BACKGROUND: Leptin is a 16-KDa non-glycosylated peptide hormone synthesized almost exclusively by adipocytes. The well-known function of leptin is regulation of food intake and energy expenditure. Leptin also plays a regulatory role in immune and inflammatory process including cytokine production. The purpose of this study was to investigate the effect of leptin on the expression of several chemokine genes(RANTES, IL-8, MCP-1, IP-10, Mig, MIP-1alpha, MIP-1beta, and GRO-alpha) in THP-1 cells. MATERIALS AND METHODS: Total RNA of THP-1 cells were prepared by Trizol method, and then stimulated with the leptin(250 ng/microliter) or LPS(100 ng/microliter). We examined the expression patterns of various chemokine mRNAs in THP-1 cell lines by RT-PCR and Northern blot. RESULTS: Leptin did not induce the expression of chemokine mRNAs in THP-1 cells. The expression patterns of RANTES, IL-8, MCP-1, IP-10, and Mig mRNAs in THP-1 cells stimulated with leptin and LPS simultaneously was almost same to the patterns of LPS alone-induced chemokine mRNAs. RANTES mRNA expression was independent on the concentrations of leptin. Although leptin did not have strong effect on the expression of RANTES, IL-8, MCP-1, IP-10, Mig, MIP-1alpha, MIP-1beta, and GRO-alpha mRNAs in THP-1 cells, leptin could induce the expression of long isoform of leptin receptor(OB-RL) mRNA, and its expression was elevated in simultaneous stimulation of leptin and LPS. CONCLUSION: These data suggest that leptin is able to induce OB-RL in THP-1 cells, however, leptin has little effect on the expression of pro-inflammatory chemokine genes.
Adipocytes ; Blotting, Northern ; Cell Line ; Chemokine CCL3 ; Chemokine CCL4 ; Chemokine CCL5 ; Eating ; Energy Metabolism ; Interleukin-8 ; Leptin ; RNA ; RNA, Messenger

BACKGROUND AND OBJECTIVES: The number of eosinophil in nasal polyps has been reported to be strongly elevated when compared to non-affected nasal tissue, indicating an important role for eosinophils in the pathogenesis of nasal polyposis. The mechanisms determining selective eosinophilic tissue infiltration into diseased nasal mucosa as yet are specualtive. Panleukotactic factors also known to be present on nasal polyps cannot explain the type-selective tissue infiltration in eosinophilic or neutrophilic-featured diseases. Chemokines are known to have leukocyte subtype-selective chemotactic properties in vitro and thus are candidates explaining leukocytic characteristic tissue infiltration. The aim of this study was to investigate whether specific chemokines are associated with different forms of nasal polyps. This study was designed to demonstrate the expressions of various CC chemokines. MATERIALS AND METHODS: Nasal polyp from patients with systemic allergy (AP group, n=7) and negative allergic skin tests (NP group, n=10) were sampled. Expressions of RANTES, eotaxin, MCP-1, MIP-1alpha,beta were studies by RT-PCR and immunohistochemical studies. RESULTS: Expression and mean density of RANTES, MCP-1, MIP-1beta were significantly stronger in NP group than in AP group (p<0.05). However, those of eotaxin and MIP-1alpha were significantly stronger in AP group than in NP group (p<0.01). CONCLUSION: This results suggest that only selective chemokines could be involved to develop the pathologic conditions in different type of nasal polyp.
Chemokine CCL3 ; Chemokine CCL4 ; Chemokine CCL5 ; Chemokines ; Chemokines, CC* ; Eosinophils ; Humans ; Hypersensitivity ; Leukocytes ; Nasal Mucosa ; Nasal Polyps* ; Skin Tests

BACKGROUND AND OBJECTIVES: The cause of nasal polyp is unsure but inflammation is thought to be an important factor in the development of nasal poyps. CC chemokine is a powerful chemotactic cytokine for inflammatory cells. We designed this study to investigate whether specific CC chemokines are associated with different forms of nasal polyps and their changes according to antibiotic treatment. MATERIALS AND METHODS: Nasal polyp from patients with atopy (AP group, n=12) and without atopy (NP group, n=20) were sampled. Expressions of RANTES, eotaxin, MCP-2, MCP-3 and MIP-1alpha were studied by an immunohistochemical study. Specimens of non-allergic nasal turbinates were used as the control group from 14 patients who were operated for nasal blockage. All patients were divided into 2 groups. One group was treated with antibiotics for 10 days before operation. The other was non-treated. RESULTS: Between the NP and AP group, the ratio of stained cells except anti-MCP 2 monoclonal antibody in the AP group was more increased than that of the NP group. Among them, RANTES and eotaxin were increased significantly (p<0.05). There was a significant difference of the expression of 5 CC chemokines between the treated and non-treated groups (p<0.05). CONCLUSIONS: These results suggest that chemokines play an important role in the development of nasal polyps, and different kinds of chemokines can be involved according to the cause of nasal polyps and CC chemokines affected by antibiotic treatment.
Anti-Bacterial Agents ; Chemokine CCL3 ; Chemokine CCL5 ; Chemokines ; Chemokines, CC ; Humans ; Inflammation ; Nasal Obstruction ; Nasal Polyps* ; Turbinates

