TNF-related apoptosis inducing ligand (TRAIL) expressions were studied in primary human brain astrocytes in response to pro-inflammatory cytokines. When astrocytes were treated with IL-1beta TNF-alphaor IFN-gamma TRAIL was induced in cultured fetal astrocytes. In particular, IFN-gammainduced the highest levels of TRAIL in cultured astrocytes. When astrocytes were pre-reated with IFN-gamma they induced apoptosis in TRAIL-sensitive Peer cells. Our results suggest that IFN-gamma modulates the expression of TRAIL in astrocytes, which may enhance cytotoxic sensitivity of infiltrating immune cells or brain cells other than astrocytes during inflammation of brain.
Tumor Necrosis Factor-alpha/genetics/*metabolism ; TNF-Related Apoptosis-Inducing Ligand ; Membrane Glycoproteins/genetics/*metabolism ; Interferon Type II/*pharmacology ; Humans ; Cells, Cultured ; Astrocytes/*cytology/drug effects/metabolism ; Apoptosis Regulatory Proteins/genetics/*metabolism ; Apoptosis/*drug effects/physiology ; Antineoplastic Agents/*pharmacology

TNF-related apoptosis inducing ligand (TRAIL) expressions were studied in primary human brain astrocytes in response to pro-inflammatory cytokines. When astrocytes were treated with IL-1beta TNF-alphaor IFN-gamma TRAIL was induced in cultured fetal astrocytes. In particular, IFN-gammainduced the highest levels of TRAIL in cultured astrocytes. When astrocytes were pre-reated with IFN-gamma they induced apoptosis in TRAIL-sensitive Peer cells. Our results suggest that IFN-gamma modulates the expression of TRAIL in astrocytes, which may enhance cytotoxic sensitivity of infiltrating immune cells or brain cells other than astrocytes during inflammation of brain.
Tumor Necrosis Factor-alpha/genetics/*metabolism ; TNF-Related Apoptosis-Inducing Ligand ; Membrane Glycoproteins/genetics/*metabolism ; Interferon Type II/*pharmacology ; Humans ; Cells, Cultured ; Astrocytes/*cytology/drug effects/metabolism ; Apoptosis Regulatory Proteins/genetics/*metabolism ; Apoptosis/*drug effects/physiology ; Antineoplastic Agents/*pharmacology

BACKGROUND: Mycobacterial antigens released as PIM, LM, LAM, lipoproteins and other cellular factors may contribute to macrophage and dendritic cell activation through pattern recognition receptors such as TLRs. In this study, we assessed cytokine production and ERK activation with stimulation of several major mycobacterial antigens. METHODS: Purified mycobacterial antigens (10, 22, 30, 38kappaDa) and recombinant antigens (6, 16, 19, 38kappaDa, Ag85A antigen) were studied. The production of cytokines (TNF-alpha, IL-12, IL-6) was measured by ELISA. The ERK activation was detected by western blotting. The expression of TLR2 or TLR4 was measured by flow cytometry. RESULTS: Among purified antigens only 30kappaDa antigen induced production of IL-6 or TNF-alpha in THP-1 macrophage cells. When THP-1 macrophage cells were treated with 30kappaDa antigen, phosphorylation of ERK was detected. ERK activation also occurred in TLR2 transfectant HEK293 cells with 30kappaDa antigen stimulation. CONCLUSION: 30kappaDa antigen is one of the major mycobacterial antigens inducing cytokine production and MAP kinases phosphorylation in macrophages.
Blotting, Western ; Cytokines ; Dendritic Cells ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; HEK293 Cells ; Interleukin-12 ; Interleukin-6* ; Lipoproteins ; Macrophages* ; Phosphorylation ; Phosphotransferases ; Receptors, Pattern Recognition ; Tumor Necrosis Factor-alpha*

BACKGROUND: The expression of BRG1 associated factors (BAF) 155 and BAF 170 in response to IFN-gamma or TNF-alpha was studied in astrocytoma cell lines and primary astrocytes. BAFs are complexed with BRG1 and are also associated with activated glucocorticoid for glucocorticoid trans-activation. METHODS: IFN-gamma was pretreated for 18 hrs and cells were incubated with IL-1 or TNF-alpha for 72 hrs or 96 hrs with different concentrations of steroid. Cell death was measured by LDH assay. BAF expression was assayed by RT-PCR. RESULTS: IFN-gamma increased cell death by dexamethasone in LN215 cells but not in LN319 cells. The IFN-gamma increased the expression of BAF 155 and BAF 170 in adult astrocytes and LN215 cells, but IFN-gamma decreased the expression of BAF 155/170 in LN319 cells. The effect of IFN-gamma on the expression of BAF was not as clear in fetal astrocytes as it was in adult astrocytes. CONCLUSION: Our results suggest cytokines produced during immune reaction or immunotherapy may modulate steroid susceptibility of astrocytes and astrocytoma cells by influencing the expression of BAFs.
Adult ; Astrocytes* ; Astrocytoma ; Cell Death ; Cell Line ; Cytokines ; Dexamethasone ; Humans ; Immunotherapy ; Interleukin-1 ; Tumor Necrosis Factor-alpha

