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: 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

PURPOSE: To investigate the expression of CCL4/MIP-1beta in tears of patients with dry eye syndrome and its correlation with the tear surface parameters. METHODS: Forty patients with dry eye syndrome (seven Meibomian gland dysfunction and 33 non-Meibomian gland dysfunction patients) and 25 control subjects were recruited. The concentrations of CCL4 in tears were measured using an enzyme-linked immunosorbent assay. The correlations between CCL4 levels and tear surface parameters including tear film break-up time (BUT), basal tear secretion (BST), tear clearance test (TCR), corneal sensation, keratoepitheliopathy (KEP) and conjunctival goblet cell density (GCD) were analyzed in patients with dry eye syndrome. RESULTS: The mean levels of MIP-1beta were 4.3+/-5.2 pg/ml in patients with dry eye syndrome and 81.6+/-294.3 pg/ml in control subjects (p=0.03). The mean levels in non-Meibomian gland disease patients (90.4+/-313.9 pg/ml) were significantly higher than those of Meibomian gland dysfunction patients (20.3+/-23.2 pg/ml, p=0.02). CCL4 levels correlated significantly with BST, TCR, KEP and GCD (p<0.05). CONCLUSIONS: The level of MIP-1beta is elevated in tears of patients with dry eye syndrome and correlates with disease severity and tear surface parameters.
Chemokine CCL4 ; Dry Eye Syndromes ; Enzyme-Linked Immunosorbent Assay ; Eye ; Goblet Cells ; Humans ; Meibomian Glands ; Sensation ; Tears

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

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

PURPOSE: To better understand the extent to which chemokines participate in the mucosal inflammatory response in patients with ulcerative colitis (UC), we assessed the expression of an array of chemokines in the colonic mucosa of UC patients. METHODS: Colonic mucosal biopsy specimens were obtained from 15 patients with UC and 12 normal controls. Messenger RNA (mRNA) levels for 10 chemokines were quantitated by reverse-transcription PCR using synthetic standard RNAs. The biopsy specimens were also cultured, and secreted chemokines in culture supernatants were assayed by ELISA. RESULTS: The mRNA expression of C-X-C (IL-8, GROalpha, GRObeta, GROgamma, ENA-78, and IP-10) and C-C (MCP-1, MIP-1beta, and RANTES), but not C (lymphotactin) chemokines was significantly higher in the affected mucosa of UC patients than in the unaffected mucosa of UC patients or in the normal mucosa of normal controls. The degree of increased expression was more prominent in the C-X-C than in the C-C chemokines. Further, the secretion of IL-8, GROalpha, ENA-78, and MCP-1 was higher in UC patients than in normal controls. Secretions of MIP-1beta and RANTES also showed a trend toward an increase in UC, but it did not reach statistical significance. CONCLUSION: The increased expression of a variety of chemokines in UC suggest that chemokines may play an important role in the immunopathogenesis of UC.
Biopsy ; Chemokine CCL4 ; Chemokine CCL5 ; Chemokines* ; Chemokines, CC ; Colitis, Ulcerative* ; Colon* ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-8 ; Mucous Membrane* ; Polymerase Chain Reaction ; RNA ; RNA, Messenger ; Ulcer*

UNLABELLEDBACKGROWND: Macrophage inflammatory protein-1α (MIP-l α/CCL3) belongs to the C-C chemokine family (CCL3), which can be secreted by macrophages, other types of hematopoietic cells and bone marrow stromal cells. Higher levels of MIP-1α were found to be associated with several kinds of hematologic malignancies, including multiple myeloma (MM), chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML). Moreover, MIP-1α has been reported to be an adverse prognostic factor for CLL. However, the impact of MIP-1α on acute myeloid leukemia (AML) has been poorly investigated.

OBJECTIVETo investigate the influence of MIP-1α on proliferction of AML cells.

METHODSUsing MLL-AF9 induced AML mouse model, the expression of MIP-1α was measured by real time quantitative RT-PCR. AML cell proliferation was examined by cell counting and colony forming assay (CFC). The influence of blocking the MIP-1α action on the growth and pathogenic ability of AML cells was explored by using the small molecule antagonist for interfering interaction of MIP-1α with its receptor CCR1.

RESULTSThe MIP-1α could promote the proliferation and colony formation of AML cells, the blocking MIP-1a could inhibit the growth of AML cells and delay onset of AML.

CONCLUSIONThe MIP-1a promotes the occurence and progression of AML, therefore blocking the MIP-1α signal pathway may be served as a strategy to inhibit the growth of AML cells, and MIP-1α can be a potential target for treatment of AML.

Animals ; Cell Line, Tumor ; Cell Proliferation ; Chemokine CCL3 ; Chemokine CCL4 ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Acute ; Macrophage Inflammatory Proteins ; Mice ; Multiple Myeloma ; Receptors, CCR1

OBJECTIVETo observe the in vivo antitumor activity of murine liver tumor vaccine expressing MIP-1alpha mediated by recombinant adenoviral vector.

METHODSThe infection efficacy was measured by GFP expression 48 hours after infection of Hepa1-6, and the number of cells was counted daily for 14 days. 5 x 10(6) modified Hepa1-6 cells were inoculated subcutaneously to C57BL/6 mice and the tumor-free animals were rechallenged by 2 x 10(6) wild-type Hepa1-6 cells or syngenic EL4 cells four weeks later. The tumor volume was measured twice a week.

RESULTSAdenoviral vectors could efficiently infect Hepa1-6 cells in vitro, and the in vitro growth rate of AdmMIP-1alpha modified Hepa1-6 cells was not affected; however the in vivo tumorigenicity was significantly decreased, compared with that of control vector modified Hepa1-6. Rechallenge of the tumor-free mice four weeks after administration of AdmMIP-1alpha with the parental Hepa1-6 cells resulted in significant inhibition of tumor growth, but there was no significant difference when rechallenged with EL4.

CONCLUSIONSThe liver cancer cells expressing mMIP-1alpha mediated by recombinant adenoviral vector decrease tumorigenicity and elicit specific immunological protection, and could be used as an effective liver tumor vaccine.

Adenoviridae ; genetics ; Animals ; Cancer Vaccines ; immunology ; Chemokine CCL3 ; Chemokine CCL4 ; Genetic Therapy ; Liver Neoplasms, Experimental ; therapy ; Macrophage Inflammatory Proteins ; genetics ; Mice ; Mice, Inbred C57BL ; Vaccines, Synthetic ; immunology