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1.
Article in English | WPRIM | ID: wpr-223789

ABSTRACT

Eupatilin is the main active component of DA-9601, an extract from Artemisia. Recently, eupatilin was reported to have anti-inflammatory properties. We investigated the anti-arthritic effect of eupatilin in a murine arthritis model and human rheumatoid synoviocytes. DA-9601 was injected into collagen-induced arthritis (CIA) mice. Arthritis score was regularly evaluated. Mouse monocytes were differentiated into osteoclasts when eupatilin was added simultaneously. Osteoclasts were stained with tartrate-resistant acid phosphatase and then manually counted. Rheumatoid synoviocytes were stimulated with TNF-alpha and then treated with eupatilin, and the levels of IL-6 and IL-1beta mRNA expression in synoviocytes were measured by RT-PCR. Intraperitoneal injection of DA-9601 reduced arthritis scores in CIA mice. TNF-alpha treatment of synoviocytes increased the expression of IL-6 and IL-1beta mRNAs, which was inhibited by eupatilin. Eupatilin decreased the number of osteoclasts in a concentration dependent manner. These findings, showing that eupatilin and DA-9601 inhibited the expression of inflammatory cytokines and the differentiation of osteoclasts, suggest that eupatilin and DA-9601 is a candidate anti-inflammatory agent.


Subject(s)
Animals , Anti-Inflammatory Agents/pharmacology , Arthritis, Experimental/chemically induced , Arthritis, Rheumatoid/drug therapy , Cell Differentiation/drug effects , Cells, Cultured , Collagen Type II , Cytokines/biosynthesis , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Female , Flavonoids/pharmacology , Humans , Inflammation/drug therapy , Interleukin-1beta/genetics , Interleukin-6/genetics , Lymph Nodes/cytology , Mice , Mice, Inbred DBA , Monocytes/cytology , Osteoclasts/cytology , Plant Extracts/pharmacology , RNA, Messenger/biosynthesis , Synovial Membrane/cytology , T-Lymphocytes, Regulatory/cytology , Tumor Necrosis Factor-alpha/pharmacology
2.
Braz. dent. j ; 25(2): 96-103, Mar-Apr/2014. graf
Article in English | LILACS | ID: lil-719224

ABSTRACT

Due to the critical role of monocytes/macrophages (Mϕ) in bone healing, this study evaluated the effects of bio-anodized, acid-etched, and machined titanium surfaces (Ti) on Mϕ behavior. Cells were separated from whole human blood from 10 patients, plated on Ti or polystyrene (control) surfaces, and cultured for 72 h. At 24, 48 and 72 h, cell viability, levels of IL1β, IL10, TNFα, TGFβ1 inflammatory mediators, and nitric oxide (NO) release were analyzed by mitochondrial colorimetric assay (MTT assay) and immunoenzymatic assays, respectively. Real-time PCR was used to verify the expression of TNFα and IL10 at 72 h. The data were subjected to a Kruskal-Wallis analysis. IL1β, TNFα and TGFβ1 release were not significantly different between the Ti surfaces (p>0.05). The presence of NO and IL10 was not detected in the samples. Cell viability did not differ between the samples cultivated on Ti and those cultivated on control surfaces, except at 24 h (p=0.0033). With respect to the mediators evaluated, the surface characteristics did not induce a typical Th1 or Th2 cytokine profile, although the cell morphology and topography were influenced by the Ti surface during the initial period.


Devido ao papel crítico dos monócitos / macrófagos (Mϕ) na cicatrização óssea, este estudo avaliou os efeitos de superficies de titânio (Ti) bio-anodizada, ataque ácido e usinada sobre o comportamento Mϕ. As células foram separadas a partir de sangue humano de 10 pacientes, plaqueadas em Ti ou superfícies de poliestireno (controle), e cultivadas durante 72 h. Às 24, 48 e 72 h, a viabilidade celular e IL1β, IL10, TNFα, TGFβ1 e liberação de óxido nítrico foram analisados por ensaio colorimétrico mitocondrial (MTT) e ensaios imunoenzimáticos, respectivamente. PCR em tempo real foi utilizado para verificar a expressão de TNFα e IL-10 às 72 h. Os dados foram submetidos a uma análise de Kruskal-Wallis. IL1β autorização, TNFα e TGFβ1, não foram significativamente diferentes entre as superfícies de Ti (p>0,05). A presença de NO e de IL-10 não foi detectada nas amostras. A viabilidade celular não diferiu entre as amostras cultivadas em Ti e aquelas cultivadas em superfícies de controle, exceto às 24 h (p=0,0033). No que diz respeito aos mediadores avaliados, as características da superfície, não induziu resposta típica de citocinas Th1 ou Th2, embora a morfologia da célula e a topografia foram influenciadas pela superfície de Ti, durante o período inicial.


