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O Stendahl

Publications and source records attributed to O Stendahl.

At least 73 records · Page 4Linked to original sources
PubMed1988

Does protein kinase C control receptor-mediated phagocytosis in human neutrophils?

It was previously demonstrated that C3bi- but not IgG-mediated phagocytosis in human neutrophils is a Ca2+-independent process [(1985) Nature 315, 509-511]. The objective of this study was to elucidate the nature of an additional messenger signal(s) that may be involved in receptor-mediated phagocytosis in these cells. The C3bi- and IgG-mediated phagocytic ability of neutrophils was inhibited by forskolin, a potent activator of adenylate cyclase. It was found that forskolin induced a 3-fold increase in cAMP levels and simultaneously reduced the uptake of both C3bi- and IgG-opsonized particles by 65%. This inhibition was reversed by exposure to either phorbol myristate acetate (PMA) or diacylglycerol (OAG), which are both activators of protein kinase C, but not by the inactive analog 4 alpha-PMA. PMA could also restore the phagocytic ability of neutrophils with an experimentally impaired calcium response. These results suggest that activation of protein kinase C might be an important transducing signal controlling receptor-mediated phagocytosis in human neutrophils.

Cyclic AMP↗
PubMed1988

Exudate polymorphonuclear leukocytes isolated from skin chambers are primed for enhanced response to subsequent stimulation with chemoattractant f-Met-Leu-Phe and C3-opsonized yeast particles.

The ability to respond metabolically to stimulation with both soluble and particulate substances was investigated in human polymorphonuclear leukocytes (PMNLs) isolated from an aseptic inflammatory reaction. Exudate PMNLs isolated from skin chambers (E-PMNLs) and blood PMNLs isolated from the peripheral blood (B-PMNLs) of the same individual were investigated in parallel. E-PMNLs were primed, resulting in an increased chemiluminescence (CL) response to subsequent stimulation with the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) (334%) and serum-opsonized yeast particles (C3 yeast) (201%), as compared to B-PMNLs. Phorbol myristate acetate (PMA) on the other hand, induced a CL response in E-PMNLs that was only 70% of the response obtained in B-PMNLs. A similar primed state resulting in enhancement of the CL response to FMLP and C3 yeast could be induced in B-PMNLs by pretreatment with a bacterial culture filtrate. Pretreatment of E-PMNLs with the bacterial culture filtrate, however, did not increase the CL response to FMLPs any further. The enhanced functional response to FMLPs in E-PMNLs was accompanied by an increased binding of the peptide, demonstrated by a doubling of the amount of bound f-Met-Leu-[3H]Phe (209%), as compared to B-PMNLs. The increased C3-yeast-induced CL generation in E-PMNLs was accompanied by an increased ingestion and attachment of C3-opsonized yeast particles. The enhancement of phagocytosis in E-PMNLs was, however, dependent upon the opsonin used, since IgG-opsonized yeast particles were phagocytosed to the same extent by E-PMNLs and B-PMNLs, thereby indicating that selective receptor modulation is also involved in the priming of E-PMNLs for an enhanced response to C3-yeast. These results show that exudate cells isolated from skin chambers are modulated with respect to receptor-mediated functions resulting in an increased metabolic response to FMLP coupled with an increased binding of the peptide and an increased phagocytosis of C3-coated yeast particles. Receptor modulation during exudation may be an important mechanism in regulating the inflammatory response by PMNLs.

Chemotaxis, Leukocyte↗
PubMed1988

Reduced redox potential during growth of some gram-negative bacteria. Effect on the biochemical and physicochemical surface properties and phagocytosis by polymorphonuclear leukocytes.

