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Biomedical subjects

U Hadding

Publications and source records attributed to U Hadding.

At least 55 records · Page 3Linked to original sources
PubMed1984

Regulation of the oxidative burst of macrophages by lipid mediators.

Macrophages, on stimulation, generate both cyclo-oxygenase and lipoxygenase products of arachidonic acid as well as the acetylated phospholipid, PAF-acether. The latter induces the oxidative burst in C. parvum-activated macrophages, but has little activity on resident macrophages. Mouse macrophages are generally rather insensitive to PAF-acether. Prostaglandin (PG)E2, on the other hand is a potent inhibitor of the macrophage oxidative burst, though its activity is also dependent upon the source of the macrophages and the stimulus used. Several lipoxygenase inhibitors inhibit the oxidative burst of mouse peritoneal macrophages, while other authors have reported stimulatory effects of lipoxygenase products on different macrophage populations. The differential actions of lipoxygenase and cyclo-oxygenase products on the macrophage oxidative burst are reflected by the effect of arachidonic acid on the zymosan induced response of mouse resident peritoneal cells: initial enhancement, followed by prolonged inhibition, the latter being indomethacin-sensitive. While PAF-acether is probably an important initiator of the macrophage oxidative burst in vivo, arachidonic acid modulates the response by a "see-saw mechanism", lipoxygenase products being stimulatory and cyclo-oxygenase products (mainly PGE2) being inhibitory.

Animals↗
PubMed1984

Expression of Ia antigens on macrophages is reduced after stimulation with homologous C3b.

The complement cleavage product C3b was examined for its ability to influence the expression of Ia antigens on macrophages. Monoclonal anti-Ia antibodies were applied for quantitation of Ia antigens by using FACS analysis. Albumin-elicited guinea pig peritoneal macrophages were cultured for 48 hr in the presence of C3b. C3b at final concentrations varying from 10 to 80 micrograms/ml reduced the proportion of Ia-positive macrophages from 87 to 25%. The C3b-stimulated loss of Ia antigens from the macrophage surface was mediated by metabolites of the cyclooxygenase pathway as shown by the inhibitory effect of indomethacin. Exposure of macrophages to C3b for 5 min was sufficient to induce Ia reduction, seen after subsequent 48-hr incubation. C3b generated in vivo in the course of inflammatory reactions might therefore influence the interaction of immunocompetent cells by modulating macrophage Ia expression.

Animals↗
PubMed1983

Synthesis of complement by macrophages and modulation of their functions through complement activation.

During the last decade considerable progress has been made to characterize intimate functional links between macrophages, a major cellular component of immunoinflammatory responses, and the complement system representing the major humoral mediator of inflammation. Macrophages of various species and tissue sites have been shown to synthesize and release most of the complement components providing these cells with their own "pericellular" complement system. Circumstantial evidence for the assembly of both classical and alternative pathway convertases has been adduced. An intricate network of feedback loops involving endogenous and extrinsic factors operates to adjust complement production to acute requirements, for example augmenting production in the face of accelerated turnover at sites of inflammation, and returning it to baseline levels once the inflammatory stimulus has subsided, in order to maintain a fine-tuned balance. The molecular mechanisms underlying regulation of complement synthesis by macrophages are beginning to be elucidated by use of gene technology. On the other hand, complement activation products exert a number of effects on macrophages via specific surface receptors causing internalization of offending agents, microbes, and immune complexes, promotion of intracellular killing, controlling migration behavior, inducing release of potent biologic substances such as lysosomal enzymes, arachidonic acid metabolites, and interleukin 1. In these interactions, two important humoral mediator systems of inflammation, the complement system and the arachidonic acid cascade, are functionally linked at the level of the macrophage. Stimulation of the release of immunomodulating compounds from macrophages invoke a role for complement in immune regulation. This multifaceted interplay is of particular importance considering the mobility of macrophages that allows them to gain almost unrestricted access to sites of ongoing immunoinflammatory responses. The time seems to have come to abandon the petrified thinking in socalled systems as, for instance, humoral versus cellular, specific versus unspecific, and to proceed to interlocking functions guided by physiology proper.

Anaphylatoxins↗
PubMed1983

Platelet activating factor (PAF) induces the oxidative burst in macrophages.

The response of guinea pig peritoneal macrophages to platelet activating factor (1-O-octadecyl-2-acetyl-sn-glycero-3-phosphoryl-choline) was examined. In Corynebacterium parvum induced macrophages, platelet activating factor, over a wide dose range (3.8 X 10(-5) to 3.8 X 10(-9)M) triggered the oxidative burst as indicated by increased luminol-dependent chemiluminescence and hydrogen peroxide release into culture supernatants. This effect of PAF was inhibited by superoxide dismutase and catalase. Resident macrophages exhibited only slight respiratory activity in response to platelet activating factor which could be increased by adding 1% gelatine to the medium. Activation of macrophages is a new biological effect exerted by platelet activating factor. In immuno-inflammatory reactions, cells capable of generating platelet activating factor may come into close contact with macrophages and by liberating this mediator cause them to release highly toxic oxygen species known to be microbicidal and cytocidal and able to produce vascular endothelial injury. Our findings lend further support to the view that platelet activating factor is a potent and rapid activator of physiological defence mechanisms.

