[Cytokine measurements from patient specimens].
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Cytokines↗
Biomedical subjects
M Hurme
Publications and source records attributed to M Hurme.
Induction of transforming growth factor beta 1 and its receptor expression during myeloid leukemia cell differentiation.
The human myeloid leukemia cell lines HL-60, U-937, and THP-1 were used to analyze the alterations of transforming growth factor beta (TGF-beta) during hematopoietic cell growth and differentiation. Differentiation of these cell lines was induced by the phorbol ester phorbol 12-myristate 13-acetate, tumor necrosis factor alpha or by retinoic acid. Northern hybridization analyses indicated that the basal levels of TGF-beta 1, latent TGF-beta binding protein, and type II TGF-beta receptor (T beta IIR) mRNAs were low in untreated cells. Major increases of these mRNAs were observed in cells treated with phorbol 12-myristate 13-acetate, with maximal induction after 12-72 h of stimulation. Retinoic acid and tumor necrosis factor alpha elevated significantly only the expression of T beta IIR mRNA. TGF-beta 1 induced its receptor mRNA in HL-60 and U937-1SF cells but not in THP-1 cells. These changes in gene expression were related to the differentiation of myeloid leukemia cells. Affinity labeling with 125I-TGF-beta 1 indicated that type I TGF-beta receptor was coregulated with T beta IIR. Types I and II receptors were coprecipitated by T beta IIR-specific antibodies. Differentiation of myeloid cells induced secretion of latent TGF-beta 1 protein, as shown by immunoblotting, but significant changes in the levels of active TGF-beta were not observed. These results indicate that the genes involved in TGF-beta signal transduction are coordinately up-regulated during myeloid differentiation.
Amino Acid Sequence↗
Activation of interleukin-1 beta gene expression during retinoic acid-induced granulocytic differentiation of promyeloid leukemia cells.
We have examined the expression of the interleukin 1 beta (IL-1 beta) gene during the granulocytic differentiation of two promyeloid leukemia cell lines, HL-60 and NB4. HL-60 is known to differentiate along the granulocytic pathway after treatment with 13-trans-retinoic acid (13-trans-RA), whereas treatment with phorbol myristate acetate (PMA) leads to development of mature macrophages. NB4 cells are derived from the bone marrow of an acute promyelocytic leukemia (APL) patient in relapse, have a translocated RA receptor-alpha, and are converted into nondividing granulocytes by 13-trans-RA treatment. When HL-60 or NB4 were cultured in the presence of 13-trans-RA, IL-1 beta mRNA and protein levels were increased. In the more mature THP-1 cells which are induced to macrophage-like cells by 13-trans-RA treatment, RA was unable to induce any IL-1 beta expression, implying that the effect of 13-trans-RA is associated with granulocytic differentiation. Moreover, PMA and 13-trans-RA had a strong synergistic effect in the induction of IL-1 beta gene expression. Nuclear run-off analysis indicated that the increased IL-1 beta gene expression was due to an enhanced rate of transcription. When the cells were transfected with an IL-1 beta-X-CAT reporter plasmid containing the -2982/-2748 promoter segment of the IL-1 beta gene conferring responsiveness to PMA, both NB4 and HL-60 cells responded with increased CAT activity when stimulated with 13-trans-RA alone. In contrast to PMA, 13-trans-RA was unable to increase AP-1 enhancer activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Base Sequence↗
Activation of NF-kappa B by cAMP in human myeloid cells.
Nuclear factor kappa B (NF-kappa B) is a DNA-binding regulatory factor that controls the transcription of a number of genes. Various agents are known to activate this factor. We have studied the ability of cAMP to stimulate NF-kappa B activity in human myeloid cells. Electrophoretic mobility assay revealed that structural cAMP analogs and agents elevating intracellular cAMP levels induced NF-kappa B DNA-binding activity. The inducibility was dependent on the maturation stage of myeloid cells. In promyelocytic HL-60 cells cAMP induced higher NF-kappa B activity than in more differential THP-1 cells and in human monocytes. By transfecting HL-60 and THP-1 cells with reporter constructs containing NF-kappa B DNA-binding sites, we observed that cAMP-induced NF-kappa B was transcriptionally active.
Base Sequence↗
Synergism between protein-kinase C and cAMP-dependent pathways in the expression of the interleukin-1 beta gene is mediated via the activator-protein-1 (AP-1) enhancer activity.
