Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have investigated constitutive and phytohaemagglutinin (PHA) + phorbol 12-myristate 13-acetate (PMA)-induced gene expression of tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-2, IL-3, IL-4, IL-10, IL-12 and granulocyte macrophage colony-stimulating factor (GM-CSF) in peripheral blood mononuclear cells (PBMCs) of 10 patients with Takayasu's arteritis (TA) and 10 healthy controls by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The constitutive mRNA expression of TNF-alpha (69.0 +/- 4.0%versus 27.5 +/- 18.0%; P = 0.001) and IL-4 (60.0 +/- 10.0%versus 0%; P = 0.001) was significantly higher in patients than controls; that of IL-3 was comparable in both groups (38.0 +/- 6.0%versus 32.0 +/- 5.0%; P = 0.651) while no constitutive mRNA expression was observed for the other cytokines studied. The stimulated PBMCs of patients, as compared with the controls, had higher mRNA gene expression of TNF-alpha (127.0 +/- 16.0%versus 54.0 +/- 6.0%; P = 0.001), IFN-gamma (93.0 +/- 13.0%versus 57.0 +/- 5.0%; P = 0.032), IL-2 (109.0 +/- 13.0%versus 68.0 +/- 6.0%; P = 0.015), IL-3 (60.0 +/- 8.0%versus 21.2 +/- 3.0%; P = 0.045) and IL-4 (68.0 +/- 7.0%versus 27.0 +/- 7.2%; P = 0.01) The mRNA expression of IL-10 was lower in patients than controls (35.0 +/- 8.0%versus 75.0 +/- 12.0%; P = 0.022). The GM-CSF mRNA was similar (102.0 +/- 6.0%versus 89.0 +/- 5.0%; P = 0.475) in both groups. Stimulation of cells with PHA + PMA showed no IL-12 expression but stimulation with lipopolysaccharide induced higher IL-12 mRNA in patients than controls (83.0 +/- 14.0%versus 33.0 +/- 4.0%; P = 0.005). Our data suggest that an inflammatory cytokine signature exists in TA with a key role for TNF-alpha, IL-4, IL-10 and IL-12 in different pathological processes of the disease.
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PMID:Cytokine mRNA repertoire of peripheral blood mononuclear cells in Takayasu's arteritis. 1549 51

We recently identified a reduction in the neutrophil surface expression of common beta chain (beta c) of the receptor for granulocyte macrophage-colony stimulating factor (GM-CSF) in the patients with myelodysplastic syndromes (MDS). To determine the etiology of the impaired beta c expression, beta c mRNA from neutrophilic granulocytes of MDS patients and healthy controls was analyzed by a combination of direct reverse transcriptase-polymerase chain reaction-based single-strand conformational polymorphism and sequencing. Nine different beta c transcripts were detected, but none was specific for MDS. However, one of the transcripts (beta c79) containing a 79-base intron insertion between exons V and VI was significantly increased in MDS. This 27-kd isoform consisted of the beta c N-terminal 182 amino acids followed by a new 84-amino-acid sequence. beta c79 was overexpressed in all MDS subtypes. No genomic mutations were detected within the intron or at the intron/exon boundaries. The isoform is predominantly located in the cytoplasm by Western blot analysis and was unable to generate high-affinity binding sites or transduce a signal for proliferation when coexpressed with the receptor for human GM-CSF alpha chain. Our study suggests that the accumulation of the abnormal beta c transcripts with intron V retention results in the reduction in cell-surface expression of beta c observed in MDS.
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PMID:Accumulation of an intron-retained mRNA for granulocyte macrophage-colony stimulating factor receptor common beta chain in neutrophils of myelodysplastic syndromes. 1572 48