When cultivated at reduced redox potential the physico-chemical surface properties were altered in strains of E. coli, Salmonella and Yersinia bacteria. In particular, strains which showed hydrophilic surface properties under normal aerobic cultivation became more hydrophobic when exposed to anaerobic conditions (e.g. E. coli K12, E. coli K12D21, E. coli K12D22, S. minnesota S99, S. typhimurium 395MS, S. braenderup 2828 and Yersinia enterocolitica). Moreover, there were qualitative as well as quantitative differences in the protein profiles of whole bacterial lysates and membrane preparations analysed in SDS-PAGE. There were no qualitative differences in the lipopolysaccharide (LPS) bands. However, when E. coli K12D22 were cultivated aerobically, remarkably more high molecular temperature-sensitive (70 degrees C for 45 min) carbohydrate material was produced (weight about 360 KD and 660 KD). Interaction between polymorphonuclear leukocytes (PMNL) and the E. coli K12D22 strain, measured as chemiluminescence, showed that the anaerobically cultivated bacteria induced a chemiluminescence that was mainly of intracellular origin, while the aerobically cultivated induced an extracellular response. Phagocytosis and killing-studies showed that only anaerobically-grown E. coli were effectively inactivated by the PMNL.

Aerobiosis↗
PubMed1987

Chemotactic factor binding and functional capacity: a comparison between human granulocytes and differentiated HL-60 cells.

In the presence of dimethyl sulfoxide (DMSO), the leukemic promyelocytic cell line HL-60 will differentiate into mature polymorphonuclear granulocytes. In the present report, we compare chemotactic factor binding and function in HL-60 cells with that of normal human granulocytes using the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (FMLP) as ligand. The cellular response measured as CL was changed as a result of storage or conditioning of normal peripheral blood cells. With these cells, a conditioning procedure at room temperature resulted in a pronounced increase in the CL response. The increase of the CL response was probably a result of increased expression of cryptic receptors, since the changes of the oxidative response to FMLP was accompanied by increased binding of the peptide to the cell surface. Scatchard analysis revealed that the increased binding was due to an increased number of receptors. In differentiated HL-60 cells, conditioning neither led to increased production of oxidative metabolites, nor to any increased binding of the peptide. The data thus indicate that many FMLP receptors could reside in a cryptic site that is not accessible to extracellular ligands, and that conditioning results in an increased exposure of these receptors, followed by an increased oxidative response to the ligand in normal cells but not in mature HL-60 cells.

Cell Line↗
PubMed1987

Cell surface expression of fMet-Leu-Phe receptors on human neutrophils. Correlation to changes in the cytosolic free Ca2+ level and action of phorbol myristate acetate.

We have studied how cytosolic free Ca2+ ([Ca2+]i) changes and phorbol myristate acetate (PMA) exposure affects ligand-independent cell surface expression of fMet-Leu-Phe receptors on human neutrophils. Mere incubation primed neutrophils to double their binding of fMet-Leu-Phe. This spontaneous increase of peptide binding was unaffected by changes in the extracellular calcium concentration. However, depression of the [Ca2+]i totally abolished the increased binding of fMet-Leu-Phe. Scatchard-Plot analysis revealed that the observed increase of peptide binding was due to an increased number of receptors. Normalization of the [Ca2+]i in cells where it was initially depressed resulted in a slow but progressive increase in fMet-Leu-Phe binding. The rate of receptor recruitment could be enhanced by rapidly increasing the [Ca2+]i by addition of ionomycin. Addition of PMA to cells with near maximal receptor expression led to a marked reduction of fMet-Leu-Phe binding without affecting [Ca2+]i. These observations suggest the existence of a dual regulatory mechanism for up- and down-regulation of fMet-Leu-Phe receptors on the cell surface of human neutrophils.

Aminoquinolines↗
PubMed1987

Neutrophil function in psoriasis: effects of retinoids.

The present investigation focused on the oxidative response of polymorphonuclear neutrophil leukocytes in psoriasis, in particular pustular psoriasis and how this response was affected by different retinoid compounds. In the active phase of pustular psoriasis, the neutrophil chemiluminescence response to the chemotactic peptide f-met-leu-phe and to phorbol myristate acetate was enhanced and correlated to the development of pustules, whereas cells from psoriasis vulgaris patients showed normal chemiluminescence response. Retinoids, particularly tretinoin (= retinoic acid) and isotretinoin caused a pronounced inhibition of the chemiluminescence response only in primed neutrophils in vivo and in vitro, whereas etretinate and the metabolite Ro 10-1670 was less inhibitory. Retinoic acid furthermore inhibited the Fc-mediated phagocytosis, but did not affect C3bi-mediated phagocytosis. These data suggest that the antiinflammatory effect of retinoids may operate by affecting neutrophil activation and function.

