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Chemotactic factor generation by asbestos. Fibre type differences and the effects of leaching.

The generation of chemotactic factors in normal human serum by treatment with chrysotile and crocidolite asbestos was investigated using the Boyden chamber method. Both types of asbestos produced chemotactic activity but not to the same extent. The two fibre types also produced different levels of complement activation as measured by C3 conversion using two dimensional immunoelectrophoresis. While there was a simple correlation between fibre concentration and C3 conversion the production of chemotactic activity declined at high doses of chrysotile. Phosphate buffer treatment reduced the differences between fibre types. The possible role of these phenomena in asbestos-associated disease is discussed.

Asbestos↗

[Facial eosinophilic granuloma].

We report on a patient with the diagnosis granuloma eosinophilicum faciale (GEF). Clinical and histological findings are described with special emphasis on immunhistological characteristics in lesional skin. A review on the literature is given.

Adult↗

Autoimmune phenomena in infertile patients with endometriosis.

To assess the presence of autoimmune factors in patients with endometriosis, endometrial homogenates, peritoneal fluid, and serum were evaluated in 23 patients with endometriosis and 22 control subjects. The methods used were Ouchterlony immunodiffusion, immunoelectrophoresis, polyacrylamide gel electrophoresis, and radial immunodiffusion. The study demonstrated precipitation lines between endometrial homogenates and serum from some endometriosis patients but not from the control subjects. Immunoelectrophoresis demonstrated precipitation lines at beta-globulin position when endometrial homogenates were used against serum of patients with endometriosis and with goat antihuman serum. It is suggested that an antigen, possibly a glycoprotein, is present in the endometrial homogenates. Radial immunodiffusion studies for immunoglobulins (Ig) G, A, and M and complement components C3c and C4 showed significantly higher concentrations of C3c and C4 in serum and peritoneal fluid of patients with endometriosis than the control subjects (P less than .05). There was no significant difference in concentration of IgG, A, M, factor B, and properdin.

Adolescent↗

Association of rheumatoid factor with complement activation in rheumatoid arthritis and other diseases.

Rheumatoid factor (RF) is a complement activating autoantibody. In rheumatoid arthritis (RA) the rate of catabolism of complement is closely related to the titre of RF. Therefore, we have examined whether these relationships are unique to RA or will be found in non-RA disorders in which RF may be found in the circulation. We studied patients with subacute bacterial endocarditis, leprosy, tuberculosis, and a variety of other rheumatic and vasculitic disorders. We found that in all the disorders examined the RF had a complement activating potential which was equivalent to that of the RF of RA patients. Furthermore in vivo activation of complement, as exhibited by the appearance of C3 degradation products, was significantly related to higher titres of haemolytically active RF in non-RA as well as the RA group. In these respects, therefore, the RF in RA and non-RA patients is indistinguishable. A possible survival value for RF is discussed.

Adolescent↗

Humoral and cellular immunity in extrinsic pulmonary granulomatoses.

In the present study the AA. analyse the immunological behaviour of patients taken from a group of 309 cases of Extrinsic Pulmonary Granulomatoses (E.P.G.). Humoral immunological changes were studied by assessing serum immunoglobulins, various complement fractions and, when possible, circulating precipitating antibodies. Immunoglobulins A and G values were significantly higher than control values. On the contrary there was no significant difference between IgM values, in patients and controls. The complement fractions (C3c, C4 and C3 activator) showed values within normal limits. Specific circulating antibodies were detected both in bird fanciers and in cork workers. Cellular immunity was evaluated by comparing the results of various tests. Eighty percent of all cases of E.P.G. had negative tuberculin tests, while eighty percent of the general population, in Portugal, has positive tuberculin tests. The Kveim-Siltzbach test was positive in one of 19 cases of E.P.G., and this was considered a false positive result. In vitro lymphocyte blastic transformation in presence of pigeon antigen showed a significant increase in pigeon breeders as compared with controls. The migration of leucocytes from affected and non-affected corkworkers, affected and non-affected pigeon fanciers and controls, in the presence of the specific antigens, showed significant results. Finally, similarities and differences in immunological behaviour of these patients are discussed.

Antibody Formation↗

[Mechanisms and implications of the adhesion phenomenon in Aspergillus fumigatus].

