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The properties of human C5a anaphylatoxin. The significance of C5a formation during hemodialysis.

Human C5a anaphylatoxin is a potent bioactive molecule that possesses both spasmogenic and leukocyte-related properties. As such, it normally serves as a local mediator of the acute inflammatory response. Additionally, C5a, through its actions of mononuclear phagocytes, may act to bridge the gap in the acute-chronic inflammatory continuum. While these properties are critical to normal host defense mechanisms, it is now apparent that this anaphylatoxin and/or its des-Arg74 derivative, may exert significant systemic effects that are manifest as cardiopulmonary abnormalities and intravascular activation of granulocytes. Knowledge of these properties is critically important for understanding the clinical sequelae exhibited by patients undergoing extracorporeal circulation since we now know that both hemodialysis and cardiopulmonary bypass [28-30] procedures promote intravascular complement activation and C5a formation. Viewed in this context, it seems reasonable to postulate that many of the immediate and delayed responses to extracorporeal circulation might be mediated by C5a formed in the extracorporeal circuit (table IV). For example, it is now recognized that a few particularly susceptible patients display adverse reactions during the initial phases of hemodialysis. The symptoms of this so-called 'first-use syndrome' may range from severe urticaria and angioedema to life-threatening bronchospasm, hypotension, and cardiopulmonary collapse. Some investigators have presented data which suggest that complement-derived products may be causative of these symptoms in some patients [31]. While this hypothesis remains to be confirmed, present evidence clearly demonstrates that C5a alone may produce many of the observed phenomena. In addition to the acute effects produced by C5a, both our own basic studies and the clinical investigations presented by others at this conference suggest that the long-term effects of repeated C5a exposure in the dialyzed patient may be considerable. Thus, there has been a great deal of interest in the role of complement-derived mediators as initiators of leukocyte degranulation and toxic oxygen radical production and an exploration of the significance of these events in the eventual development of chronic pulmonary fibrosis in the dialyzed patient. Similarly, the effects of repeated exposure to IL-1 that has been postulated to occur as a result of C5a triggering of monocytes during dialysis is currently an active area of investigation.(ABSTRACT TRUNCATED AT 400 WORDS)

Anaphylatoxins

Suppression of vascular injury at the Arthus reaction site by a complement inhibitor, 5,5',5''-(1,3,6-naphthalene-triyl-tris[sulfonylimino])-tris (1,3-benzene disulfonic acid) hexasodium salt.

A novel polyanionic complement inhibitor 5,5,5''-(1,3,6-naphthalene-triyl-tris[sulfonylimino])-tris(1 ,3-benzene- disulfonic acid) hexasodium salt (compound IIb) was tested for its ability to suppress vascular injury at the site of the Arthus reaction (AR). In control animals in which AR was evoked without drug treatment, venules at AR sites ranged from normal (arbitrarily defined as stage I) to destroyed (stage V). Between these two ends of the spectrum were venules with an accumulation of cells and deposits of electron dense material (stage II), accumulations of cells and deposits and small endothelial gappings (stage III), and accumulations of cells and depositions which had spread into perivascular tissue and small gappings (stage IV). In animals treated with compound IIb, the AR stopped at stage III or IV depending on the dose, it never reached stage V. In other words compound IIb treatment resulted in protection of endothelium, basal lamina and other structures from the destruction which is characteristically observed in the AR. The effect of high doses of compound IIb was similar to that described before for suramin.

Animals

Relationships of serum opsonins and complement in human experimental gingivitis.

This study assessed the phagocytosis of polymorphonuclear neutrophils and the opsonic capacity of serum in experimental human gingivitis. Relationships were sought among these two measurements, clinical indices of gingivitis and complement-related opsonins. Measurements of chemiluminescence provided an index of phagocytosis and of opsonic capacity. Test group Plaque Index scores were higher than the control group on Days 7, 14, and 21, while the Gingival Index of the test group was elevated only on Day 21. Phagocytosis of test subjects' PMNs and opsonic capacity of their sera were no different from that of the controls'. Levels of C3 in both nonactivated and activated sera of the test group were significantly lower than that of the control group only on Day 14. In the test group, differences of per cent C3 conversion occurred between Days 14 and 21, and Plaque Index scores were inversely correlated with the order of per cent C3 data on these same days. Our results also indicate that complement contributes significantly to the serum opsonic capacity. The levels of developing plaque and subsequent gingival inflammation are apparently related to the degree of complement activation.

Adult

The quality of blood used for transfusion.

The current risk of disease transmission through homologous blood transfusion has lead to a revival in the use of autologous blood. The development of a coagulopathy increases the usage of blood and blood products and therefore the risk of disease transmission. Blood salvaged at operation is subjected to physical and humoral activating factors. The potential systems to be activated are located either in the plasma or non red cell cellular elements. Homologous blood and blood salvaged at operation before and after washing was examined for activation of the plasma and cellular systems by measuring the presence of fibrin degradation products (xDP's), activated complement (C3a) and plasminogen activator and inhibitor activity from the plasma systems; and elastase, serotonin, lysophospholipids, leukotriene B4, lysoplatelet factor and phospholipase A2 from the cellular systems. The plasma systems were activated in the salvaged blood which had elevated levels of xDP's and C3a compared to the patient's blood (p less than 0.05). The products of cellular activation were also elevated in operatively salvaged blood (p less than 0.01 for all the above products). The levels xDP's were normal but the levels of C3a increased in stored blood. The levels of the cellular systems, elastase, serotonin and lysoplatelet activating factor, increased with the duration of storage of bank blood. Unwashed and aged stored blood contains potentially harmful products. Consideration should be given to removal of non red cell elements in blood stored for surgery.