Acitretin↗
PubMed1986

Leukotriene B4 stimulation of phagocytes results in the formation of inositol 1,4,5-trisphosphate. A second messenger for Ca2+ mobilization.

Inositol trisphosphate (InsP3) production and cytosolic free Ca2+ ([Ca2+]i) elevations induced by leukotriene B4 (LTB4)-receptor activation were studied in the human promyelocytic-leukaemia cell line HL60, induced to differentiate by retinoic acid. The myeloid-differentiated HL60 cells respond to LTB4 by raising their [Ca2+]i with a dose-response relationship similar to that shown by normal human neutrophils. The observations of the LTB4 transduction mechanism were compared with those of the transduction mechanism of the chemotactic peptide fMet-Leu-Phe in HL60 cells differentiated with dimethyl sulphoxide. Both LTB4 and fMet-Leu-Phe triggered a rapid (less than 5 s) elevation of [Ca2+]i, which occurred in parallel with the InsP3 production from myo-[3H]inositol-labelled cells. The threshold concentrations of the agonists, for InsP3 production, were found at 10(-9) M, a slightly higher concentration than that required to detect [Ca2+]i elevations. No significant changes were noted in the phosphoinositide levels upon stimulation with LTB4. Exposure to Bordetella pertussis toxin before LTB4 stimulation abolished both the increased formation of InsP3 and the rise of [Ca2+]i. LTB4 and fMet-Leu-Phe elicited elevations of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] with no detectable lag time, followed by slower and more sustained inositol 1,3,4-trisphosphate elevations. Stimulation with various leukotriene analogues revealed a good correlation between both total InsP3 as well as Ins(1,4,5)P3 formation and elevations of [Ca2+]1. Thus LTB4 receptor activation results in an increased production of Ins(1,4,5)P3 via a transduction mechanism also involving a nucleotide regulatory protein, as previously described for the fMet-Leu-Phe transduction mechanism.

Calcium↗
PubMed1986

Defective chemiluminescence response in differentiated HL60 cells due to impaired degranulation.

In the presence of dimethyl sulfoxide, the promyelocytic leukemic cell line, HL60, differentiates into apparently mature polymorphonuclear leukocytes. When correlating the superoxide production from HL60 cells with the number of phagocytozing and NBT-positive cells, no difference was observed in comparison with normal peripheral blood leukocytes. In contrast, the luminol-dependent chemiluminescence was greatly impaired in the differentiated HL60 cells. Analysis of degranulation, i.e., release of myeloperoxidase and N-acetyl-beta-glucosaminidase- and myeloperoxidase-mediated iodination by HL60 cells, suggested that the defective chemiluminescence response observed in HL60 cells may be due to impaired release of myeloperoxidase from azurophilic granulae. This may lead to impaired microbicidal activity in these cells.

Cell Differentiation↗
PubMed1986

The kinetics of peritoneal clearance of Escherichia coli and Bacteroides fragilis and participating defense mechanisms.

In a study using pigs, we delineated the dynamic character and the interplay of defense mechanisms that operate in the peritoneal cavity and their effectiveness against Escherichia coli and Bacteroides fragilis. The bacteria were extensively cleared during the first three hours. The concentration of both organisms in the peritoneal fluid decreased by up to 300,000-fold, and the final concentration was a function of the inoculum given. Clearance efficiency did not differ between a bolus dose and prolonged bacterial administration. Peritoneal absorption was a major eliminating mechanism during the first three hours but was limited to bacterial concentrations exceeding approximately 10(9) colony-forming units per milliliter of peritoneal fluid. Intraperitoneal elimination started immediately and continued for approximately six hours. Thereafter, residual bacteria were not eliminated, even though the defense capacity was intact. Prolonged bacterial administration, however, protracted the period for active bacterial destruction.

Animals↗
PubMed1986

Bacterial clearance and granulocyte response in experimental peritonitis.