PURPOSE: Portal vein transfusion (PVT) has been known to induce immunosuppression or tolerance and Kupffer cell was identified to play an important role in the phenomenon. The purposes of this study were investigating PVT effect on gene regulation in Kupffer cells and subsequent change in serum cytokine. METHODS: For investigating the effect of PVT, Kupffer cells were isolated from the mice (BalbC) of six groups; 1 hour sham operation (S), 1 hour portal vein saline injection (PVS), 1 hour PVT, 24 hour S, 24 hour PVS, and 24 hour PVT groups. Each group was composed of 3 mice. Total RNAs isolated from Kupffer cells were subjected to RT-PCR differential display. The bands of 24 hour group showing increased expression was cloned for the sequencing analysis. RESULTS: Macrophage inflammatory protein 1 alpha (MIP-1alpha) was identified from the bands of increased expression. In PVT groups, increased expression of MIP-1alpha mRNA in Kupffer cells coincided with elevated serum level of MIP-1alpha. CONCLUSION: MIP-1alpha may be one of the important cytokines involved in PVT induced immunosuppression or tolerance.
Animals ; Chemokine CCL3* ; Clone Cells ; Cytokines ; Immunosuppression ; Kupffer Cells* ; Mice ; Portal Vein* ; RNA ; RNA, Messenger*

BACKGROUND: We tested the hypothesis that dengue haemorrhagic fever (DHF) is associated with a T(H)1-skewed immune response as opposed to dengue fever (DF). METHODS: We estimated intracellular (in T-cells) and serum levels of designate T(H)1/T(H)2 cytokines [interferon-gamma (IFN-gamma), interleukin-4 (IL-4), and tumor necrosis factor-alpha] and macrophage inflammatory protein-1alpha (MIP-1alpha) at admission, 48 h, and day 5 in 20 adults with dengue (DF=10, DHF=10) and 10 dengue-naive healthy controls. RESULTS: At admission, intracellular IFN-gamma/IL-4 ratio in CD4+ T-cells and proportion of MIP-1alpha-positive CD8+ T-cells were significantly higher in patients with DHF [7.21 (5.36~10.81) vs. 3.04 (1.75~4.02); p=0.011 and 6.2% (3.2~8.2%) vs. 2.4% (2.0~3.6%); p=0.023]. The latter showed a significant positive correlation with IFN-gamma/IL-4 ratio in CD4+ T-cells (Spearman's rho=0.64; p=0.003), percentage-change in haematocrit (rho=0.47; p=0.048), and serum alanine aminotransferase level (rho=0.61; p=0.009). CONCLUSION: We conclude that DHF is associated with a T(H)1-skewed immune response. Further, MIP-1alpha in CD8+ T-cells is an important immunologic correlate of disease severity in dengue.
Adult ; Alanine Transaminase ; Chemokine CCL3 ; Cytokines ; Dengue ; Fever ; Humans ; Interferon-gamma ; Interleukin-4 ; Macrophages ; Necrosis ; T-Lymphocytes