Diesel exhaust particles (DEPs) at three concentrations (5, 35, and 50 mg/kg body weight) were instilled into rats intratracheally. We studied gene expression at 1, 7, and 30 days postexposure in cells obtained by bronchoalveolar lavage (BAL) and in lung tissue. Using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), we measured the mRNA levels of eight genes [interleukin (IL)-1beta, IL-6, IL-10, iNOS (inducible nitric oxide synthase), MCP-1 (monocyte chemoattractant protein-1), MIP-2 (macrophage inflammatory protein-2), TGF-beta1 (transforming growth factor-beta1), and TNF-alpha (tumor necrosis factor-alpha )] in BAL cells and four genes [IL-6, ICAM-1 (intercellular adhesion molecule-1), GM-CSF (granulocyte/macrophage-colony stimulating factor), and RANTES (regulated upon activation normal T cell expressed and secreted)] in lung tissue. In BAL cells on day 1, high-dose exposure induced a significant up-regulation of IL-1beta, iNOS, MCP-1, and MIP-2 but no change in IL-6, IL-10, TGF-beta1, and TNF-alpha mRNA levels. There was no change in the mRNA levels of IL-6, RANTES, ICAM-1, and GM-CSF in lung tissue. Nitric oxide production and levels of MCP-1 and MIP-2 were increased in the 24-hr culture media of alveolar macrophages (AMs) obtained on day 1. IL-6, MCP-1, and MIP-2 levels were also elevated in the BAL fluid. BAL fluid also showed increases in albumin and lactate dehydrogenase. The cellular content in BAL fluid increased at all doses and at all time periods, mainly due to an increase in polymorphonuclear leukocytes. In vitro studies in AMs and cultured lung fibroblasts showed that lung fibroblasts are a significant source of IL-6 and MCP-1 in the lung.
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PMID:Time course of gene expression of inflammatory mediators in rat lung after diesel exhaust particle exposure. 1586 72

Polymorphonuclear leukocytes have been shown to use a multitude of effector functions to combat pathogens and tumors, including enzymes, defensins, and toxic products such as oxygen radicals and nitrogen oxides. Recent studies provided evidence for the expression of granzymes (gzms) and perforin (perf) within the cytotoxic arsenal of human neutrophils, the validity of which was questioned by 2 subsequent studies. We have now used cytology, intracellular flow cytometry, enzymatic assays, immunoelectron microscopy, and quantitative reverse transcriptase-polymerase chain reaction to obtain evidence of the presence of gzms and/or perf in mouse Gr-1+ granulocyte populations. The data obtained clearly demonstrate that neither in vitro- nor in vivo-derived mouse granulocytes synthesize gzmA and gzmB or perf, even following infection/immunization with pathogens or pathogen-derived material. A parallel comparable analysis on the expression of gzmB in human neutrophils from 3 healthy control subjects and 4 patients with diverse diseases failed to detect gzmB expression. The data indicate that polymorphonuclear leukocytes from mice and humans lack the 3 cytotoxic effector molecules, gzmA, gzmB, and perf, generally associated with natural killer and cytotoxic T lymphocytes.
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PMID:Quiescent and activated mouse granulocytes do not express granzyme A and B or perforin: similarities or differences with human polymorphonuclear leukocytes? 1599 31

We investigated the effects of immunizing with several genes and subtypes of HIV-1. The genes used as immunogens were: gp160 envelope (env subtypes A, B and C), p37gag (gag subtypes A and B), rev (subtype B) and reverse transcriptase (RT subtype B). The different genes are all carried by separate plasmids. C57BL/6 and BALB/c mice were immunized with different combinations of the genes together with recombinant cytokine granulocyte macrophage-colony stimulating factor. The env genes injected alone induced significantly stronger cellular responses to envelope in both strains of mice than when env genes were injected together with gag and RT genes. In the C57BL/6 mice, the envelope specific responses were significantly increased after spatial separation of env genes from gag and RT genes as compared to when all vaccine genes were injected as a mixture. The gag responses were strong in gag-immunized animals and were not significantly affected by the spatial separation of gag and RT genes from the env genes. Our results illustrate the importance of being cautious when formulating multivalent genetic vaccines and that it might be possible to overcome lost immune responses through spatial separation of vaccine antigens.
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PMID:Reduced cellular immune responses following immunization with a multi-gene HIV-1 vaccine. 1617 43