During the last few years, several works have demonstrated the fixation of different host proteins on Aspergillus fumigatus conidia. Thus, after incubation in the presence of normal human plasma, the C3 component of complement is detected at the surface of conidia. In fact, most of the C3 deposited on conidia is converted in C3b or iC3b which would facilitate their phagocytosis by the macrophages. In the non immune host, the activation of the alternative pathway seems to be the main mechanism of the activation of the complement system by the conidia, but the participation of the classical pathway initiated by the fixation of the C-reactive protein has also been suggested. Aspergillus fumigatus conidia interact also with fibrinogen and laminin. These interactions which are mediated by the D domains of fibrinogen and by the fragment P1 of laminin, are specific. The number of fibrinogen binding sites at the surface of conidia has been calculated to be 1200 by cell, and the dissociation constant 2.2 x 10(-7) M. These interactions could determine the adhesion of conidia to the host tissues as suggested by adherence assays of conidia to proteins immobilized onto wells of microtiter plates. Conidia would bind to the fibrin deposits formed on damaged epithelia in response to the inflammatory reaction, or directly to laminin of the subepithelial basement membrane. Finally, different experiments suggested the identity of the binding sites for C3, fibrinogen and laminin at the surface of A. fumigatus conidia.

Aspergillus fumigatus↗

The time courses of intracellular transport of some secretory proteins of rat liver are not affected by an induced acute phase response.

To study the effects of changed synthesis on intracellular transport of secretory proteins, an acute phase response was induced in rats. The synthesis and secretion of haptoglobin, complement C3, transferrin and albumin were then investigated by pulse labeling with [3H]leucine. Maximal increase in the syntheses of the positive acute phase proteins was observed after 24 h, amounting to an increase of nine, three and twofold for haptoglobin, C3 and transferrin, respectively. The synthesis of albumin decreased to a minimum after 48 h, reaching approximately one fourth of normal synthesis. The time courses for transit through rough endoplasmic reticulum and for secretion were determined after 36 h, and were found to be roughly unchanged for all four proteins despite the different changes in synthesis. The fraction of haptoglobin associated with the microsomal membrane was reduced during the acute phase response, but there was no significant change in membrane association as a function of time after labeling with [3H]leucine. It is concluded that the altered protein synthesis during an acute phase response in vivo has little effect on the time course of secretion of the proteins studied. Furthermore, the basal mechanisms for intracellular transport appear relatively unchanged during this condition.

Acute-Phase Reaction↗

Some immunological aspects on human Schistosoma mansoni infection in Egypt.

Human Schistosomiasis mansoni is a chronic disease, which ranges from subclinical infection to huge hepatosplenomegaly. Reducing the risk of infection remains the aim of many control measures. So the present study was designed to evaluate the interaction between S. mansoni infection and immunological system of the host. The study included 3 groups of 50 adult males each. The first group was control group, the second group was early S. mansoni infection and third group was advanced schistosomiasis with hepatosplenomegaly. The results showed some changes in both humoral and cellular immune response during the course of infection.

Adolescent↗

[The effects of occupational exposure to nitrogen dioxide (NO2) on the immunological parameters in workers].

The study of T-cell subpopulation: T CD3+, T CD4+, T CD8+; NK CD16+ and B CD19+ cells as well as of serum concentrations of immunoglobulins G, A, M, E, complement components C3c and C4, total circulating immune complexes (CIC), CRP, haptoglobin, ceruloplasmin and transferin, was carried out in 16 males, smokers and non-smokers. The control group of smokers and non-smokers comprised 36 males not exposed to any chemical compound. The study was performed also in two groups of non-smokers: 5 workers occupationally exposed and 7 non-exposed controls. For the determination of T-cells, NK- and B CD19+ cells populations monoclonal antibodies were used in indirect immunofluorescence tests. Passive sampling spectrophotometric method was employed to determine NO2 and NO concentrations in the work environment air. Their mean values were 0.0867 +/- 0,0585 mg x m-3 (from 0.0165 to 0.1960) and 0.0614 +/- 0.0263 mg x m-3 (from 0.0220 to 0.1090), respectively. The number of total leucocytes and lymphocytes in the group of 16 smokers and non-smokers exposed to NO2 and NO was increased. Stimulation T-cells line in the men exposed to NO2 and NO was evidenced by an increased number of T CD3+ cells, about two-fold increase in absolute number of T CD4+ cells (p < 0.001), an increased number of T CD8+ cells (p < 0.001) and by an enhanced value (by 24.7%) of the T CD4+/T CD8+ ratio. In the group of 16 persons exposed to NO2 and NO serum IgG concentration was elevated (p < 0.01), C3c C4 decreased (p < 0.001), and serum CIC enhanced by about two times (p < 0.001). Stimulation T-cells line in the groups of 5 non-smoking workers exposed to NO2 and NO was evidenced by an increased number of T CD3+ (p < 0.05), T CD4+ (p < 0.05) and T CD8+ cells but without any change in the value of the T CD4+/T CD8+ ratio. In addition, decreased serum C3c and C4 levels together with serum CIC elevated by 76.7% (p < 0.05) were evidenced in this group on non-smokers. Moreover, significant positive correlations between NO2 concentrations in the air and the numbers of total lymphocytes, T CD3+, TCD4+, T CD8+ cells or IgG: (magnitude of r in the range between 0.31 and 0.71), as well as significant negative correlations between NO2 concentrations in the air and C3c (r = 0.44) in the group of 16 smoking and non-smoking workers were calculated. In this group significant positive correlation between NO concentrations in the air of the workplace and counts of T CD3+, T CD8+, B CD19+ (magnitude of r in the range between 0.51 and 0.63), haptoglobin and ceruloplasmin (magnitude of r from 0.37 to 0.58), as well as significant negative correlation between NO concentrations in the air and serum levels of IgG, IgA and IgM (magnitude of r from -0.67 to -0.47) and CIC (magnitude of r = -0.39) were also observed. In the group of 5 exposed non-smoking workers, the values of correlation coefficients magnitude of r between NO2 and NO concentrations in the work environment air and immunological parameters were similar to those in the whole group. The results obtained suggest that during occupational exposure NO2 may play a more important role mainly in the process of inflammation but exogenous NO seems to act as modulating factor of this proinflammatory NO2 effect through a greater and exposure-dependent influence mainly on B CD19+ cells and other parameters of humoral immunity.