In many different cell types treatment with phorbol esters (e.g. 4 beta-phorbol 12-myristate 13-acetate, PMA) leads to the activation of protein-kinase C (PKC) and subsequently to the activation of the activator-protein-1(AP-1)-responsive gene expression. We have previously reported that a structural analog of cAMP (dibutyryl cAMP, Bt2cAMP) or agents elevating the endogenous cAMP levels strongly enhanced the PMA-induced interleukin-1 beta(IL-1 beta)-gene expression in human myeloid leukemia cells (THP-1, HL-60). We have now examined the role of AP-1 in the regulation of the IL-1 beta gene expression by PKC and cAMP in THP-1 cells. AP-1 is a complex composed of products of the jun and fos gene families. Our studies show that Bt2cAMP enhances the PMA-induced c-fos and jun-B expression, but inhibits c-jun expression. Electrophoretic mobility-shift assay revealed that Bt2cAMP also increased the PMA-induced AP-1 DNA-binding activity. The functional role of the increased AP-1 DNA-binding activity was studied by transfecting THP-1 cells with reporter constructs containing AP-1 sites [Col-TREx5/TK-CAT and IL-1 beta-X-CAT, which contains the putative 12-O-tetradecanoyl-phorbol-13-acetate(TPA)-responsive element of the IL-1 beta gene]. Transient transfection assay demonstrated that Bt2cAMP similarly increased the PMA-induced transcription from both of these reporter constructs. Taken together, these results suggest that cAMP increases the PMA-induced AP-1 activity which then leads to increased IL-1 beta expression.
Cyclic AMP↗
Tyrosine kinase activity is involved in the protein kinase C induced expression of interleukin-1 beta gene in monocytic cells.
The role of protein tyrosine kinases in the expression of interleukin-1 beta (IL-1 beta) gene in response to phorbol esters (PMA) in THP-1 cell line was investigated. Genistein, a specific tyrosine kinase inhibitor, inhibited PMA induced IL-1 beta protein and mRNA levels in THP-1 cells. Genistein did not have a significant effect on PMA induced activity in transient transfection assays using reporter gene constructs containing the PMA responsive sequence of the IL-1 beta gene linked to IL-1 beta promoter, or five repeats of PMA responsive sites (AP-1 sites) in front of a thymidine kinase promoter. This indicates that the tyrosine kinase activity required for the PMA induced IL-1 beta expression is coupled downstream from or functions independent of the PMA induced AP-1 activity.
Cell Line↗
Continuous presence of phorbol ester is required for its IL-1 beta mRNA stabilizing effect.
The protein kinase C (PKC) activating phorbol esters are known to prevent the decay of mRNA of several cytokines and proto-oncogenes. To examine whether the phorbol ester signal is continuously required for this stabilizing effect, THP-1 monocytic cells were stimulated either with phorbol 12,13-dibutyrate (PDBu), which can be removed from the cells by washings, or with the more hydrophobic phorbol 12-myristate 13-acetate (PMA). Both of these stimuli induced high levels of interleukin-1 beta (IL-1 beta) mRNA. When the cells were washed at the peak of the IL-1 beta mRNA expression, this mRNA decayed rapidly in the PDBu stimulated cells while in PMA stimulated cells the mRNA levels were not affected. Moreover, this mRNA degradation induced by the removal of PDBu could be inhibited by readdition of the phorbol ester. This restabilization could be prevented by pharmacologic inhibitors of PKC, but not by inhibiting protein or RNA synthesis. Thus these data suggest that the phorbol ester must be continuously present to exert its mRNA stabilizing effect and that its effect is PKC-mediated but does not require active protein or RNA synthesis.
Cells, Cultured↗
Signal transduction mechanisms of HLA-DR-mediated interleukin-1 beta production in human monocytes. Role of protein kinase C and tyrosine kinase activation.