The purpose of this study was to investigate the influence of the cytostatic drug, 5-fluorouracil (5-FU), which causes depletion of heterophil granulocytes, on clinical symptoms and histological lesions during the progress of infectious bursal disease virus (IBDV) infection in chickens. The aim was to disclose the mechanism behind the clinical disease symptoms. Three groups of specific pathogen free chickens were used for the experiment. Chickens in groups 1 and 3 were pretreated with 5-FU, while chickens in group 2 were treated with a placebo. After 5 days, the chickens in groups 2 and 3 were inoculated with the classical IBDV strain F52/70. Bursae of Fabricius were sampled at fixed intervals, and the progress of the infection was monitored by various histological techniques and reverse transcriptase-polymerase chain reaction (RT-PCR). We found correlation between histological observations and RT-PCR results. In the 5-FU pretreated chickens, IBDV caused only mild clinical symptoms, even though histological alterations similar to alterations caused by IBDV were still observed. The 5-FU pretreatment resulted in severe heterophil granulocyte depletion by days 2 and 3 after infection (post inoculation) and increased numbers of bursal secretory dendritic cells in the medulla of the follicles. IBDV infection seemed to induce fusion of secretory dendritic cells, resulting in formation of multinucleated giant cells, loaded with apoptotic B cells and virus particles associated with granules of bursal secretory dendritic cells. Our results indicate that the heterophil granulocytes together with the bursal secretory dendritic cells contribute to the outbreak and/or progress of clinical symptoms.
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PMID:Impact of heterophil granulocyte depletion caused by 5-fluorouracil on infectious bursal disease virus infection in specific pathogen free chickens. 1685 50

A variety of extraimmune system factors, including hormones, play a critical role in regulating immunity. Progesterone has been shown to affect immunity in rodents and humans, mainly at concentrations commensurate with pregnancy. These effects are primarily mediated via the progesterone receptor (PR), which acts as a transcription factor, although non-genomic effects of PR activation have been reported. In this study, we evaluated the effects of progesterone on rat dendritic cells (DCs) at ranges encompassing physiologic and pharmacologic concentrations to determine whether progesterone plays a role in modulating DC-mediated immune responses. DCs were derived by culturing rat bone marrow cells in granulocyte macrophage colony-stimulating factor and IL-4. Cells were analyzed for expression of PR using FACS analysis, real-time reverse transcriptase-PCR and fluorescent microscopy. Progesterone treatment of LPS-activated, mature bone marrow-derived dendritic cells (BMDCs) suppressed production of the pro-inflammatory response-promoting cytokines tumor necrosis factor-alpha and IL-1beta in a dose-dependent manner but did not affect production of the pro-inflammatory response-inhibiting cytokine IL-10. Treatment of cells with progesterone also resulted in down-regulation of co-stimulatory molecule CD80 and MHC class II molecule RT1B expression. In addition, progesterone inhibited DC-stimulated proliferation of T cells. Suppression of pro-inflammatory response-promoting cytokine production by progesterone was prevented using the PR antagonist RU486. There was no dose-dependent effect of progesterone treatment on immature DC capacity to take up antigenic peptide. These data indicate that progesterone directly inhibits mature rat BMDC capacity to drive pro-inflammatory responses. This mechanism could contribute to or account for some of the differential expression of autoimmune/inflammatory disease in females.
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PMID:Progesterone inhibits mature rat dendritic cells in a receptor-mediated fashion. 1728 56

PU.1, which is a transcription factor, promotes the terminal differentiation of alveolar macrophages (AMs). Its expression is regulated by granulocyte/macrophage colony-stimulating factor (GM-CSF). In this study of AMs in newborn rats, we performed immunohistochemical staining, acid phosphatase staining, reverse transcriptase polymerase chain reaction (RT-PCR), quantitative real-time PCR, cytokine assay, and electron microscopy. AMs at 3 and 7 days after birth had a large foamy appearance with an intracytoplasmic accumulation of surfactants. Weak expression of PU.1 was observed in the nuclei. AMs at 15 days after birth were smaller, and PU.1 expression had increased. Ultrastructurally, AMs at 1 day after birth had a smooth surface and abundant lamellar structures in the cytoplasm, whereas AMs at 56 days after birth were characterized by (1) abundant microvillar projections on the cell surface, and (2) well-developed lysosomes and a few lamellar structures in the cytoplasm. Acid phosphatase activity and the expression of mannose receptor, scavenger receptor, and GM-CSF receptor alpha were enhanced in AMs with time after birth. These results suggest that AMs are initially immature, and that their terminal differentiation starts after birth concomitantly with an increased expression of PU.1.
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PMID:Expression of PU.1 and terminal differentiation of alveolar macrophages in newborn rats. 1740