Subject(s)
Humans , Macrophages/cytology , Monocytes/cytology , Titanium/chemistry , Cytokines/metabolism , Macrophages/metabolism , Monocytes/metabolism , Surface Properties
3.
Mem. Inst. Oswaldo Cruz ; 109(1): 70-79, 02/2014. graf
Article in English | LILACS | ID: lil-703644

ABSTRACT

Dendritic cells (DCs) are antigen (Ag)-presenting cells that activate and stimulate effective immune responses by T cells, but can also act as negative regulators of these responses and thus play important roles in immune regulation. Pro-angiogenic vascular endothelial growth factor (VEGF) has been shown to cause defective DC differentiation and maturation. Previous studies have demonstrated that the addition of VEGF to DC cultures renders these cells weak stimulators of Ag-specific T cells due to the inhibitory effects mediated by VEGF receptor 1 (VEGFR1) and/or VEGFR2 signalling. As the enzyme indoleamine 2,3-dioxygenase (IDO) is recognised as an important negative regulator of immune responses, this study aimed to investigate whether VEGF affects the expression of IDO by DCs and whether VEGF-matured DCs acquire a suppressor phenotype. Our results are the first to demonstrate that VEGF increases the expression and activity of IDO in DCs, which has a suppressive effect on Ag-specific and mitogen-stimulated lymphocyte proliferation. These mechanisms have broad implications for the study of immunological responses and tolerance under conditions as diverse as cancer, graft rejection and autoimmunity.


Subject(s)
Humans , Cell Proliferation/physiology , Dendritic Cells/drug effects , /metabolism , Lymphocytes/physiology , Vascular Endothelial Growth Factor A/pharmacology , Apoptosis , Antigens, Surface/biosynthesis , Cell Culture Techniques , Cells, Cultured , Cell Differentiation/physiology , Dendritic Cells/metabolism , Dendritic Cells/ultrastructure , Immune Tolerance/physiology , /genetics , Leukocytes, Mononuclear/physiology , Monocytes/cytology , Monocytes/ultrastructure , Necrosis , Real-Time Polymerase Chain Reaction , Signal Transduction/immunology
4.
Article in English | WPRIM | ID: wpr-158566

ABSTRACT

BACKGROUND: Numerous studies tried to find new markers that after hematopoietic stem cell transplantation predict engraftment earlier than the conventional marker, absolute neutrophil count (ANC >500/microL). Early engraftment prediction can be achieved by a marker that reflects the release of neutrophils and monocytes into the leukopenic peripheral blood. METHODS: We analyzed blood cell parameters, including cell population data such as volume, conductivity, and light scatter in 77 patients who underwent HSCT (allogeneic, n=63; autologous, n=11) to detect possible markers. RESULTS: We identified 2 early engraftment markers of neutrophils (NEUTRO) and monocytes (MONO); a pair of mean-volume-neutrophils (MNV) and mean-conductivity-neutrophils (MNC) for NEUTRO; and a pair of mean-volume-monocytes (MMV) and mean-conductivity-monocytes (MMC) for MONO. The new markers showed distinct patterns for early engraftment wherein 1) on the engraftment day, MNV peaked as MNC notched simultaneously for every case, and 2) MMV peaked as MMC notched simultaneously in most cases. Engraftment was predicted 3.8+/-2.7 days earlier than by ANC in 74 successful engraftment cases by using NEUTRO and/or MONO: 1) 72 cases (97.3%), in which NEUTRO and/or MONO predicted earlier engraftment than ANC, 2) 1 case, in which the 3 markers predicted engraftment on the same day, and 3) 1 case, in which NEUTRO predicted engraftment on the same day as ANC and MONO failed to predict engraftment. CONCLUSIONS: By analyzing the data from daily complete blood counts, engraftment can be predicted approximately 4 days earlier than ANC >500/microL using NEUTRO as a base marker and MONO as a supplementary marker.


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells/cytology , Humans , Infant , Leukocyte Count , Male , Middle Aged , Monocytes/cytology , Neutrophils/cytology , Time Factors , Transplantation, Autologous , Transplantation, Homologous , Young Adult
5.
Article in Korean | WPRIM | ID: wpr-150508