Animals↗
PubMed1983

Stimulation of prostaglandin E and thromboxane synthesis in macrophages by purified C3b.

Complement cleavage product C3b was examined for its effect on macrophages. Albumin-elicited guinea pig peritoneal macrophages kept in short-term culture were challenged with purified homologous C3b, and release of oxygen and arachidonic acid metabolites was studied. C3b at concentrations ranging from 20 to 80 micrograms/ml induced synthesis of large quantities of the arachidonic acid cyclooxygenation products thromboxane B2 (TXB2) and prostaglandin E (PGE) but failed to trigger an oxidative burst. Our findings link together macrophages, complement, and arachidonate metabolites as major participants in inflammatory responses.

Animals↗
PubMed1983

Induction of thromboxane release from macrophages by anaphylatoxic peptide C3a of complement and synthetic hexapeptide C3a 72-77.

Guinea pig peritoneal macrophages were examined for their response to homologous anaphylatoxic peptide C3a of complement. C3a caused the release of considerable amounts of thromboxane A2 from macrophages maintained in serum-free culture for up to 12 hr. Preincubation of C3a with the Fab portion of a monoclonal anti-C3a IgG abrogated its stimulatory capacity. C3a desarg was found to be rather ineffective in eliciting thromboxane release. Hexapeptide C3a 72-77, comprising the C-terminal amino acids of human C3a, proved also to stimulate the cyclooxygenation pathway of macrophage arachidonic acid metabolism, following the same routes of receptor-ligand interactions (rapid onset of activation, low dose, and cumulative desensitization) as proven for other anaphylatoxin-cell interactions. This novel effect of anaphylatoxins C3a and C3a 72-77 could contribute to inflammatory reactions in which macrophages play a role.

Animals↗
PubMed1982

Low zone desensitization: a stimulus-specific control mechanism of cell response. Investigations on anaphylatoxin-induced platelet secretion.

The biologic activity of the anaphylatoxic peptides C5a and C3a is regulated efficiently at the target-cell level by the phenomenon of desensitization. Desensitization of platelets is stimulus specific and can be induced by low concentrations of anaphylatoxins without any preceding secretory event. In contrast to activation to secretion, desensitization is Ca++ independent but much more time consuming, especially at lower temperatures where both processes differ markedly in reaction velocity. This low zone desensitization insures that secretion from platelets only occurs when high amounts of anaphylatoxins are rapidly generated in the vicinity of the target-cell. Consequently, stimulus-specific unresponsiveness of the target cells can be induced by slowly increasing the concentration of the respective stimuli in their vicinity. Cellular control seems to act as a first-line mechanism of regulation, whereas the role of fluid-phase control is considered as preventing longer persistence and systemic accumulation of active anaphylatoxins.

Anaphylatoxins↗
PubMed1982

Quantitative studies of the secretion of complement component C3 by resident, elicited and activated macrophages. Comparison with C2, C4 and lysosomal enzyme release.

To quantitate the secretion of complement component C3 by guinea pig peritoneal macrophages an enzyme-linked immunosorbent assay was developed. C3 secretion was studied in resident, elicited and activated macrophages and compared with release of hemolytically active C2 and C4, as well as the lysosomal enzyme beta-D-2-acetamido-2-deoxyglucosidase. Resident macrophages secreted about 6 ng C3/10(6) cells/h into culture supernatants over a period of 12 h. Corynebacterium parvum-activated cells were found to secrete 3 times that amount at nearly constant rates. There was a stepwise increase in secretion of functional C2 and C4 when comparing resident, elicited and activated macrophages; secretion was 2--4 times higher in activated than in resident cells.

Acetylglucosaminidase↗
PubMed1982

C3a induced activation and stimulus specific reversible desensitization of guinea pig platelets.

C3a and its C-terminal hexapeptide lead to a dose dependent release of biogenic amines and nucleotides stored in platelet's granules. The release reaction can be measured by tritiated serotonin or by ATP, indicated by an ATP specific bioluminescence assay. We tested the capability of C3a to induce aggregation of washed platelets. The recently described phenomenon of low dose, stimulus specific desensitization of platelets to the anaphylatoxic peptides C3a and C5a could be shown by measuring the release reaction as well as the aggregation. Further we could demonstrate the reversibility of the stimulus specific desensitization within 2 to 3 hours. The desensitization was proven to be temperature dependent. The recovery of function was independent of newly synthetized protein and is discussed as the result of receptor-recycling.

Anaphylatoxins↗
PubMed1981

Activation of the alternative pathway of complement: efficient fluid-phase amplification by blockade of the regulatory complement protein beta1H through sulfated polyanions.