We have shown for the first time that pluripotent/multipotent stem cells can be isolated from blood in a relatively simple cell separation, which is fast and compatible with current standards. We used the germ line- and pluripotent-specific DAZL gene as a marker to demonstrate its use for identifying and isolating pluripotent/multipotent cells from blood. DAZL-expressing (DE) cells were identified in about 0.3% of umbilical cord blood (UCB) mononuclear cells. These DE cells do not express either blood cell differentiation markers, such as CD38, CD3, and CD14, or the blood progenitor cell markers, such as CD34 and CD133. Nevertheless, they express pluripotent embryonic stem (ES) cells OCT-4 and SOX-2 genes. To examine whether the DE cells exhibit stem cell characteristics, we seeded 10(3) DE cells in methylcellulose containing the ingredients needed for hematopoietic cell growth. Although DE cells are CD34-, a mean (+/-SD) of 21 +/- 4 colonies were formed per plate, of which 4.8% were colony-forming unit granulocyte, erythroid, macrophage, megakaryocytes (CFU-GEMMs). Mononuclear or CD34+ cells isolated from UCB formed 6 +/- 1 and 53 +/- 8 colonies per plate, respectively, of which 0% and 3.8% were CFU-GEMMs. DE cells grown in methylcellulose without cytokines formed various colonies, which demonstrated expression pattern that is typical of neurons, endothelial, bone/cartilage, cardiomyocyte, and hepatocyte differentiation as determined by reverse transcriptase-PCR (RT-PCR). Our results suggest that DE cells, which possess some pluripotent/multipotent characteristics of ES cells, can be easily isolated from blood. These cells may be effective in various therapeutic applications with no biological or ethical ramifications.
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PMID:A method for isolating pluripotent/multipotent stem cells from blood by using the pluripotent and germ-line DAZL gene as a marker. 1932 14

Epidermal mRNA for interleukin 1beta (IL-1beta) has been shown to be increased following exposure of mouse skin to sensitizing compounds. In addition, this early upregulation of IL-1beta was specific for contact sensitizers, while expression of IL-1beta was unaffected by irritants. Langerhans cells are the major source of IL-1beta within the epidermis in the induction phase of skin sensitization. Since the isolation of Langerhans cells from skin biopsies results only in low frequencies, we decided to use dendritic cells (DCs) generated from peripheral blood as Langerhans cell equivalents to investigate the ability of five contact sensitizers and one irritant to induce IL-1beta gene expression in vitro. For our studies we cultivated DCs in serum-free medium supplemented with granulocyte/macrophage-colony stimulation factor (GM-CSF) and interleukin 4 (IL-4). The DCs showed a typical dendritic morphology, a characteristic expression of surface markers and high stimulatory capacity for autologous T cells. 5-day-old DCs were incubated with subtoxic concentrations of the contact sensitizers pentadecyl-catechol, 2,4,6-trinitrobenezene sulfonic acid, 2,4-dinitrofluorobenzene, NiSO(4), K(2)Cr(2)O(7) and the irritant sodium dodecyl sulfate. IL-1beta mRNA expression was detected by using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique and non-radioactive hybridization procedures. For all contact sensitizers, expression of IL-1beta mRNA increased, whereas treatment with the irritant SDS had no significant effect on IL-1beta expression. Thus we developed an in vitro system, which may be useful to evaluate allergic potentials of chemicals and products.
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PMID:In vitro model for contact sensitization: II. Induction of IL-1beta mRNA in human blood-derived dendritic cells by contact sensitizers. 2065 60


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