Blood Cells

Effect of anticoagulant on biocompatibility in membrane plasmapheresis.

The effect of heparin or citrate anticoagulation on blood cellular, complement pathway and coagulation pathway was investigated in a membrane plasma exchange procedure. Two membrane plasma separators constructed of cellulose di-acetate (CA) and polyvinyl chloride (PVC) were evaluated with heparin or citrate alone for anticoagulation in a 26 year old male with myasthenia gravis. Maximum white blood cell counts decrease was 21% at 30 min for the CA with heparin while no decrease was observed for the other schemes. Platelet counts changes were comparable between heparin and citrate, while the PVC groups showed less changes than the CA groups. The CA groups, regardless of the type of anticoagulant used, indicated that complement activation occurred via the classical pathway within the module in addition to the activation via alternative pathway for the CA with heparin. In the PVC groups, complement activation was noted to occur only when heparin was used for anticoagulation. PT and PTT showed slight increases with citrate, while they were remarkably prolonged with heparin. Citrate showed less changes in cellular and humoral factors compared to heparin. CA with heparin was the most activating combination of membrane material and anticoagulant, while the PVC with citrate was the least activating combination.

Adult

[Allergic reaction and vascular permeability].

The allergic reaction is characterized by an inflammatory phenomenon that comprises a cellular aspect and a vascular aspect. The vascular aspect is linked to the production of several mediators able to induce a vascular hyperpermeability. Among these mediators it is possible to find histamine, prostaglandins, leukotrienes, PAF, neuromediators, kinins, anaphylatoxins.

Animals

Structure and function of human C5a anaphylatoxin. Selective modification of tyrosine 23 alters biological activity but not antigenicity.

Reaction of either human C5a or its des-Arg74 derivative (des-Arg74-C5a) with tetranitromethane under nondenaturing conditions results in selective nitration of only 1 of the 2 tyrosine residues found in these glycopolypeptides. This reactive tyrosyl residue was identified as that found in position 23 of the sequence. Nitrotyrosyl23-C5a and -des-Arg74-C5a were separated from their respective unmodified precursors by cation-exchange fast protein liquid chromatography. These purified derivatives served as reagents for the subsequent preparation of both aminotyrosyl23-C5a and -des-Arg74-C5a as well as photoreactive analogs of C5a. Radioimmunoassays performed with C5a derivatives serving as competing ligands and a murine antihuman C5a monoclonal antibody employed as first antibody demonstrated that selective modification of tyrosine23 did not produce a detectible alteration in the antigenic properties of C5a. By contrast, either nitro- or aminotyrosyl23-C5a was approximately 3-fold less active than native C5a in both bioassays and competitive ligand-receptor binding assays. Additionally, photoreactive derivatives prepared by coupling either N-succinimidyl-6-(4'-azido-2'-nitrophenylamino)-hexanoate or p-nitrophenyl-2-diazo-3,3,3-trifluoropropionate to aminotyrosyl23-C5a at pH 5.0 failed to interact with the neutrophil C5a receptor. These observations suggest that the tyrosyl23 residue of C5a may participate importantly in the binding interactions of this chemotactic factor and its granulocyte receptor.

Affinity Labels

Human erythrocytes inhibit complement-mediated solubilization of immune complexes.

Incubation of precipitable immune complexes (IC) with fresh human serum or guinea pig serum resulted in solubilization of IC. When packed human E were added to human serum or guinea pig serum, binding of IC to the E occurred and IC solubilization was significantly inhibited. By contrast, SRBC did not bind IC nor inhibit IC solubilization. Because IC binding to human E is mediated by CR type 1 (CR1) we evaluated whether CR1 was responsible for the inhibition of IC solubilization. Human E were treated with trypsin or anti-CR1 mAb. Both treatments abrogated IC binding to human E but did not affect the ability of the human E to inhibit IC solubilization. Human E inhibited C activation by IC. Thus, incubation of IC in human serum caused significant activation of C3 and C5, but not C4. However, when IC were incubated in whole blood or with isolated human E and serum, C3 activation by IC was inhibited significantly. In addition, we demonstrated that the C3b generated during C activation by IC deposited on both IC and human E. Thus, human E may compete for nascent C3 generated during C activation by IC. In conclusion, human E inhibit both complement-mediated solubilization of IC and C activation by IC.

Antigen-Antibody Complex

[The reactivity of pulmonary and hepatic macrophages in normal and prestimulated animals].

The rate of blood clearing of colloidal carbon increased two-fold as compared to the controls 5 days after prodigiosin injection. This was attended by intensified recruitment of macrophages into the alveoli and increase in the number of phagocytizing macrophages in the interstitium of the lungs by 1.5 times above normal. After administration of zymosan granules, the rate of colloid carbon clearance increased 1.5-fold in 5 days and 2.5-fold on the 9th day; the recruitment of macrophages into the alveoli and the number of phagocytizing macrophages in the interstitium of the lungs increased two-fold. At the same time, granuloma-type mononuclear infiltrates formed in the lungs and liver. Prodigiosin injection increases the rate of blood clearance 1.2-fold in animals stimulated by zymosan granules, while the volume of mononuclear infiltration, which acquires a diffuse character, increases 2.5-fold as compared to the value in zymosan stimulated animals. The effects of phagocyte reconciliation may be directly related to exacerbations of chronic inflammatory diseases of different localization.

Animals