This study aims at exploring the effects of tensile strain and loading time on the secretion of IL-1 beta of human periodontal ligament fibroblast. Five tensile strain values including 0%, 8%, 12%, 16%, 20% and three loading time including 24 h, 48 h, 72 h are set in this study. The prepared cell samples are mounted on the self-devised loading apparatus in vitro. The content of IL-1 beta in each sample was determined using double-antibody ELISA. The secretion amount of IL-1 beta per day is directly proportional to the loading time and tensile strain value in tensile strain group of 8%, 12%, 16%. The secretion amount of IL-1 beta reaches its maximum at tensile strain value of 16% in loading time groups. Loaded strain for 24 h and 48 h, the secretion amount of IL-1 beta at tensile strain value of 20% is obviously more than that at value of 0%, but the amount already begin to decrease apparently. Loaded strain for 72 h, the secretion amount of IL-1 beta decreases to a great extent that it is less than the amount at strain value of 0%. The tensile strain stimulates the secretion of IL-1 beta by human periodontal ligament fibroblast when the strain is under normal physiological extent, but the stimulation effect fades out as time goes on.
Cells, Cultured ; Fibroblasts ; secretion ; Humans ; Interleukin-1 ; secretion ; Periodontal Ligament ; cytology ; secretion ; Tensile Strength ; Time Factors

OBJECTIVETo detect the expression of cytokines by acute promyelocytic leukemia (APL) cells before and after exposure to arsenic trioxide.

METHODSDiagnoses were performed according to the FAB cytological classification criteria and cytogenetic criteria. Bone marrow or blood samples from APL patients were collected in heparinized tubes, then primary APL cells were separated by traditional Ficoll-Hypaque density centrifugation and purified after adherence to plastic surfaces. IL-1(beta), IL-6, IL-8, TNF alpha and G-CSF levels in the leukemia cell culture supernatants were detected by ELISA. At the same time, nitro blue tetrazolium (NBT) reduction test was used to detect the differentiation of APL cells.

RESULTSAfter 96 hours exposure to arsenic trioxide, 10 - 6 mol/L in vitro or 10 mg/d in vivo, APL cells showed a significant increase of IL-1(beta) (P < 0.05) and G-CSF (P < 0.05) production, and a significant decrease of IL-6 (P < 0.05) and IL-8 (P < 0.05). However, there was no obvious variation of TNF alpha when compared with APL cells without exposure to arsenic trioxide. On the other hand, the proliferation ratio of APL cells in vitro was statistically correlated to the IL-1(beta) secretion ratio or G-CSF secretion ratio. The cell number ratio in patients with detectable IL-1(beta) or G-CSF was higher than that without detectable IL-1(beta) or G-CSF.

CONCLUSIONIL-1(beta) and G-CSF secretion may play an important role in the proliferation of APL cells after exposure to arsenic trioxide.

Arsenicals ; pharmacology ; Cells, Cultured ; Cytokines ; secretion ; Granulocyte Colony-Stimulating Factor ; secretion ; Humans ; Interleukin-1 ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Leukemia, Promyelocytic, Acute ; metabolism ; Oxides ; pharmacology ; Tumor Necrosis Factor-alpha ; secretion

Atherosclerosis is an inflammatory disease. Platelet-endothelium interaction plays an important role in the pathophysiology of atherogenesis. We investigated the role of activated platelets for secretion of interleukin (IL)-1beta, monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-1alpha and expression of intercellular adhesion molecule (ICAM)-1 on endothelial cells. Human umbilical vein endothelial cells (HUVEC) were incubated with non-stimulated or ADP-activated platelets for 6 hr. Secretion of interleukin (IL)-1beta, MCP-1 and MIP-1alpha and surface expression of ICAM-1 were measured by ELISA and flow cytometry. In the presence of activated platelets, the secretion of IL-1beta, MCP-1, and MIP-1alpha and surface expression of ICAM-1 were significantly increased compared with non-activated platelets. The present study shows that activated platelets may contribute to expression of various inflammatory mediators on endothelial cells.
Blood Platelets/metabolism* ; Cells, Cultured ; Cells, Cultured ; Endothelium, Vascular/secretion* ; Endothelium, Vascular/cytology ; Human ; Intercellular Adhesion Molecule-1/biosynthesis ; Interleukin-1/secretion* ; Macrophage Inflammatory Protein-1/secretion* ; Monocyte Chemoattractant Protein-1/secretion* ; Platelet Activation/physiology*