Adult↗

Activation and fragmentation of the third (C3) and the fourth (C4) components of complement: generation and isolation of physiologically relevant fragments C3c and C4c.

The degree of the activation and fragmentation of C4 and C3, including chain structure of the activation products, was evaluated by SDS-PAGE analysis of the C4 or C3 antigens that were withdrawn from the reaction media with appropriate immunoadsorbent beads. Full activation of C4 and C3, and subsequent quantitative conversion of C4b into C4c, and C3b into iC3b took place in fresh NHS after the activation of complement with both aggIgG and CVF. For complete conversion of iC3b to C3c erythrocytes carrying the C3b receptor were added to the already activated serum. Both C4c and C3c were isolated by a 2-step procedure involving (i) an adsorption to and (ii) electrophoretic desorption from the respective immunoadsorbent beads.

Complement Activation↗

Temporal sequence of changes in tear film composition during sleep.

Overnight eye closure induces a shift in the nature and composition of the tear film, from a dynamic reflex tear-rich to a stagnant secretory IgA-rich layer. This is accompanied by the induction of a state of sub-clinical inflammation, as evidenced by increases in albumin levels, plasminogen activation, conversion of complement C3 to C3c, and the recruitment of polymorphonuclear (PMN) cells into the tear film. To determine the time course and functional relationship between these potentially interdependent processes, tear samples were collected from ten non-contact lens wearers after 1, 2, 3 and 5 hours of sleep. A subgroup of 6 subjects also self-collected tear samples after 8 hours of sleep. Tear samples were analysed for albumin by quantitative immunofixation assay, secretory IgA (sIgA) by radial immunodiffusion assay, plasmin-like activity using a chromogenic substrate, and complement C3 to C3c conversion by immunoblot assay. Epithelial and PMN cells in the precorneal tear film were recovered from corneal washings from the same subjects after 1, 3, 5 and 8 hours of sleep, and quantified. Results revealed that, unlike epithelial cells which exhibited a slow progressive accumulation as a function of the period of sleep, PMN cell concentration exhibited a lag phase, with recruitment occurring after between 3 and 5 hours of eye closure. This was preceded by plasminogen activation, increases in albumin and sIgA levels, and complement C3 to C3c conversion, all of which occurred within 1 to 3 hours after eye closure. Plasmin-like activity appeared to plateau after 3 hours and then decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Analysis of corneal inflammation following the injection of heterologous serum into the rat cornea.

The immunopathologic response following the injection of various antigens into the rat cornea was evaluated. This reaction, known as Wessely's phenomenon, was believed to be primarily triggered by antibodies and complement activation. The keratitis model was originally described in rabbits, using heterologous serum or purified proteins. In rats only, heterologous serum induced corneal inflammation with the characteristics of Wessely's phenomenon, (ie, a quiescent period of several days between antigen injection and onset of clinical signs and corneal opacification). Using rats allowed us to characterize the cellular infiltrate with immunohistochemical methods. Marked infiltration of the cornea by macrophages was observed, as was infiltration by polymorphonuclear cells, although to a lesser extent. Furthermore, T lymphocytes of the helper phenotype were demonstrated. Antibodies to complement activation product C3c showed faint staining, whereas B lymphocytes and plasma cells were absent. In addition, inflammatory cells and ocular tissues, particularly the limbal and peripheral corneal epithelium, were found to express major histocompatibility complex class II antigens during the inflammatory response. After the inflammation had subsided, macrophages and T lymphocytes remained in the corneal stroma (at least until day 30). These findings suggest that antigen-induced keratitis in rats might be mediated, at least partially, by T helper lymphocytes.

Animals↗

Diurnal tear cycle: evidence for a nocturnal inflammatory constitutive tear fluid.