ABSTRACT

PURPOSE: This study was done to evaluate the effects of psychosocial interventions on cortisol and immune response in adult patients with cancer. METHODS: MEDLINE via PubMed, Cochrane Library CENTRAL, EMBASE, CINAHL and domestic electronic databases were searched. Twenty controlled trials (11 randomized and 9 non-randomized trials) met the inclusion criteria with a total of 862 participants. Methodological quality was assessed using the Cochrane's Risk of Bias for randomized studies and the Risk of Bias Assessment tool for non randomized studies. Data were analyzed using the RevMan 5.2.11 program of Cochrane library. RESULTS: Overall, study quality was moderate to high. The weighted average effect size across studies was -0.32 (95% CI [-0.56, -0.07], p=.010, I2=45%) for cortisol concentration, -0.62 (95%CI [-0.96,-0.29], p<.001, I2=0%) for T lymphocyte (CD3) and -0.45 (95%CI [-0.74, -0.16], p=.003, I2=0%) for Th lymphocyte (CD4) numbers. Psychosocial interventions were not effective for Tc lymphocyte (CD4), NK cell, monocyte, and cytokine response. CONCLUSION: Although these results provide only small evidence of successful immune modulation, they support the conclusion that psychosocial interventions can assist cancer patients in reducing emotional distress and improving immune response.


Subject(s)
CD4-Positive T-Lymphocytes/cytology , Cytokines/metabolism , Databases, Factual , Humans , Hydrocortisone/analysis , Killer Cells, Natural/cytology , Monocytes/cytology , Neoplasms/metabolism , Psychotherapy , T-Lymphocytes/cytology
6.
Braz. j. infect. dis ; 16(1): 19-26, Jan.-Feb. 2012. ilus
Article in English | LILACS | ID: lil-614545

ABSTRACT

OBJECTIVE: Epstein-Barr virus (EBV) is a ubiquitous human γ-herpes virus, which can adapt and evade host immune defense. Dendritic cells (DCs) play a pivotal role in the initiation and maintenance of immune responses. This study investigated the effects of EBV on cord blood monocytes derived DCs (CBDC). METHODS: Monocytes were isolated from cord blood and cultured in medium containing recombinant IL-4 and GM-CSF to induce DCs development. B95-8 supernatant was added in monocytes culture medium for EBV infection at day 0. Phenotypic characterization of DCs, apoptotic cells, and mitochondrial membrane potential (MMP) were detected by flow cytometry. The morphology was observed by Hoechst 33258 staining and TUNEL staining, the expression of X-linked inhibitor of apoptosis protein (XIAP) was detected by Western blotting assay and caspase 3, 8 and 9 activity was measured. RESULTS: Phenotypic characterization of DCs was changed in EBV-treated group. Chromatin condensation and DNA fragmentation were observed in EBV induced CBDC apoptosis. In addition, caspase 3, caspase 8, and caspase 9 activation were enhanced in the EBV-treated group. This was accompanied by the loss of MMP. Furthermore, XIAP expression was down-regulated in the EBV-treated group and compared to mock-infected group. CONCLUSION: These results suggested that EBV could inhibit CBDC phenotypic differentiation, and induce CBDC apoptosis in caspase-dependent manner with involvement of the mitochondrial pathway. This might help EBV to evade host immune responses to establish persistent infection.


Subject(s)
Humans , Apoptosis/physiology , Cytopathogenic Effect, Viral/physiology , Dendritic Cells/pathology , Fetal Blood/cytology , /physiology , Monocytes/pathology , Blotting, Western , Cell Differentiation , Caspases/immunology , Dendritic Cells/virology , Flow Cytometry , /immunology , /immunology , Monocytes/cytology , Monocytes/virology , Phenotype , X-Linked Inhibitor of Apoptosis Protein/immunology
7.
Article in English | WPRIM | ID: wpr-202339

ABSTRACT

Flow mediated brachial dilatation (FMD) and carotid intima-media thickness (IMT) have been a surrogate for early atherosclerosis. Slow coronary flow in a normal coronary angiogram is not a rare condition, but its pathogenesis remains unclear. A total of 85 patients with angina were evaluated of their brachial artery FMD, carotid IMT and conventional coronary angiography. Coronary flow was quantified using the corrected thrombosis in myocardial infarction (TIMI) frame count method. Group I was a control with normal coronary angiography (n = 41, 56.1 +/- 8.0 yr) and group II was no significant coronary stenosis with slow flow (n = 44, 56.3 +/- 10.0 yr). Diabetes was rare but dyslipidemia and family history were frequent in group II. Heart rate was higher in group II than in group I. White blood cells, especially monocytes and homocysteine were higher in group II. The FMD was significantly lower in group II than in group I. Elevated heart rate, dyslipidemia and low FMD were independently related with slow coronary flow in regression analysis. Therefore, endothelial dysfunction may be an earlier vascular phenomenon than increased carotid IMT in the patients with slow coronary flow.