Atherosclerosis is an inflammatory disease. Platelet-endothelium interaction plays an important role in the pathophysiology of atherogenesis. We investigated the role of activated platelets for secretion of interleukin (IL)-1beta, monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-1alpha and expression of intercellular adhesion molecule (ICAM)-1 on endothelial cells. Human umbilical vein endothelial cells (HUVEC) were incubated with non-stimulated or ADP-activated platelets for 6 hr. Secretion of interleukin (IL)-1beta, MCP-1 and MIP-1alpha and surface expression of ICAM-1 were measured by ELISA and flow cytometry. In the presence of activated platelets, the secretion of IL-1beta, MCP-1, and MIP-1alpha and surface expression of ICAM-1 were significantly increased compared with non-activated platelets. The present study shows that activated platelets may contribute to expression of various inflammatory mediators on endothelial cells.
Blood Platelets/metabolism* ; Cells, Cultured ; Cells, Cultured ; Endothelium, Vascular/secretion* ; Endothelium, Vascular/cytology ; Human ; Intercellular Adhesion Molecule-1/biosynthesis ; Interleukin-1/secretion* ; Macrophage Inflammatory Protein-1/secretion* ; Monocyte Chemoattractant Protein-1/secretion* ; Platelet Activation/physiology*

OBJECTIVES: To investigate the possible role of mononuclear cells and their products in the pathogenesis of IgA nephropathy, in vitro expression of ICAM-1 on cultured mouse mesangial cell (MC) was examined after stimulation with mononuclear cell culture supernatant from patients with IgA nephropathy. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated and cultured from 18 patients with primary IgA nephropathy, 8 normal controls and 5 patients with non-IgA nephropathy (FSGS 1, MGN 3, MPGN 1). ICAM-1 expression on cultured mouse MC by TNF-alpha, IL-1 beta and culture supernants of PBMC were analyzed using a cell ELISA method. The concentration of IL-1 beta and TNF-alpha in culture supernatants was measured by using a commercially available radioimmunoassay kit. RESULTS: Addition of human recombinant TNF-alpha induced an increased ICAM-1 expression in a dose-dependent manner. The expression of ICAM-1 was further increased after co-stimulation with TNF-alpha and IL-1 beta. Addition of PBMC culture supernatants into mouse MC induced significantly higher expression of ICAM-1 by supernatants from the patients with IgA nephropathy compared with that from normal controls. The concentration of TNF-alpha and IL-1 beta in supernatants from the patients with IgA nephropathy was significantly higher than that from those with non-IgA nephropathy. CONCLUSION: TNF-alpha and IL-1 released from mononuclear cells induced the up-regulation of ICAM-1 expression and this may be related to the immune pathogenesis of IgA nephropathy.
Animal ; Cells, Cultured ; Glomerular Mesangium/immunology ; Glomerular Mesangium/cytology ; Glomerulonephritis, IGA/immunology* ; Glomerulonephritis, IGA/etiology ; Human ; Intercellular Adhesion Molecule-1/metabolism* ; Interleukin-1/secretion ; Interleukin-1/pharmacology ; Leukocytes, Mononuclear/immunology ; Mice ; Tumor Necrosis Factor/secretion ; Tumor Necrosis Factor/pharmacology

OBJECTIVETo determine the clinical significance of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha( TNF-alpha) in expressed prostatic secretions(EPS) for chronic prostatitis.

METHODSProstatic secretions IL-1beta and TNF-alpha were evaluated for 34 patients with chronic prostatitis, 10 with asymptomatic inflammatory prostatitis, 12 with benign prostatic hyperplasia (BPH) and 8 health controls by enzyme-linked immunosorbent assay (ELISA).

