PURPOSE: The pathologic mechanisms of central nervous system(CNS) injuries in human meningitis are not yet completely understood. Recent studies indicate that the host inflammatory responses are as important in brain damage as the infecting organisms and toxins. There have been some reports on the relationship of nitric oxide(NO), macrophage inflammatory protein-1 alpha (MIP-1 alpha ), and lactoferrin in bacterial meningitis, but few reports in aseptic meningitis. Thus, we investigated the concentrations of NO, MIP-1 alpha and lactoferrin in cerebrospinal fluid(CSF) and serum of patients with aseptic meningitis and control subjects and evaluated their relationship with other parameters of meningitis. METHODS: CSF and blood were obtained from 25 subjects with aseptic meningitis and 15 control subjects. After centrifugation, supernatants were stored at -70degrees C and we assayed the concentrations of NO, MIP-1 alpha and lactoferrin with the ELISA method. There were no patients with neurologic sequelae after being recovered from aseptic meningitis. RESULTS: Concentrations of CSF and serum NO, MIP-1 alpha were not increased in aseptic meningitis subjects compared to control subjects. Concentration of CSF lactoferrin was significantly elevated in patients with aseptic meningitis and concentration of serum lactoferrin was significantly decreased in patients with aseptic meningitis compared with those in control subjects(P<0.05). CSF lactoferrin level was positively correlated with CSF WBC counts(rs=0.449, P=0.007), especially with neutrophil counts(rs=0.574, P<0.001) and CSF protein level(rs=0.508, P=0.002). CONCLUSION: Lactoferrin plays an important role in aseptic meningitis and may be released from neutrophils recruited from blood to the CSF through breakdown of blood-brain barrier. NO and MIP-1 alpha may not be important factors in the pathogenesis of aseptic meningitis without neurologic sequelae.
Blood-Brain Barrier ; Brain ; Centrifugation ; Enzyme-Linked Immunosorbent Assay ; Humans ; Lactoferrin* ; Macrophage Inflammatory Proteins* ; Macrophages* ; Meningitis ; Meningitis, Aseptic* ; Meningitis, Bacterial ; Neutrophils ; Nitric Oxide*

BACKGROUND: The induction of production and production inhibition of alpha1-antichymotrypsin (ACT), IL-1 alpha, IL-1 alpha receptor and macrophage inflammatory protein-1 (MIP-1) receptor in A beta1-42 (A beta)-stimulated U373MG cell, the human astrocytoma cell line, have never been reported. METHODS: U373MG cells (1 x 10(6) cells in RPMI-1640 media) were incubated for overnight after administration of a single dose of 20 micro M of A beta or 0.5 ng/ml of TNF alpha or both. Actinomycin D (2.5 micro M) or cycloheximide (2.5 micro M) was also added to the cell suspension. Messenger RNA expression of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 receptor was measured by RT-PCR. Western blot was done and nitrocellulose paper was stained with anti-ACT and anti-GFAP antibody. NF kappa B activation after treatment of A beta in U373MG cells was detected by electrophoretic mobility-shift assay. RESULTS: A beta and TNF alpha both increased production of ACT in a dose-dependent manner. TNF alpha enhanced A beta-induced mRNA had increases of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor. Activated NF kappa B was demonstrated in the A beta, TNF alpha-stimulated U373MG cells. Actinomycin suppresses mRNA level of ACT and IL-1 alpha receptor but cycloheximide inhibits the expression of ACT, IL-1 alpha and MIP-1 alpha receptor. CONCLUSIONS: TNF alpha increases synthesis of ACT, IL-1 alpha, IL-1 alpha receptor and MIP-1 alpha receptor in A beta-stimulated astrocyte, which, as a result, may contribute to the neuroinflammation of Alzheimer's disease.
Alzheimer Disease ; Amyloid beta-Peptides* ; Astrocytes ; Astrocytoma* ; Blotting, Western ; Cell Line* ; Collodion ; Cycloheximide ; Dactinomycin ; Humans* ; Interleukin-1 ; Interleukin-1alpha ; Macrophage Inflammatory Proteins ; NF-kappa B ; RNA, Messenger

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

Animals ; Chemokine CCL20 ; Chemokine CXCL10 ; Chemokines, CC ; biosynthesis ; Chemokines, CXC ; biosynthesis ; Liver ; metabolism ; Liver Transplantation ; Macrophage Inflammatory Proteins ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transplantation, Homologous

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

BACKGROUND: Chemokines are molecules with chemotatic activities on selective leukocyte populations and are sub-grouped into alpha-chemokine acting primarily on PMNL (polymorphonuclear leukocyte) and beta-chemokines attracting mainly lymphocytes and monocytes. We conducted a prospective study to investigate the serum levels of interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1, and macrophage inflammatory protein (MIP)-1 alpha in patients with acute ischemic stroke and carotid atherosclerosis. METHODS: Serum was sampled from patients with acute ischemic stroke (<24hrs), with persistent ischemic neurological deficits associated with atherosclerosis (>1 month), and from normal subjects without a history of vascular disease. Concentrations of chemokines were measured by enzyme linked immunosorbent assay ( ELISA ). RESULTS: Compared with carotid atherosclerotic patients and control subjects, the serum levels of IL-8 were significantly elevated in those with acute ischemic stroke. The serum levels of MCP-1 in patients with large artery disease were higher than those in patients with small vessel disease and cardioembolism. CONCLUSIONS: This study suggested that IL-8 can be involved in acute ischemic stroke and MCP-1 plays a role in the pathogenesis of atherosclerosis.
Arteries ; Atherosclerosis* ; Carotid Artery Diseases ; Chemokine CCL2* ; Chemokines ; Chemokines, CC ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-8* ; Interleukins ; Leukocytes ; Lymphocytes ; Macrophage Inflammatory Proteins* ; Macrophages* ; Monocytes* ; Prospective Studies ; Stroke* ; Vascular Diseases

This study was aimed to explore the expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 in bcr/abl fusion gene positive CML cells, and to study the effects of P210(bcr/abl) fusion protein tyrosine kinase on expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNAs in chronic myeloid leukemia cells. The expression levels of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNA were detected by semi-quantitative RT-PCR in bcr/abl negative cells, bcr/abl positive cells, and P210(bcr/abl)-Rb-C-Box positive cells. The results showed that MIP-1alpha and CCR-1 mRNAs were expressed in bcr/abl negative cells, but not in positive cells. Both MCP-1 and CCR-2 mRNA cannot be detected in both bcr/abl positive and negative cells. After inhibiting P210(bcr/abl) tyrosine kinase activity by Rb-C-Box, expressions of MIP-1alpha and CCR-1 mRNAs were restored to normal (similar to P210(bcr/abl) negative cells). It is concluded that P210(bcr/abl) fusion protein inhibits the expression of MIP-1alpha and CCR-1 in chronic myeloid leukemia cells, but does not inhibit MCP-1 and CCR-2 mRNA expressions in these leukemia cells.
Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Receptors, CCR1 ; Receptors, CCR2 ; Receptors, Chemokine ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVE: To explore the microvessel counts and the expressions of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1 ( MIP-1) alpha mRNA in the pathological scar tissues. METHODS: Immunohistochemical method of avidin-biotin complex was used for microvessel counts on the routinely formalin-fixed and paraffin-embedded sections of specimens of hypertrophic scars, keloids, normal skin, and surgical scar, and in situ hybridization for the expressions of IL-8, MCP-1, MIP-1alpha mRNA. RESULTS: The microvessel counts as well as the positive rates and the scorings of IL-8, MCP-1, and MIP-1alpha mRNA were significantly higher in pathological scars than those in the normal skin and surgical scar (all P < 0.05). The microvessel counts were significantly higher in the positive cases of IL-8, MCP-1 and MIP-1alpha mRNA than those in the negative ones (P < 0.05). The close positive correlations were found among the microvessel counts and the expressive scorings of 3 factors (P < 0.05). The close positive correlations were also found among the expressive scorings of IL-8, MCP-1, and MIP-1alpha mRNA in pathological scars. Microvessel counts were significantly higher in hypertrophic scars with the course less than 1 year than those with the course more than 1 year. CONCLUSION IL-8, MCP-1 and MIP-1alpha play important roles in promoting the neovascularization of pathological scars.
Adolescent ; Adult ; Burns ; complications ; Capillaries ; metabolism ; Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Cicatrix ; etiology ; metabolism ; Female ; Humans ; Interleukin-8 ; biosynthesis ; genetics ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Skin ; blood supply

OBJECTIVE: To detect the expressions of chemokines interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1 (MIP-1) mRNA in non-small cell lung cancer (NSCLC) tissues, to analyze their relationship with microvessel counts (MVC), and their significance in clinic pathologic features of NSCLC. METHODS: In situ hybridization was used to measure the expressions of chemokine IL-8, MCP-1, and MIP-1 mRNA in 40 NSCLC tissues and 10 normal pulmonary tissues, and immunohistochemical staining was carried out to measure the MVC in the above tissues. RESULTS: The positive ratios of IL-8, MCP-1, and MIP-1 mRNA in the 40 NSCLC tissues were apparently higher than those in the 10 normal contrast tissues and the difference was statistically significant. The numbers varied accordingly with the different clinic pathologic features of NSCLC, showing that Group T(3) > Group T(2) or Group T(1), Group III stage> Group II stage> Group I stage Group lymph node and remote transferred > Group non-transferred, and Group of survival time no more than 3 years > Group of survival time more than 3 years. The positive expressions among IL-8, MCP-1,and MIP-1 mRNA and between these and the MVC all had mutually positive correlation. CONCLUSION Chemokine IL-8, MIP-1, and MIP-1 in NSCLC tissues might cooperate with one another to promote the tumor angiogenesis and affect the progression, metastasis and prognosis of the tumor.
Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; blood supply ; metabolism ; Chemokine CCL2 ; metabolism ; Female ; Humans ; Interleukin-8 ; metabolism ; Lung Neoplasms ; blood supply ; metabolism ; Macrophage Inflammatory Proteins ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; Prognosis