Subject(s)
Aged , Angina, Unstable/complications , Brachial Artery/physiopathology , Carotid Intima-Media Thickness , Coronary Angiography , Coronary Circulation/physiology , Dyslipidemias/complications , Endothelium, Vascular/physiopathology , Female , Heart Rate , Homocysteine/metabolism , Humans , Leukocyte Count , Male , Middle Aged , Monocytes/cytology , ROC Curve , Regression Analysis , Risk Factors
8.
Article in English | WPRIM | ID: wpr-210394

ABSTRACT

A variety of benzylidenethiazole analogs have been demonstrated to inhibit 5-lipoxygenase (5-LOX). Here we report the anti-atherogenic potential of 5-(4-hydroxy-2,3,5-trimethylbenzylidene) thiazolidin-2,4-dione (HMB-TZD), a benzylidenethiazole analog, and its potential mechanism of action in LDL receptor-deficient (Ldlr-/-) mice. HMB-TZD Treatment reduced leukotriene B4 (LTB4) production significantly in RAW264.7 macrophages and SVEC4-10 endothelial cells. Macrophages or endothelial cells pre-incubated with HMB-TZD for 2 h and then stimulated with lipopolysaccharide or tumor necrosis factor-alpha (TNF-alpha) displayed reduced cytokine production. Also, HMB-TZD reduced cell migration and adhesion in accordance with decreased proinflammatory molecule production in vitro and ex vivo. HMB-TZD treatment of 8-week-old male Ldlr-/- mice resulted in significantly reduced atherosclerotic lesions without a change to plasma lipid profiles. Moreover, aortic expression of pro-atherogenic molecules involved in the recruitment of monocytes to the aortic wall, including TNF-alpha , MCP-1, and VCAM-1, was downregulated. HMB-TZD also reduced macrophage infiltration into atherosclerotic lesions. In conclusion, HMB-TZD ameliorates atherosclerotic lesion formation possibly by reducing the expression of proinflammatory molecules and monocyte/macrophage recruitment to the lesion. These results suggest that HMB-TZD, and benzylidenethiazole analogs in general, may have therapeutic potential as treatments for atherosclerosis.


Subject(s)
Animals , Atherosclerosis/drug therapy , Cell Adhesion/drug effects , Cell Line , Cell Movement/drug effects , Chemokine CCL2/metabolism , Dinoprostone/metabolism , Enzyme-Linked Immunosorbent Assay , Humans , Leukotriene B4/metabolism , Macrophages/cytology , Male , Mice , Monocytes/cytology , Random Allocation , Receptors, LDL/deficiency , Thiazolidinediones/therapeutic use , Tumor Necrosis Factor-alpha/pharmacology
9.
Article in English | WPRIM | ID: wpr-38821

ABSTRACT

BACKGROUND/AIMS: P2/MS is a noninvasive marker for detecting hepatic fibrosis in patients with viral hepatitis. However, the applicability of P2/MS in patients with nonalcoholic fatty liver disease (NAFLD) has not yet been validated. This study aimed to validate P2/MS and compare it to other noninvasive fibrosis scoring systems in Korean patients with NAFLD. METHODS: Consecutive patients who underwent liver biopsy between January 2002 and December 2009 at Seoul National University Hospital, Seoul, Korea were enrolled in this study. Fibrosis stage was determined using the METAVIR scoring system. RESULTS: A total of 235 patients were included in the study: advanced fibrosis (METAVIR F3-F4) was present in 7 patients. No patient was over-staged among 162 patients with a P2/MS score above the high cut-off (95), resulting in a high negative predictive value (NPV) of 100% (95% confidence interval, 97.1-100). There was no significant difference between the area under the receiver-operating characteristic curve (AUROC) of the FIB-4 (0.964) and the AUROC of the NAFLD fibrosis score (0.964) or P2/MS (0.940) for detecting advanced fibrosis. If P2/MS was implemented in the Korean patients with NAFLD, 68.9% of liver biopsies might be avoided. CONCLUSIONS: P2/MS has a high NPV for excluding advanced fibrosis in Korean patients with NAFLD, and can reduce the burden of liver biopsy in the majority of cases. Since there were few patients with advanced fibrosis, further studies are warranted in a cohort including more patients with advanced fibrosis to validate the low cut-off value.


Subject(s)
Adult , Aged , Alanine Transaminase/blood , Area Under Curve , Aspartate Aminotransferases/blood , Blood Cell Count , Diagnosis, Differential , Fatty Liver/complications , Female , Humans , Liver Cirrhosis/complications , Male , Middle Aged , Monocytes/cytology , Neutrophils/cytology , Platelet Count , Predictive Value of Tests , ROC Curve , Republic of Korea , Severity of Illness Index
10.
Yonsei Medical Journal ; : 990-998, 2011.
Article in English | WPRIM | ID: wpr-30290

ABSTRACT

PURPOSE: Dendritic cell (DC) vaccination for melanoma was introduced because melanoma carries distinct tumor-associated antigens. The purpose of this study was to investigate the efficacy and safety of DC vaccination for melanoma in Korea. MATERIALS AND METHODS: Five patients with stage IV and one with stage II were enrolled. Autologous monocyte-derived DCs (MoDCs) were cultured and pulsed with tumor-lysate, keyhole limpet hemocyanin, and cytokine cocktail for mature antigen-loaded DC. DC vaccination was repeated four times at 2-week intervals and 2-4x107 DC were injected each time. RESULTS: Reduced tumor volume was observed by PET-CT in three patients after DC vaccination. Delayed type hypersensitivity responses against tumor antigen were induced in five patients. Tumor antigen-specific IFN-gamma-producing peripheral blood mononuclear cells were detected with enzyme-linked immunosorbent spot in two patients. However, the overall clinical outcome showed disease progression in all patients. CONCLUSION: In this study, DC vaccination using tumor antigen-loaded, mature MoDCs led to tumor regression in individual melanoma patients. Further standardization of DC vaccination protocol is required to determine which parameters lead to better anti-tumor responses and clinical outcomes.


Subject(s)
Dendritic Cells/cytology , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunotherapy/methods , Melanoma/therapy , Monocytes/cytology , Treatment Outcome
11.
Article in English | WPRIM | ID: wpr-8329

ABSTRACT

BACKGROUND/AIMS: P2/MS is known as a simple, accurate, and noninvasive marker for determination of the degree of hepatic fibrosis in patients with viral hepatitis. We aimed to validate P2/MS in patients with HCC. METHODS: Consecutive HCC patients who underwent surgical resection between June 2007 and March 2009 at Seoul National University Hospital were enrolled. Fibrosis stage was reviewed and assessed according to METAVIR scoring. P2/MS values [platelet count (109/L)]2/[monocyte fraction (%)xsegmented neutrophil fraction (%)] and other noninvasive fibrosis scoring systems were calculated. RESULTS: A total of 171 patients were included; seven patients with METAVIR F1, 31 with F2, 41 with F3, and 92 with F4. The area under the receiver-operating characteristic curve of P2/MS was 0.804 [95% confidence interval (CI), 0.681~0.927] for detection of significant fibrosis (F2-F4) and 0.769 (95% CI, 0.698~0.839) for detection of histological cirrhosis (F4). At a value 115, P2/MS ruled out significant fibrosis with a sensitivity of 90.2% (95% CI, 84.4~94.1) and a negative likelihood ratio of 0.34 (95% CI, 0.106~0.095). P2/MS had a superior efficacy for detection of hepatic fibrosis in patients with HCC compared to the other noninvasive panels. CONCLUSIONS: P2/MS can accurately detect fibrosis in patients with HCC. Thus, P2/MS might be utilized as a noninvasive index reflecting the degree of hepatic fibrosis in HCC patients.


Subject(s)
Aged , Area Under Curve , Carcinoma, Hepatocellular/complications , Cohort Studies , Female , Health Status Indicators , Humans , Liver Cirrhosis/complications , Liver Neoplasms/complications , Male , Middle Aged , Monocytes/cytology , Neoplasm Staging , Neutrophils/cytology , Platelet Count , ROC Curve , Reproducibility of Results , Retrospective Studies , Severity of Illness Index
12.
Article in English | WPRIM | ID: wpr-196694

ABSTRACT

Small heterodimer partner (SHP) is an atypical member of nuclear receptor superfamily that lacks a DNA-binding domain. In previous study, we showed that SHP, c-jun, p65 of NF-kappaB subunits, and p21WAF1 expression was increased during monocytic differentiaton with the exposure of human leukemia cells to a differentiation agent, PMA. In this study, c-Jun and p65 were shown to mediate the transcriptional activation of the SHP promoter. In addition, SHP induced the cell cycle regulatory protein levels and cooperatively increased an induction of p21WAF1 expression with p65. Furthermore, SHP protected differentiated cells from etoposide-induced cellular apoptosis through the induction and cytoplasmic sequestration of p21WAF1. Complex formation between SHP and p21WAF1 was demonstrated by means of coimmunoprecipitation. These results suggest that SHP prolongs a cellular survival of differentiating monocytes through the transcriptional regulation of target genes of cell survival and differentiation.