RESULTSIL-1beta and TNF-alpha levels in EPS in the patients of chronic prostatitis with WBC > or = 10/HP and asymptomatic inflammatory prostatitis were obviously higher than those of chronic prostatitis with WBC < 10/HP, BPH and health controls, (P < 0.05 and P < 0.02). There was a correlation between IL-1beta and TNF-alpha (P < 0.003) but none between WBC and IL-1beta or TNF-alpha.

CONCLUSIONCytokines are frequently elevated in EPS in men of chronic prostatitis with high WBC and asymptomatic inflammatory prostatitis, which provides a novel means different from traditional methods based on WBC for the identification of men with chronic prostatitis.

Aged ; Chronic Disease ; Humans ; Interleukin-1 ; analysis ; Male ; Middle Aged ; Prostate ; chemistry ; secretion ; Prostatitis ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20 positive primary chronic lymphocytic leukemia (CLL) cells and provide a promising tool for tumor adoptive immunotherapy.

METHODSThe recombinant vectors were transduced into PA 317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection for one week. Then transduced T lymphocytes and primary CLL cells were co-cultured. The status of primary chronic lymphocytic leukemia cells were observed by microscope. The level of IL-2 and IFN-gamma in the culture medium were measured.

RESULTSPrimary T cells expressing anti-CD20scFv/IgGFc/CD80/CD28/zeta could be constructed successfully. These T cells were able to lyse CD20+ targets and secrete high levels of IL-2 (1301.00 pg/ml) and IFN-gamma (602.18 pg/ml) in vitro.

CONCLUSION(1) Recombinant gene modified T cells can be constructed successfully. (2) Recombinant gene modified T cells can specially kill CD20 positive primary CLL cells in vitro.

Antigens, CD20 ; genetics ; B7-1 Antigen ; genetics ; CD28 Antigens ; genetics ; Genetic Vectors ; Humans ; Immunotherapy, Adoptive ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Leukemia, Lymphocytic, Chronic, B-Cell ; pathology ; Retroviridae ; genetics ; T-Lymphocytes ; immunology ; secretion ; Transfection ; Tumor Cells, Cultured

Animals ; Cells, Cultured ; Interleukin-1 ; pharmacology ; Matrix Metalloproteinase 1 ; secretion ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Platelet-Derived Growth Factor ; pharmacology ; Rabbits

This study was aimed to prepare and identify human monoclonal antibody against IL-1 receptor accessory protein (IL1RAP), which is a new identified surface marker for leukemia stem cell (LSC), BALB/c mice were immunized with recombinant hu-IL1RAP and the spleen cells from immunized mice were fused with SP2/0 myeloma cells by conventional hybridoma technique. Positive hybridoma cells were selected and cultured. ELISA and Western blot were used to detect the type, titer and sensitivity of antibody. Peripheral blood mononuclear cells were isolated and used to test the antibody specificity. The results showed that 8 hybridoma cell lines able to stably secrete IL1RAP monoclonal antibodies were obtained and named 3H6E10, 4B6A6, 8G11B5, 9E9F2, 10D8A7, 1C7H7, 1D7G11 and 2D3D3 respectively. These monoclonal antibodies belonging to IgG1/κ type could specifically bind to IL1RAP from peripheral blood mononuclear cells. It is concluded that the hybridoma cell lines with stable secretion of IL1RAP monoclonal antibodies is successfully constructed, thus providing novel ways to effectively clear LSC in the future.
Animals ; Antibodies, Monoclonal ; analysis ; Antibody Specificity ; immunology ; Cell Line ; Humans ; Hybridomas ; immunology ; secretion ; Interleukin-1 Receptor Accessory Protein ; immunology ; Mice ; Mice, Inbred BALB C ; Neoplastic Stem Cells ; immunology

This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25 - 25 mmol.L-1 sodium nitrite for 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-beta1, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-beta1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol.L-1 of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-beta1 and IL-8.
Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Liver Neoplasms ; metabolism ; pathology ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-akt ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Transforming Growth Factor beta1 ; secretion ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism