation and development of this disease.

Female ; Gastric Mucosa ; chemistry ; metabolism ; Gastritis ; etiology ; metabolism ; microbiology ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Macrophage Migration-Inhibitory Factors ; analysis ; genetics ; Macrophages ; chemistry ; metabolism ; Male ; Middle Aged ; Monocytes ; metabolism ; RNA, Messenger ; analysis ; T-Lymphocytes ; chemistry ; metabolism