Subject(s)
Apoptosis , Cell Differentiation , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p21/genetics , Gene Expression Regulation , Humans , Monocytes/cytology , Promoter Regions, Genetic , Proto-Oncogene Proteins c-jun/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Transcription Factor RelA/genetics
13.
Article in English | WPRIM | ID: wpr-15564

ABSTRACT

Flow cytometry was used to identify and characterize monoclonal antibodies (mAbs) that react with rabbit leukocyte differentiation molecules (LDM). Screening sets of mAbs, developed against LDM in other species, for reactivity with rabbit LDM yielded 11 mAbs that recognize conserved epitopes on rabbit LDM orthologues and multiple mAbs that recognize epitopes expressed on the major histocompatibility class I or class II molecules. Screening of mAbs submitted to the Animal Homologues Section of the Eighth Human Leukocyte Differentiation Workshop yielded 7 additional mAbs. Screening of mAbs generated from mice immunized with leukocytes from rabbit thymus or spleen or concanavalin A activated peripheral blood and/or spleen lymphocytes has yielded 42 mAbs that recognize species restricted epitopes expressed on one or more lineages of leukocytes. Screening of the anti-rabbit mAbs against leukocytes from other species yielded one additional mAb. The studies show that screening of existing sets of mAbs for reactivity with rabbit LDM will not be productive and that a direct approach will be needed to develop mAbs for research in rabbits. The flow cytometric approach we developed to screen for mAbs of interest offers a way for individual laboratories to identify and characterize mAbs to LDM in rabbits and other species. A web-based program we developed provides a source of information that will facilitate analysis. It contains a searchable data base on known CD molecules and a data base on mAbs, known to react with LDM in one or more species of artiodactyla, equidae, carnivora, and or lagomorpha.


Subject(s)
Animals , Antibodies, Monoclonal/immunology , Antigens, Differentiation/metabolism , B-Lymphocytes/cytology , Basophils/cytology , Epitopes/genetics , Flow Cytometry , Gene Expression Regulation , Granulocytes/cytology , Leukocytes/immunology , Mice , Monocytes/cytology , Rabbits , T-Lymphocytes/cytology
14.
Saudi Medical Journal. 2007; 28 (5): 688-695
in English | IMEMR | ID: emr-85099

ABSTRACT

To find out the effects of hepatocyte growth factor [HGF] in the development of dendritic cells [DC] from the peripheral monocytes. The study was carried out in Black Sea Technical University Hospital, Trabzon, Turkey between 2003-2004. Seven different cytokine combinations were employed to assess phenotypical and functional differences of DCs from the peripheral monocytes in serum free culture media. Peripheral monocytes were incubated in media with cytokines for 5 days. The tumor necrosis factor-alpha [TNF-alpha] was added to the cell culture on day 5 and incubated for another 2 days. Surface and co-stimulating molecules on the cells were assessed by flowcytometry. The functional capacity of the DCs was evaluated on day 7 by purified protein derivative loading and subsequent lymphoproliferation test using methyl tetrazolium staining. On the 5th day of incubation DC development was observed in all cytokine groups, but cells were superior in cultures maintained in the presence of interleukin-4 combinations with granulocyte-macrophage colony stimulating factor [GM-CSF] or with GM-CSF+HGF. Moreover, the expression of surface and co-stimulating molecules increased significantly after incubation with TNF-alpha. The effect of PPD loaded-DCs on proliferation of lymphocytes was more striking in HGF containing groups. It was concluded that HGF supplemented cultures exert some additive effects in relation to function of monocyte-derived DCs. But HGF alone does not seem to augment monocyte-derived DC proliferation and maturation significantly


Subject(s)
Humans , Cell Differentiation/physiology , Dendritic Cells/cytology , Monocytes/cytology , Cells, Cultured
15.
Article in English | WPRIM | ID: wpr-37558

ABSTRACT

One of characteristic features of AIDS-related encephalitis and dementia is the infiltration of monocytes into the CNS. HIV-1 Tat was demonstrated to facilitate monocyte entry into the CNS. In this study, we examined the effect of HIV-1 Tat on the expression of adhesion molecules, generation of reactive oxygen species (ROS) and NF-kappaB activation in CRT-MG human astroglioma cells. Treatment of CRT-MG cells with HIV-1 Tat protein significantly increased protein and mRNA levels of ICAM-1 and VCAM-1, as measured by Western blot analysis and RT-PCR, indicating that Tat increases these protein levels at an mRNA level. In addition, Tat induced the activation of NF-kappaB in astrocytes. Treatment of CRT-MG with NF-kappaB inhibitors led to decrease in Tat-induced protein and mRNA expression of ICAM-1 and VCAM-1. Furthermore, HIV-1 Tat protein increased ROS generation. Inhibition of Tat-induced ROS generation by N-acetyl cysteine, vitamin C and diphenyl iodonium suppressed Tat-induced NF-kappaB activation, ICAM-1 and VCAM-1 expression, and monocyte adhesion in CRT-MG. These data indicate that HIV-1 Tat can modulate monocyte adhesiveness by increasing expression of adhesion molecules such as ICAM-1 and VCAM-1 via ROS- and NF-kappaB-dependent mechanisms in astrocytes.