BACKGROUND/AIMS: To elucidate chemokine gene program in gastric mucosa infected with H. pylori, we quantified mRNA expression of CXC [growth-related oncogene alpha(GROalpha) and epithelial cell- derived neutrophil activating protein-78 (ENA-78)] and CC [macrophage inflammatory protein (MIP)- 1alpha and MIP-1beta] chemokines. METHODS: Forty-seven patients with dyspeptic symptoms undergoing diagnostic gastroduodenoscopy were enrolled. The presence of H. pylori was identified by histology and rapid urease test. RNA was extracted from gastric antral mucosa and quantitative RT-PCR was performed using synthetic standard RNA. RESULTS: GROalpha and ENA-78 mRNA transcripts in H. pylori-infected mucosa showed 14- and 15-fold increase, as compared with non-infected mucosa, respectively (p<0.01). GROalpha and ENA-78 transcripts tended to be correlated with the degree of neutrophil infiltration. ENA-78 transcripts of patients with gastric ulcer were significantly higher than those of the non-ulcer patients (p<0.05). In case of MIP-1alpha and MIP-1beta, statistical analysis for above parameters showed no significant difference in mRNA transcripts. After eradication of H. pylori, all of GROalpha, ENA-78, MIP-1alpha and MIP-1beta transcripts significantly decreased, as compared with pre-treatment levels. CONCLUSION: Upregulation of CXC chemokine genes may play a central role in gastric mucosal inflammation induced by H. pylori. Overexpression of ENA-78 gene may be important in the pathogenesis of gastric ulcer.
Chemokine CCL3 ; Chemokine CCL4 ; Chemokines ; Chemokines, CC* ; Gastric Mucosa* ; Helicobacter pylori* ; Helicobacter* ; Humans ; Inflammation ; Mucous Membrane ; Neutrophil Infiltration ; Neutrophils ; Oncogenes ; RNA ; RNA, Messenger ; Stomach Ulcer ; Up-Regulation ; Urease

Nystatin, a polyene antifungal antibiotic, is a cholesterol sequestering agent. The antifungal agent alters composition of the plasma membrane of eukaryotic cells, whereas its effects on cells are poorly investigated. In the current study, we investigated the question of whether nystatin was able to induce expression of macrophage inflammatory protein-1 (MIP-1). THP-1 cells rarely express MIP-1alpha and MIP-1beta, however, upon exposure to nystatin, significantly elevated expression of MIP-1alpha and MIP-1beta was observed in a dose-dependent fashion at the messenger and protein levels. Cellular factors activated by nystatin as well as involved in nystatin-induced expression of MIP-1 proteins were identified in order to understand the molecular mechanisms of action of the anti-fungal agent. Treatment with nystatin resulted in enhanced phosphorylation of Akt, ERK, p38 MAPK, and JNK. Abrogation or significant attenuation of nystatin-induced expression of MIP-1alpha and MIP-1beta was observed by treatment with Akt inhibitor IV, LY294002, and SP6001250. Inhibition of ERK or p38MAPK using U0126 and SB202190 did not lead to attenuation of MIP-1 expression. In addition, inhibitors of protein kinase C, such as GF109203X and Ro-318220, also attenuated expression of MIP-1. These results indicate that nystatin is able to activate multiple cellular kinases and, among them, Akt and JNK play primary roles in nystatin-induced expression of MIP-1 proteins.
Cell Membrane ; Chemokine CCL3 ; Chemokine CCL4 ; Cholesterol ; Eukaryotic Cells ; Macrophage Inflammatory Proteins* ; Macrophages* ; Nystatin* ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Phosphotransferases ; Protein Kinase C

The cytokines are the hormone-like proteins, which are produced in the mononuclear cells. They have many roles, such as immune mediators, cell differentiations, angiogenesis. The chemokines have chemotactic effects which control the host immune response. There were few reports about the cytokines associated with musculoskeletal tumors. From late 1980s, the cytokine studies of bone tumors such as osteosarcoma were started, but most studies for benign and malignant musculoskeletal tumors were left to be explored. To evaluate the characteristics of the cytokines in variable musculoskeletal tumors, tissues were obtained from the seven patients who visited the Yeungnam University hospital from February to July 2000. They were lipoma (1 case), parosteal osteoma (1 case), enchondroma (2 cases), pigmented villonodular synovitis (1 case), ganglion (1 case), and metastaic squamous cell carcinoma (1 case). The gene experession of the cytokines were analyzed by RNase protection assay (RPA) and reverse transcription-polymerase chain reaction (RT-PCR). The lipoma and parosteal osteoma expressed MIP-1beta, and IP-10 genes. The two enchondromas showed different results, one expressed all of MIP-1alpha, MIP-1beta and IP-10 genes but the other expressed none of above. The pigmented villonodular synovitis strongly expressed MIP-1alpha and IP-10 when compared with the other cases. The ganglion did not express all of the chemokines mentioned above. And the metastatic squamous cell carcinoma expressed all of the chemokines and especially IP-10 was highly expressed. Even though this study has only a few cases, these results provide a basis for the cytokine mediating network study in musculoskeletal tumors.
Carcinoma, Squamous Cell ; Cell Differentiation ; Chemokine CCL3 ; Chemokine CCL4 ; Chemokines ; Chondroma ; Cytokines ; Ganglion Cysts ; Humans ; Lipoma ; Negotiating ; Osteoma ; Osteosarcoma ; Ribonucleases ; Synovitis, Pigmented Villonodular

BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) belongs to C-C subfamily of chemokines, which stimulates the migration of monocytes. MCP-1 exerts various effects on the monocytes, including the induction of integrin and tissue factor, and synthesis of proinflammatory cytokines and arachidonic acid. In this study, we measured the MCP-1 levels in patients with Behcet's disease and evaluated the associations between the levels of MCP-1 and the level of other chemokines and various clinical features of Behcet's disease. METHODS: Serum samples were obtained from 67 patients with Behcet's disease and 30 healthy controls. Simultaneously, whole blood was isolated from patients (n=25) with Behcet's disease and healthy controls (n=11) and cultured in 24 well plates for 48 hours in the absence or presence of lipopolysaccharide (LPS) 5 microgram/mL, phytohaemagglutinin (PHA) 5 microgram/mL, phorbol 12-myristate 13-acetate (PMA) 50 ng/mL + ionomycin 5 microgram/mL. The MCP-1 concentrations were measured in the sera and culture supernatants by enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of serum MCP-1 were 2.5 times higher in patients with Behcet's disease than healthy controls. The patients with Behcet's disease had also higher levels of MCP-1 in the culture supernatants of whole blood cells, stimulated with LPS, but not with either PHA or PMA plus ionomycin, compared to healthy controls. Serum MCP-1 levels (n=67) were strongly correlated with serum RANTES, MIP-1alpha, IL-8 levels in Behcet's disease. In addition, the production of MCP-1 by whole blood culture from Behcet's disease patients (n=25) were also correlated well with those of RANTES, MIP-1alpha, and IL-8, when stimulated with LPS. However, MCP-1 levels in the sera and culture supernatants did not show any association with various clinical features of Behcet's disease including oral ulcer, genital ulcer, erythema nodosum, arthritis, uveitis, intestinal involvement, central nervous system involvement, and vascular thrombosis. CONCLUSION: In the sera and culture supernatants of whole blood stimulated with LPS, MCP-1 levels were higher in patients with Behcet's disease than controls and correlated well with RANTES, MIP-1alpha, IL-8 levels. These results suggest that the activation and migration of monocytes triggered by the increased production of MCP-1 may play a role in the pathogenesis of Behcet's disease.
Arachidonic Acid ; Arthritis ; Blood Cells ; Central Nervous System ; Chemokine CCL2* ; Chemokine CCL3 ; Chemokine CCL5 ; Chemokines ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Erythema Nodosum ; Humans ; Interleukin-8 ; Ionomycin ; Monocytes* ; Oral Ulcer ; Thromboplastin ; Thrombosis ; Ulcer ; Uveitis

Osteoblasts regulate osteoclastogenesis by production of various cytokines. Aggregatibacter(A) actinomycetemcomitans is one of periodontopathogens which invades gingival tissue. Therefore, clarifying the effect of alive A. actinomycetemcomitans on osteoblasts is important to understand the mechanism of alveolar bone resorption in periodontitis. We investigated induction of osteoclastogenesis- inducing cytokines, adherence, and invasion by A. actinomycetemcomitans in osteoblasts. Osteoblasts were isolated from mouse calvaria and expression of cytokines was determined by RT-PCR. When the ratio of the number of A. actinomycetemcomtians to the number of osteoblasts was 10:1, 50:1 and 100:1, RANKL mRNA expression was increased. A. actinomycetemcomitans also increased expression of macrophage inflammatory protein (MIP)-1alpha, interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha. A. actinomycetemcomitans attached to and invaded osteoblasts at ratio of 1000:1. These results suggest that A. actinomycetemcomitans increases osteoclastogenesis-inducing ability of osteoblasts by stimulating the expression of RANKL, MIP-1alpha, IL-1beta, and TNF-alpha and that invasion of A. actinomycetemcomitans provides a means by which the bacteria escape from immune system and antibiotic therapy.
Aggregatibacter actinomycetemcomitans* ; Aggregatibacter* ; Animals ; Bacteria ; Bone Resorption ; Chemokine CCL3 ; Cytokines* ; Immune System ; Interleukins ; Macrophages ; Mice ; Osteoblasts* ; Periodontitis ; RNA, Messenger ; Skull ; Tumor Necrosis Factor-alpha ; United Nations