To investigate the tear film in the closed eye, microliter tear samples were collected without overt reflex stimulation throughout the diurnal cycle, with closed eye samples recovered immediately upon eye opening. Samples were subjected to agarose, polyacrylamide, and two-dimensional electrophoresis, coupled with immunofixation, immunoblot, and lectin blot assays. Major protein constituents were densitometrically and immunologically quantified. Results revealed a distinct progression in composition from reflex to open to closed eye tear samples. Total protein increased from 6.0 to 9.0 to 18.0 mg/ml, secretory IgA increased from less than 0.23 to 0.85 to 8.40 mg/ml, and serum albumin increased from 0.02 to 0.06 to 1.10 mg/ml. In contrast, concentrations of the major reflex tear components (lysozyme, lactoferrin, and tear specific prealbumin) remained essentially static. Immunoblot assay for complement C3 and C3c revealed that eye closure was associated with C3 activation. Results indicate that: (1) the reflex and closed eye tear layers represent opposite extremes in composition and likely origins, with open eye tear film suggesting an intermediate origin; (2) reflex tears are derived from a neurologically inducible lacrimal or accessory gland secretion composed almost exclusively of lysozyme, lactoferrin, tear specific prealbumin, and a minor mixed alpha to beta globulin fraction; (3) upon eye closure, reflex secretion ceases or greatly diminishes, with ongoing slower flow maintained by a constitutive secretion composed almost exclusively of secretory IgA; (4) the closed eye environment induces a subclinical inflammation, accounting in part for the marked rise in albumin concentration. This increase, coupled with that of secretory IgA, may play a critical role in protecting the closed eye environment from pathogens. However, this may render the closed eye environment particularly vulnerable to inflammatory and immune-mediated pathological processes, such as those seen with extended wear soft contact lenses.

Adult↗

Glomerular C3c localization indicates ongoing immune deposit formation and complement activation in experimental glomerulonephritis.

In antibody-mediated glomerular disease, deposits of C3 (C3b) are common and are degraded by factor I to C3c and C3d. However, the kinetics of C3b degradation in glomerulonephritis have not been defined. To do this, we studied three models of complement-dependent glomerulonephritis with established C3 deposits (passive Heymann nephritis, cationized immunoglobulin G membranous nephropathy, and concanavalin A-anticoncanavalin A glomerulonephritis). C3b deposition was halted by administration of cobra venom factor, and the disappearance of C3c and C3d from glomeruli was measured with specific antibodies and quantitative fluorescence densitometry. Results showed that C3c deposits were reduced by over 85% within 24 hours in all three models. C3c clearance was unaffected by site or mechanism of deposit formation. C3d deposits persisted despite lack of ongoing complement activation. In passive Heymann nephritis when disease activity was monitored by urinary C5b-9 excretion, C3c was cleared in parallel with return of urine C5b-9 excretion to normal values. We conclude that glomerular deposits of C3c are cleared within 24 hours of cessation of complement activation. Positive staining for C3 utilizing antibody specific for the C3c portion documents recent complement activation usually reflecting new immune deposit formation.

Animals↗

Effect of antigen site and complement receptor status on the rate of cleavage of C3c antigen from red cell bound C3b.

C3b was bound to human red cells when serum complement was activated by addition of antibodies directed against different red cell antigens, and the rate of cleavage to C3dg was determined by assay for loss of bound C3c antigens using radiolabeled monoclonal anti-C3c. When C3b was bound by antibodies to antigens on branched-chain glycoproteins, cleavage to C3dg occurred more rapidly than when C3b was bound by antibodies to antigens closer to the red cell lipid bilayer. The rate of cleavage to C3dg also correlated directly with the number of complement receptors (CR1) per red cell, reflecting their role as cofactors in the cleavage of iC3b by factor I. Thus, the life span of C3b/iC3b on human red cells, which may be important for determining the rate and mechanism of clearance of C3-coated red cells, appears to depend on the CR1 status of the red cells and the characteristics of the antigen sites around which complement is bound.

Antigens↗

Acute complement activation during an anaphylactoid reaction to blood transfusion and the disappearance rate of C3c and C3d from the circulation.

A patient experienced an anaphylactoid shock after receiving a transfusion with 5-10 ml of leucocyte-depleted blood. Complement measurements revealed pronounced elevation of split products of the third complement factor (C3c and C3d), normal but fluctuating levels of native C3, significantly decreased levels of C4, normal factor B concentrations and low levels of factor B split products. The results indicate complement activation via the classical pathway as a causative mechanism of the anaphylactoid shock, and suggest an antigen-antibody reaction as the etiological factor. The nature of the antigen-antibody reaction initiating the complement activation could not be established. The disappearance rate (T/2) of C3d from the plasma after the acute C-activation was measured to approximately 4 hr, and the half life of C3c was significantly shorter.

Adult↗