The signal transduction pathways leading to the expression of IL-1 beta in human monocytes via HLA-DR stimulation were investigated. SEB, a staphylococcal enterotoxin that binds to HLA-DR molecules, induced IL-1 beta expression in human monocytes. Protein synthesis inhibition by cycloheximide did not inhibit SEB-mediated IL-1 beta signal, indicating that protein synthesis is not required for the MHC class-II-mediated IL-1 beta expression. The effect of PKC, PKA, and tyrosine kinase inhibitors on HLA-DR-mediated IL-1 beta mRNA expression was then determined. H7, a preferential PKC inhibitor, completely inhibited IL-1 beta signal induced by SEB. The role of PKC on HLA-DR-mediated IL-1 beta induction was further confirmed by the ability of SEB to activate PKC on monocytes directly when measured with labeled phorbol ester ([3H]Pbt2)-binding capacity of whole cells. HA 1004, a preferential PKA inhibitor, and isobutyl-methyl-xanthine (IBMX), which inhibits the degradation of cAMP, had no effect on SEB-induced IL-1 beta signal, excluding the role of cAMP on HLA-DR-mediated IL-1 beta expression. Two tyrosine kinase inhibitors, genistein and dihydroxycinnamate, both inhibited SEB-induced IL-1 beta mRNA in monocytes. SEB also induced enhanced tyrosine phosphorylation of several proteins in human monocytes when determined with antiphosphotyrosine immunoblotting. Our results demonstrate that both PKC and protein tyrosine kinases are involved in HLA-DR-induced IL-1 beta expression in human monocytes.
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
Scoring of patellofemoral disorders.
A new questionnaire was used to evaluate subjective symptoms and functional limitations in patellofemoral disorders. The questionnaire was completed independently by four groups of female subjects: controls (N = 17), and subjects with anterior knee pain (N = 16), patellar subluxation (N = 16), and patellar dislocation (N = 19). The questionnaire mean scores for the groups were 100, 83, 68, and 62 points, respectively (p < 0.0001). The items dealing with abnormal painful patellar movements (subluxations) (p < 0.0001), limp (p < 0.0001), pain (p < 0.0001), running (p < 0.0001), climbing stairs (p < 0.0001), and prolonged sitting with the knees flexed (p < 0.0001) differentiated the study groups most clearly. We recommend that these questions be asked when taking a standardized clinical history of an anterior knee pain patient. We also analyzed lateral patellar tilt and displacement by magnetic resonance imaging (MRI) in 28 subjects with patellar subluxation or dislocation. Low questionnaire sum score correlated best with increased lateral patellar tilt measured during quadriceps contraction in 0 degree knee flexion. It seems that a tendency to lateral patellar tilt during quadriceps contraction causes anterior knee pain and can be imaged in knee extension when the patella is not fully supported by femoral condyles.
Cartilage Diseases↗
The lumbar multifidus muscle five years after surgery for a lumbar intervertebral disc herniation.
Biopsy specimens of the lumbar multifidus were obtained from 18 patients with lumbar disc herniation at operation and after a postoperative follow-up period of 5 years. The structure and morphometry of the muscle fibers were analyzed and these data were compared with intraoperative biopsy results and the clinical outcome of the operation. The main findings were: 1) on the basis of occupational handicap score 10 patients belonged in the "positive" and 8 in the "negative" outcome group; 2) the intraoperatively recorded selective type 2 muscle fiber atrophy and the extent of pathologic inner structure changes both decreased in the "positive" outcome group, whereas they persisted in the "negative" group; 3) grouping as a definite sign of reinnervation was seen in only two versus four patients of the "positive" versus "negative" outcome group; 4) the relative amount of adipose tissue within the muscle decreased more markedly in the "positive" outcome group. The authors propose that both inactivity and axonal injury (mainly of neurapraxia type) contribute to the selective type 2 atrophy and inner structure changes in disc patients' multifidus muscle. These pathologic structural changes correlated well with the clinical outcome, and most importantly they are reversible and can be diminished by adequate therapy.
Adult↗
Regulation of endothelial adhesion molecules by ligands binding to the scavenger receptor.