Current concepts of activation of the alternative pathway of complement (APC) focus on the central role of an amplification mechanism triggered by C3b which is covalently bound to the surfact of activating substances. Using sulfated polyanions as model substances, an efficient fluid-phase activation of complement is demonstrated in contrast to solid-phase activation. It is shown that particulate high-molecular weight sulfated polyanions are capable of reversible binding the guinea pig and human regulatory protein beta1H. This fixation leads to an extensive activation of C3 and factor B because the regulatory function of beta1H is blocked in the fluid-phase C3b-dependent amplification system of the APC. Addition of beta1H-depleted C4-deficient guinea pig serum reconstitutes the physiological control mechanisms of the APC. Guinea pig beta1H, purified to homogeneity, is described as a 160000 dalton protein of a single-chain structure. In addition, highly specific and sensitive test systems for beta1H are described.

Absorption↗
PubMed1981

Comparative study on biological activities of various anaphylatoxins (C4a, C3a, C5a). Investigations on their ability to induce platelet secretion.

Several anaphylatoxic substances (human C3a, guinea pig C3a, human C4a, guinea pig C5a, and a synthetic C3a-related hexapeptide) were compared with regard to their ability to induce secretion of [3H] serotonin from guinea pig platelets. Functional identity of the C3a preparations, C4a, and the hexapeptide was demonstrated by the phenomenon of crossed desensitization. Whereas C3a of human and guinea pig origin proved to be qualitatively and quantitatively identical, C4a expressed only 3% of the activity of the C3 fragments on a molar basis. Investigations with goat anti-guinea pig C3a demonstrate that human and guinea pig C3a possess one antigenic determinant in common; however, this determinant is not the C-terminal amino acid sequence. Addition of the anaphylatoxins with low doses of thrombin led to a potentiation of [3H] serotonin release from the platelets. Under these conditions C3a concentrations of 1.5 X 10(-10) mumol/liter (65 pg of C3a) could be detected. Thus the platelet system represents the most sensitive in vitro assay known for evaluation of biological activity of the C3a anaphylatoxins.

Anaphylatoxins↗
PubMed1981

Synthetic C3a analogs as specific inhibitors of C3a activity.

Various C3a-related C-terminal synthetic oligopeptides were investigated for their ability to induce a release of serotonin from guinea pig platelets. The results confirm earlier findings that expression of biological C3a activity requires the four to five C-terminal amino acids of the C3a primary structure and underlines the essential role of the C-terminal arginine. Besides their ability to induce a specific release reaction, these peptides--after short incubation with the platelets--lead to a specific desensitization of the cells for C3a or C3a-related stimuli. Expression of this inhibitory activity required concentrations of the peptides more than 100-fold lower than those that were necessary to induce secretion. The possibility of using C3a analogs as specific inhibitors for C3a offers a valuable tool for in vivo studies of biological C3a activity.

Amino Acid Sequence↗
PubMed1981

Platelet-serotonin release by C3a and C5a: two independent pathways of activation.

C3a and C5a were investigated for their ability to induce secretion of serotonin from platelets in a homologous guinea pig system. Platelets respond to either anaphylatoxin with a dose-dependent release that does not exceed a plateau at the 70% level. On a molar basis. C5a is about 50-fold more active than the C3 fragment. The C5a effects in the platelet system can be clearly distinguished from the activity of C3a and vice versa by a phenomenon of specific desensitization of the platelets to either stimulus. Our observations lead to the postulation of distinct receptors for C3a and C5a on guinea pig platelets as well as 2 independent pathways of platelet activation, which both lead to a specific release.

Animals↗
PubMed1981

Linkage of total deficiency of the second component (C2) of the complement system and of genetic C2-polymorphism to the major histocompatibility complex of the guinea pig.

In the guinea pig six common phenotypes for hemolytically active C2 were detected. They resulted from three allotypic variants: a basic C2B and two more acidic variants C2A and C2A1. (The gene frequency for C2B in the outbred population was 0.36, 0.33 for C2A, and 0.31 for C2A1.) These variants were inherited as autosomal codominant traits. A strong linkage-disequilibrium between C2 and C4 allotypes was observed consisting of the pairs C2A1-C4F. C2A-C4S, whereas C2B was associated with either C4S or C4S1. To investigate the linkage of C2 to the MHC a family of strain 2 X strain OM3 was studied: the C2B allotype segregated with the strain haplotype of OM3 whereas C2A1 does the same with the strain 2 haplotype. During the C2-typing, an animal totally deficient in hemolytic C2 activity was discovered. Its C1, C4, C3, C5, factors B, D, and H were normal. Breeding yielded finally homozygous and heterozygous-deficient individuals. The latter ones exhibited 50 to 60% of the hemolytic activity of normal guinea pigs. Combined typing data of C4, C2 and GPLA showed that the deficient gene behaved like a rare, silent allele of C2, which is GPLA-linked and inherited like the normal C2-variants. An antiserum against C2 was raised in C2-deficient animals, which reacted with normal guinea pig serum but not with serum from C2-deficient animals. No cross-reactivity with human or mouse serum could be detected.

Alleles↗