Subject(s)
Vascular Cell Adhesion Molecule-1/genetics , Up-Regulation/drug effects , Transcription, Genetic/genetics , Reactive Oxygen Species/metabolism , NF-kappa B/metabolism , Monocytes/cytology , Intercellular Adhesion Molecule-1/genetics , Humans , HIV-1 , Gene Products, tat/pharmacology , Cell Line , Cell Adhesion/drug effects , Astrocytes/cytology
16.
Article in English | WPRIM | ID: wpr-21101

ABSTRACT

HIV-1 Tat is considered to be one of key players to facilitate monocyte entry into the CNS, which is characteristic feature of AIDS-related encephalitis and dementia. This study was performed to determine the regulatory function of superoxide dismutase (SOD) on the HIV-1 Tat-induced signaling pathways leading to NF-kappaB activation, expression of adhesion molecules, and monocyte adhesion in CRT-MG human astroglioma cells by using cell-permeable SOD. When cell-permeable SOD was added to the culture medium of CRT-MG cells, it rapidly entered the cells in dose- and time-dependent manners. Treatment of astrocytes with cell-permeable SOD led to decrease in Tat-induced ROS generation as well as NF-kappaB activation. Cell-permeable SOD inhibited the activation of MAP kinases including ERK, JNK and p38 by HIV-1 Tat. Treatment of CRT-MG cells with cell-permeable SOD significantly inhibited protein and mRNA levels of ICAM-1 and VCAM-1 up-regulated by HIV-1 Tat, as measured by Western blot analysis and RT-PCR. Furthermore, enhanced adhesiveness of monocyte to astrocyte by HIV-1 Tat was significantly abrogated by pretreatment with cell-permeable SOD fusion proteins. These data indicate that SOD has a regulatory function for HIV-1 Tat-induced NF-kappaB activation in astrocytes and suggest that cell-permeable SOD can be used as a feasible therapeutic agent for regulation of ROS-related neurological diseases.


Subject(s)
Astrocytes/enzymology , Cell Adhesion/physiology , Cell Membrane Permeability , Gene Products, tat/pharmacology , HIV Infections/metabolism , HIV-1/chemistry , Humans , Monocytes/cytology , Signal Transduction , Superoxide Dismutase/genetics
17.
Article in English | IMSEAR | ID: sea-23340

ABSTRACT

BACKGROUND & OBJECTIVE: The understanding of the pathogenesis of psoriasis vulgaris has focused on T cell mediated immune disorder for many years. Recent studies provide evidence that dendritic cells may be of major importance as regulatory cells driving the psoriasis tissue reaction, and they are one of the therapeutic targets. In order to further characterize the role of dendritic cells in psoriasis, this study was designed to assess the differentiation of dendritic cells from monocytes (MoDC), the expression of phagocytosis related receptors by MoDC, their endocytic activity for fluorescent beads and lucifer yellow as well as their superoxide generation in patients with psoriasis. METHODS: Twenty eight patients with psoriasis vulgaris and 12 healthy controls were included in the study. MoDC were obtained by culturing monocytes with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 5 days. Cell surface expression of CD1a, CD14, CD40, CD80, CD83, CD86, HLA-DR, mannose receptor (MR) and Fcg receptors by MoDC and their endocytosis of dextran and lucifer yellow were analyzed by flow cytometry. Zymosan ingestion was measured to access the phagocytosis of MoDC. RESULTS: Differentiation of monocytes to dendritic cells was upregulated in patients manifested as significantly increased expression of CD40, CD80, CD86 and HLA-DR compared with that in healthy controls (P<0.01). Expression of MR and Fcg receptor II (CD32) by MoDC was significantly increased in patients with psoriasis as well (P<0.01). Endocytosis of dextran but not lucifer yellow in patients was significantly higher than controls (P<0.01), and significantly enhanced phagocytosis by increasing zymosan ingestion was also observed (P<0.01) in patients. Taken together, endocytic and phagocytic activity of MoDC in psoriasis was increased than normal persons. INTERPRETATION & CONCLUSION: Enhanced activity of dendritic cells binding and capturing foreign antigens for subsequent antigen presentation and the initiation of immune responses in psoriasis may contribute to the pathogenesis of the disease. The upregulated expression of MR and the enhanced endocytic activity of DC might be an explanation for the absence of skin infection observed in psoriasis.


Subject(s)
Adolescent , Adult , Aged , Biomarkers/metabolism , Cell Differentiation/physiology , China , Dendritic Cells/cytology , Endocytosis/physiology , Female , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Humans , Lectins, C-Type/immunology , Male , Mannose-Binding Lectins/immunology , Middle Aged , Monocytes/cytology , Phagocytosis/physiology , Psoriasis/immunology , Random Allocation , Receptors, Cell Surface/immunology , Receptors, IgG/immunology
18.
Article in English | WPRIM | ID: wpr-18469

ABSTRACT

Human 8-oxo-G-DNA glycosylase 1 (hOGG1) is a DNA glycosylase to cleave 8-oxo-7,8-dihydroguanine (8-oxo-G), a mutagenic DNA adduct formed by oxidant stresses. Here, we examined hOGG1 protein expression and repair activity to nick a DNA strand at the site of 8-oxo-G during differentiation of hematopoietic cells using HL-60 cells. Overall expression of hOGG1 protein was increased during granulocytic differentiation of HL-60 cells induced by DMSO and monocytic differentiation by vitamine D3. Greater level of hOGG1 protein was expressed in DMSO-treated cells. However, change in the DNA nicking activity was not in parallel with the change in hOGG1 protein expression, especially in PMA-treated cells. In PMA- treated cells, the level of hOGG1 protein was lowered, even though the DNA nicking activity was elevated, in a manner similar to the changes in serum- deprived HL-60 cells. These results indicate that hOGG1 expression change during differentiation of hematopoietic stem cells for adaptation to new environments. And the DNA cleaving activity may require additional factor(s) other than expressed hOGG1 protein, especially in apoptotic cell death.


Subject(s)
Blotting, Western , Cell Differentiation , Culture Media, Serum-Free/pharmacology , DNA Glycosylases/metabolism , Enzyme Activation , Gene Expression Regulation, Enzymologic/drug effects , Granulocytes/cytology , HL-60 Cells , Humans , Monocytes/cytology
19.
Asian Pac J Allergy Immunol ; 2001 Jun; 19(2): 85-91
Article in English | IMSEAR | ID: sea-36695

ABSTRACT

Kawasaki disease (KD) is a childhood-onset vascular disease. We assessed the concentrations of macrophage-colony stimulating factor (M-CSF) and those of lipids in sera from patients with KD. The M-CSF concentration in patients with acute-phase KD was 2,914+/-159 U/ml, significantly higher than that in control subjects with Infectious diseases (1,241+/-96 U/ml). The elevated levels of this cytokine in the acute phase fell to 1,319+/-138 U/ml in the convalescent phase. Total and high-density lipoprotein cholesterol concentrations in acute phase KD (113.8+/-8.4 and 21.5+/-2.3 mg/dl, respectively) were lower than in the infectious disease controls (195.8+/-7.0 and 62.5+/-1.8 mg/dl). The elevation of M-CSF correlated with the decrease of total and high-density lipoprotein cholesterol. Overproduction of macrophage-colony stimulating factor activates macrophages and monocytes and may disturb the lipid metabolism. Both effects could contribute to vasculitis in KD.


Subject(s)
Child Welfare , Child, Preschool , Cholesterol/blood , Cholesterol, HDL/blood , Female , Humans , Immunoglobulins, Intravenous/therapeutic use , Infant , Infant Welfare , Japan/epidemiology , Leukocyte Count , Macrophage Colony-Stimulating Factor/blood , Male , Monocytes/cytology , Mucocutaneous Lymph Node Syndrome/blood , Statistics as Topic
20.
Medicina (B.Aires) ; 60(2): 202-10, 2000. tab, graf
Article in English | LILACS | ID: lil-262212

ABSTRACT

Resting human T cells are known to express significant numbers of intermediate but none or barely detectable low and high a affinity IL-2 receptors (IL-2R). IL-2 alone failed to induce proliferation in these cells, However, in presence of small proportion of autologous monocytes, as low as 22 pM, IL-2 induced high levels of proliferation in resting T cells. Introduction of a semi permeable between the two cell types or addition of an anti-CD 11b mAb inhibited such induction of proliferation by IL-2. Neither recombinant IL-1 por IL-1 containing cell-free extracts from activated monocytes substituted for intact monocytes. Autologous B cells failed to replace monocytes. Using antigen-specific cloned human T cells we have shown a lack of requirement for antigen. The proliferation was inhibited by anti-IL-2R alpha mAb. IL-2 appears to be unique since neither IL-4 nor IL-6, alone or in presence of monocytes, led to induction of proliferation in resting T cells. Combination of IL-2 and monocytes induced proliferation in all T cell subpopulations (CD4, CD8, CD45RA and CD45RO) and antigen-specific clones examined. It also induces mRNA and surface expression of IL-2R alpha, appearance of high affinity IL-2R and induction of proliferation in large proportions of T cells. As in humans, the IL-2 induction of proliferation in murine resting T cell required contact with syngeneic monocytes, suggesting that such a mechanism of cells activation is highly conserved.


Subject(s)
Humans , Animals , Mice , Interleukin-2/pharmacology , Lymphocyte Activation/drug effects , Monocytes/physiology , Receptors, Interleukin-2/physiology , T-Lymphocytes/drug effects , Cell Culture Techniques , Interferon-alpha/pharmacology , Interleukin-2/physiology , Mice, Inbred BALB C , Monocytes/cytology , Thymidine
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