Monocyte adherence to the endothelium, their penetration to the subendothelial space and excessive lipid accumulation (foam cell formation) are the initial events in atherogenesis. Scavenger receptors have been reported to play an important role in foam cell formation, since modified low density lipoproteins can be taken up via scavenger receptors in a non-down-regulated fashion. In this study we demonstrate that stimulation of scavenger receptors in endothelial cells induces the expression of endothelial adhesion molecules. Polyinosinic acid (poly I), a known scavenger receptor ligand, significantly induced the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin on human umbilical vein endothelial cells when compared with polycytidylic acid (poly C), a structurally related compound to poly I, which does not bind to the scavenger receptor. The effect of scavenger receptor ligands on the endothelial cell line EA hy. 926 was also tested. Poly I up-regulated ICAM-1 expression also on EA hy. 926 cells, while it had no effect on IL-1 beta or tumour necrosis factor-alpha (TNF-alpha) production on the same cell line. Poly I-induced ICAM-1 expression on EA hy. 926 cells could be inhibited by H7, a protein kinase C inhibitor, while HA 1004, a preferential protein kinase A inhibitor, had no effect on ICAM-1 expression. The role of protein kinase C in scavenger receptor-mediated adhesion molecule upregulation was confirmed by the ability of poly I to directly activate protein kinase C, when measured with 3H-phorbol dibutyrate binding to EA hy. 926 cells, while poly C again was ineffective.
Cell Adhesion Molecules↗
Okadaic acid, a phosphatase inhibitor, enhances the phorbol ester-induced interleukin-1 beta expression via an AP-1-mediated mechanism.
Protein kinase C (PKC)-activating phorbol esters are known to induce the expression of several genes in monocytic cells. As the effect of serine-threonine kinases, such as PKC, is often counteracted by specific protein phosphatases, we have now examined the role of phosphatases in the regulation of the phorbol ester (PMA)-induced interleukin-1 beta (IL-1 beta) gene expression in the THP-1 monocytic leukaemia cell line. Okadaic acid (OA) is a potent tumour promoter, the function of which is based on its activity to inhibit the serine/threonine specific phosphatases 1 and 2A (PP1 and PP2A, respectively). Thus, it mimicks or potentiates the action of PKC activators in several cell types. Our data demonstrate that alone OA induced a very weak expression of IL-1 beta mRNA, but it strongly enhanced the PMA-induced IL-1 beta expression. To analyse the site of action of OA, the cells were transiently transfected with a chloramphenicol acetyl transferase (CAT)-reporter plasmid containing the AP-1 binding site as the enhancer. Alone, OA was a weak inducer of CAT-activity in these cells, but again it strongly enhanced the PMA-induced response. Similar data were obtained with cells transfected with a reporter plasmid containing the PMA-responsive element (containing a putative AP-1 binding site) of the IL-1 beta gene. Thus, these data indicate that the PMA-induced AP-1 enhancer activity, which is required for the expression of the IL-1 beta gene, is controlled in these cells by PP1 and/or PP2A. As OA did not synergize with PMA in the induction of expression of genes encoding the AP-1 proteins (c-fos, c-jun, junB), it is likely that OA potentiates the AP-1 enhancer activity by its effect on protein phosphorylation.
Cell Line↗
Activin A/erythroid differentiation factor is induced during human monocyte activation.
Activin A/erythroid differentiation factor (EDF), a dimeric polypeptide hormone composed of two beta A subunits, regulates growth and erythroid differentiation of human hematopoietic progenitor and erythroleukemia cells. We have identified activated human peripheral blood monocytes as a natural source of activin A/EDF. In these cells, lipopolysaccharide (LPS) induced rapidly the expression of the beta A subunit mRNAs through protein kinase C-dependent transcriptional regulation. The beta A subunit mRNA expression was also increased by 1,25-dihydroxyvitamin D3, an inducer of macrophage maturation of monocytes. Western analysis with an anti-beta A antibody and an erythroid differentiation bioassay confirmed that the conditioned media of LPS-activated monocytes contained the activin A/EDF protein. We suggest that monocyte/macrophage-derived activin A/EDF may not only modulate hematopoiesis but may also act as a mediator molecule in the diverse physiologic and pathogenetic events in which these cells are involved.
Activins↗
Measles virus infection enhances IL-1 beta but reduces tumor necrosis factor-alpha expression in human monocytes.
Monocytes may play a role in the immunologic abnormalities caused by measles. The effect of measles virus (MV) infection on peripheral blood monocyte functions is poorly known. We report that MV-infected PBM have an altered pattern of IL-1 beta and TNF-alpha production in response to stimulation with LPS and PMA in vitro. MV-infected peripheral blood monocytes produced higher amounts of IL-1 beta, whereas the production of TNF-alpha was reduced. The same effect was observed in the human monocytic cell line THP-1, which was used for RNA analysis. An increased steady-state level of IL-1 beta mRNA was observed in MV-infected cells, and the level of TNF-alpha mRNA was reduced. However, both IL-1 beta and TNF-alpha had about 50% increased transcription rate. Analysis of the mRNA stability after transcriptional block by actinomycin D showed that the TNF-alpha mRNA had a reduced half-life in MV-infected cells (about 30 vs 80 min in uninfected cells), whereas IL-1 beta mRNA stability was similar in uninfected and MV-infected cells. These results indicate that MV infection disturbs the immunoregulatory network by interfering with the monocyte functions.