Silica nanoparticles, which are applicable in many industrial fields, have been reported to induce cellular changes such as cytotoxicity in various cells and fibrosis in lungs. Because the immune system is the primary targeting organ reacting to internalized exogenous nanoparticles, we tried to figure out the immunostimulatory effect of silica nanoparticles in macrophages using differently sized silica nanoparticles. Using U937 cells we assessed cytotoxicity by CCK-8 assay, ROS generation by CM-H2DCFDA, intracellular Ca++ levels by staining with Fluo4-AM and IL-8 production by ELISA. At non-toxic concentration, the intracellular Ca++ level has increased immediately after exposure to 15 nm particles, not to larger particles. ROS generation was detected significantly in response to 15 nm particles. However, all three different sizes of silica nanoparticles induced IL-8 production. 15 nm silica nanoparticles are more stimulatory than larger particles in cytotoxicity, intracellular Ca++ increase and ROS generation. But IL-8 production was induced to same levels with 50 or 100 nm particles. Therefore, IL-8 production induced by silica nanoparticles may be dependent on other mechanisms rather than intracellular Ca++ increase and ROS generation.
Enzyme-Linked Immunosorbent Assay ; Fibrosis ; Immune System ; Interleukin-8 ; Lung ; Macrophages ; Nanoparticles ; Silicon Dioxide ; Sincalide ; U937 Cells

165Dy Hydroxide Macroaggregates(165Dy HMA) has a short half life(2.3 hours) and a size range of 3-5µm that give the advantage of reduced leakage and a shorter hospital stay. This report will show the results of a prospective open study on the efficacy and safety of 165Dy HMA in 178 knees of 141 patients with chronic synovitis refractory to conventional antirheumatic therapy. The final global assessment was classified as good, fair or poor. Extra-articular leakage of 165Dy HMA was determined by the scintigraphic evaluation of liver, groin and knee joints. The optimum radiation dose was 250 mCi. The mean follow up periods were 32.4(14-112) weeks. Thirty seven percent of the knees showed good results, 48% fair results and 15% poor results. In the knees with stage I radiographic changes, 82% showed improvement including 32% of the patients with good results. In the knees with stage II radiographic changes, 90% showed improvement including 42% of the patients with good results. The mean period of improvement for the 158 knees that responded to treatment was 41.4(24-106) weeks. Leakage of radioactivity from the injected joint was minimal. Adverse reactions were rare(radiation burn : 4 cases, transient postinjection swelling : 14 cases). In conclusion, 165Dy HMA radiation synovectomy is a safe and useful therapy for chromic synovitis of the rheumatoid knees.
Arthritis, Rheumatoid ; Burns ; Follow-Up Studies ; Groin ; Humans ; Injections, Intra-Articular ; Joints ; Knee Joint ; Knee ; Length of Stay ; Liver ; Prospective Studies ; Radioactivity ; Synovitis

Since CKD-712 has been developed as an anti-inflammatory agent, we examined the effect of CKD-712 during TLR4 signaling. Using HEK293 cells expressing TLR4, CKD-712 was pre-treated 1 hr before LPS stimulation. Activation of NF-kappaB was assessed by promoter assay. The activation of ERK, JNK, p38, IRF3 and Akt was measured by western blotting. CKD-712 inhibited the NF-kappaB signaling triggered by LPS. The activation of ERK, JNK, p38 or IRF3 was not inhibited by CKD-712. On the contrary the activation of these molecules was augmented slightly. The activation of Akt with stimulation of LPS was also enhanced with CKD-712 pre-treatment at lower concentration, but was inhibited at higher concentration. We suggest that during TLR4 signaling CKD-712 inhibits NF-kappaB activation. However, CKD-712 augmented the activation of Akt as well as Map kinases. Therefore, we suggest that CKD-712 might have a role as an immunomodulator.
Blotting, Western ; HEK293 Cells ; NF-kappa B ; Phosphotransferases ; Tetrahydroisoquinolines