Clearance kinetics of Escherichia coli and Bacteroides fragilis from the peritoneal cavity and functional properties of peritoneal and blood polymorphonuclear leukocytes (PMNL) were studied in pigs. The bacteria, approximately 10(10) colony forming units (CFU) of each species, were intraperitoneally injected. Blood and peritoneal exudate were sampled regularly during 24 hr for bacterial quantification and PMNL analysis. The peritoneal concentration of both species fell rapidly in the first 3 hr and then stabilized. Approximately 10(2) CFU/ml remained after 24 hr. Chemotaxis, phagocytosis, and metabolic activation of the peritoneal PMNL showed little change. In some pigs a second bacterial dose was injected 6 hr after the first dose and gave an identical clearance pattern. Exhaustion of the local defence capacity thus was excluded as cause of the impaired bacterial elimination.

Animals↗
PubMed1986

Superoxide production and chemiluminescence induced in differentiated HL-60 cells by the chemoattractant formyl-methionyl-leucyl-phenylalanine.

Superoxide production and chemiluminescence induced in differentiated HL-60 cells by the chemoattractant formylmethionyl-leucyl-phenylalanine: In order to study the generation of oxidative metabolites in relation to cell differentiation, dimethyl sulfoxide (DMSO) and retinoic acid (RA) differentiated HL-60 cells were stimulated with the chemotactic peptide formylmethionyl-leucyl-phenylalanine (FMLP). The oxidative response was measured as luminol-dependent chemiluminescence, lucigenin-dependent chemiluminescence, and cytochrome c reduction. Cells grown in the presence of DMSO or RA progressively expressed morphological changes, and when the mature cells were exposed to FMLP the cells produced oxidative metabolites. Quantitatively the HL-60 cells grown in the presence of DMSO gave rise to the most pronounced response. No correlation was obtained between superoxide production, luminol-chemiluminescence and lucigenin-dependent chemiluminescence, indicating that different aspects of the oxidative response are elucidated by the three different methods. Furthermore, the experiments show that DMSO and RA-induced differentiation of HL-60 cells leads to granulocyte-like cells with different abilities to produce oxidative metabolites, possibly due to differences in receptor function.

Cell Differentiation↗
PubMed1986

Factor-specific deactivation of leucocyte chemotaxis in vivo.

This report analyses the locomotory capacity of polymorphonuclear leucocytes (PMNL) isolated from 7 patients with bacterial meningitis. 7 healthy control subjects were also investigated in parallel. It was found that PMNL from the patients suffering from meningitis, isolated both from peripheral blood and from the cerebrospinal fluid, had lost their ability to respond chemotactically to activated serum but not to the chemotactic peptide f-Met-Leu-Phe. The normal chemotactic responsiveness of blood PMNL was restored once the patients recovered from infection.

Blood Physiological Phenomena↗
PubMed1986

Characterization of fMet-Leu-Phe receptor-mediated Ca2+ influx across the plasma membrane of human neutrophils.

N-Formyl-L-methionyl-L-leucyl-L-phenylalanine (fMet-Leu-Phe) stimulation of human neutrophils leads to a rapid increase of the cytosolic free Ca2+ concentration, [Ca2+]i, which is significantly reduced by removal of extracellular calcium. In the present study we show that fMet-Leu-Phe-induced [Ca2+]i increases are, in part, mediated by an increase of the plasma membrane permeability to Ca2+. This conclusion is based on the following evidence. In the presence of extracellular calcium, addition of La3+ reduced the fMet-Leu-Phe-induced [Ca2+]i increase to approximately the same level as that observed in the absence of extracellular calcium. A net increase of the plasma membrane permeability for Mn2+ could be observed after fMet-Leu-Phe stimulation, as revealed by intracellular quenching of the quin2 signal. The influx of Mn2+, like that of Ca2+, was inhibited by La3+ and was more pronounced in the absence of extracellular Ca2+, suggesting competition for the same pathway. Temporal dissociation of intracellular Ca2+ release from stores and Ca2+ influx from the medium could be demonstrated by readdition of calcium to cells stimulated in the absence of this cation. This second [Ca2+]i increase could be abolished either by giving the specific chemotactic peptide receptor antagonist, BOC-Met-Leu-Phe, or Co2+. We could also show that the fMet-Leu-Phe-dependent Ca2+ influx was not due to the activation of voltage-dependent calcium channels since depolarization either by K+ or gramicidin D did not affect the resting [Ca2+]i, nor did it affect a subsequent [Ca2+]i increase induced by fMet-Leu-Phe. Furthermore, nifedipine and verapamil, at concentrations known to block classical voltage-dependent calcium channels, had no significant effects on the Ca2+ influx induced by fMet-Leu-Phe. We suggest that fMet-Leu-Phe promotes influx of Ca2+ ions across the plasma membrane of human neutrophils by opening of receptor-dependent calcium channels.