Base Sequence↗
ABO blood groups and musculoskeletal injuries.
The distribution of the ABO blood groups was studied in 917 patients with specific musculoskeletal diagnoses. The ABO blood group distribution of patients with rupture of the Achilles tendon (P = 0.030) and of patients with chronic Achilles peritendinitis (P = 0.10) differed from the controls. The ABO blood group distribution was not associated with other musculoskeletal injuries studied. The blood group A/O ratio was 1.42 in the control population. In the group with rupture of the Achilles tendon this ratio was 1.0, and in the group with Achilles peritendinitis it was 0.70. The association between injuries of the Achilles tendon and the ABO blood group distribution was in accordance with an earlier report. There may be a genetic linkage between the ABO blood groups and the molecular structure of the tissue of Achilles tendons.
Achilles Tendon↗
Signal transduction pathways leading to the production of IL-8 by human monocytes are differentially regulated by dexamethasone.
Previous studies have shown that IL-8 gene expression is enhanced by various stimuli, which induce different signal transduction pathways. A lipopolysaccharide (LPS)-induced pathway has been reported to be inhibited by glucocorticoids in monocytes. We have now examined the effect of dexamethasone on the LPS-induced and other signal transduction pathways leading to the production of IL-8 by human monocytes. Dexamethasone inhibited the production of IL-8 stimulated with a cyclic adenosine monophosphate analog or LPS. In contrast, dexamethasone had no significant effect on a phorbol ester (PMA)-stimulated IL-8 production. These results suggest that the signal transduction pathways leading to the production of IL-8 by human monocytes are differentially regulated by dexamethasone.
Bucladesine↗
Prolonged elevation of intracellular cyclic AMP activates interleukin-1 production in human peripheral blood monocytes.
The capability of elevated intracellular cyclic AMP concentration to activate IL-1 gene expression and protein production was examined in human peripheral blood monocytes. In accordance with previous studies it was observed that the transiently elevated cyclic AMP (induced either with prostaglandin E2 or with the direct adenylate cyclase activator, forskolin) was not a sufficient signal to activate IL-1 production. However, if the degradation of cyclic AMP was inhibited with isobutyl-methyl-xanthine (IBMX), IL-1 production was strongly activated. This prostaglandin E2 plus IBMX effect could also be mimicked with high concentrations of the cell permeant structural cyclic AMP analogue, dibutyryl cyclic AMP. The cyclic AMP-induced IL-1 production differed in some aspects from the bacterial lipopolysaccharide-induced IL-1 production: (1) the kinetics of both IL-1 gene expression and protein production was much slower; (2) the IL-1 beta gene expression was superinducible by inhibiting the protein synthesis with cycloheximide. Thus these data suggest that prolonged elevation of cyclic AMP is alone a sufficient signal to activate IL-1 production.
Blotting, Northern↗
The effect of calcium mobilization on LPS-induced IL-1 beta production depends on the differentiation stage of the monocytes/macrophages.
The role of elevated intracellular calcium concentration [Ca2+]i in the LPS-induced activation of interleukin-1 beta (IL-1 beta) production was examined in cells representing different stages of myeloid differentiation (undifferentiated monocytic leukaemia cell line THP-1, THP-1 cells induced to adherent, macrophage-like cells by phorbol ester treatment and normal peripheral blood-derived adherent monocytes). LPS did not elevate the [Ca2+]i as measured by the Fura-2 fluorescence technique. When these cells were stimulated with LPS in the presence of the calcium ionophore A23187, a clear increase in the IL-1 beta protein production was observed in the undifferentiated THP-1 cells but not in the more differentiated cell types. This ionophore-induced increase was also seen in the IL-1 beta mRNA levels. Thus these data confirm the previous findings demonstrating that elevation of [Ca2+]i is not involved in the LPS-dependent signal transmission. However, the LPS-induced signals are greatly potentiated by the elevated [Ca2+]i, but only in undifferentiated monocytic cells.
Calcimycin↗