Adult↗
PubMed1986

The role of the cytosolic free Ca2+ transient for fMet-Leu-Phe induced actin polymerization in human neutrophils.

We have addressed the important question as to if and how the cytosolic free Ca2+ concentration, [Ca2+]i, is involved in fMet-Leu-Phe induced actin polymerization in human neutrophils. Stimulation of human neutrophils with the chemotactic peptide (10(-7) M), known to result in a prompt rise of the [Ca2+]i to above 500 nM, also induced a rapid decrease of monomeric actin, G-actin, content (to 35% of basal) and increase of filamentous actin, F-actin, content (to 320% of basal). A reduction of the fMet-Leu-Phe induced [Ca2+]i transient to about 250 nM, resulted in a less pronounced decrease of G-actin content (to 80% of basal) and increase of F-actin content (to 235% of basal). A total abolishment of the chemotactic peptide induced [Ca2+]i rise, still led to a decrease of the G-actin content (to 85% of basal) and increase of F-actin (to 200% of basal). These results indicate that the [Ca2+]i rise is not an absolute requirement, but has a modulating role for the fMet-Leu-Phe induced actin polymerization. Another possible intracellular candidate for fMet-Leu-Phe induced actin polymerization is protein kinase C. However, direct activation of protein kinase C by phorbol 12-myristate 13-acetate (PMA) only resulted in a minor increase of F-actin content. The recent hypothesis that a metabolite of the polyphosphoinositide cycle, independently of [Ca2+]i and protein kinase C, is responsible for actin polymerization agrees well with these results and by the fact that preexposure to pertussis toxin totally abolished a subsequent increase of F-actin content induced by fMet-Leu-Phe.

Actin Cytoskeleton↗
PubMed1985

Inhibition of human leukocyte metabolism and random mobility by local anaesthesia.

Hexose-monophosphate shunt (HMS) activity, myelo-peroxidase-(MPO)-mediated iodination and random mobility in human polymorphonuclear leukocytes (PMNs) were studied in the presence of lignocaine. Incubating the PMNs with 0.1% lignocaine during phagocytosis inhibited the 14CO2 produced from glucose-1-14-C via the HMS shunt by 33%. On increasing the concentration of lignocaine, a dose-dependent inhibition was noted. The MPO-mediated iodination was inhibited by 73% in the presence of 0.1% lignocaine, and complete inhibition took place when the concentration was increased to 0.5%. The random mobility of leukocytes was studied by an opto-electronic technique. In the presence of 0.5% lignocaine, all leukocytes examined were completely immobilized; in the presence of 0.1% lignocaine immobilization took place within 45-65 min.

Anesthetics, Local↗
PubMed1985

Mannose-specific and hydrophobic interaction between Escherichia coli and polymorphonuclear leukocytes--influence of bacterial culture period.

The influence of culture period on mannose-specific and hydrophobic properties of the bacterial surface and on bacteria/polymorphonuclear leukocyte (PMNL) interaction was studied. Four E. coli strains, PN7 (01:K1), ABU2 (ON:K14), CU9 (06:K14) and CU13 (08:KN) and two Salmonella typhimurium strains 395 MR10 and 395 MS, well characterized according to physicochemical surface properties, presence of type 1 fimbriae and interaction with PMNL, were used in the study. The results show that with prolonged culture period, the liability to hydrophobic interaction increases, the agglutination-strength of mannose-specific maltobionamide liposomes increases, while the agglutination-titer with guinea-pig erythrocytes remains constant. Furthermore, the mannose-specific association with and metabolic activation of PMNL is augmented, while the ingestion is unchanged. In addition, our results demonstrate differences in sensitivity between the methods used to detect exposure of mannose-specific structures on the surface of bacteria, and that the culture condition is important for bacterial surface properties. It thus appears that the culture conditions have a great influence on the surface properties of E. coli bacteria and the interaction with phagocytic cells.

Escherichia coli↗