Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Complement Pathway, Classical”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,639 records · Page 91Linked to original sources
PubMed2003

Anti-complement activity in the saliva of phlebotomine sand flies and other haematophagous insects.

The saliva of haematophagous insects has a series of pharmacological activities which may favour blood feeding. In the present study, an inhibitory effect on the complement system was observed in salivary extracts obtained from the phlebotomine sand flies Lutzomyia longipalpis and Lu. migonei. Saliva from Lu. longipalpis was capable of inhibiting both the classical and alternative pathways, while that from Lu. migonei acted only on the former. Other haematophagous insect species were screened for inhibition of the classical pathway. The triatomine bugs Panstrongylus megistus, Triatoma brasiliensis and Rhodnius prolixus were also able to inhibit the classical pathway whereas the mosquito Aedes aegyti and flea Ctenocephalides felis were not. The activity of Lu. longipalpis saliva on the classical pathway was partially characterized. The inhibitor is a protein of Mr 10000-30000 Da, which is very resistant to denaturation by heat. The inhibition of the complement system by phlebotomine sand flies may have a role in the transmission of Leishmania to the vertebrate hosts. The inhibitor molecule is thus a promising component of a vaccine to target salivary immunomodulators.

Animals↗
PubMed1994

Effect of Melia azedarach L. leaf extracts on human complement and polymorphonuclear leukocytes.

The effect of the water extract of Melia azedarach L. (Meliaceae) leaves on human complement and polymorphonuclear leukocytes was investigated. This extract showed a strong anticomplementary activity, which was more pronounced in the classical pathway assay. The extract did not affect the phagocytic activity of polymorphonuclear leukocytes, nor the respiratory burst of these cells as measured by the nitro blue tetrazolium reduction assay.

Cells, Cultured↗
PubMed1981

Selective deficiency of the fourth component of complement in a patient with systemic lupus erythematosus (SLE): immunochemical and biological studies.

A 45-year-old female (J.B.) with clinically inactive systemic lupus erythematosus was devoid of haemolytic serum complement activity. Functional and immunoprecipitin assays showed that the complement defect was due to lack of the fourth component of complement (C4). The patient's serum and red cells were Chido-negative, Rodgers-negative. In J.B.'s serum there was marked impairment of opsonization of Staphylococcus aureus and of the formation of chemotactic activity in the presence of zymosan. The chemotactic defect was not due to the presence of inhibitory factors: the activity was increased by the addition of C4. These findings suggest that an intact classical pathway is necessary for the optimal generation of serum chemotactic factors by zymosan via the alternative pathway of the complement system.

Adolescent↗
PubMed2005

Complement inhibition rescued mice allowing observation of transgene expression following intraportal delivery of baculovirus in mice.

BACKGROUND: The baculovirus Autographa californica nucleo-polyhedrosis virus (AcNPV) is an alternative to other viral vectors for hepatic gene delivery. A barrier to AcNPV being used in vivo is its susceptibility to inactivation by serum complement 1. In vivo utility has only been demonstrated using methods that avoid contact with serum 2-5. We have studied the complement pathways involved in baculovirus inactivation in vitro and the systemic administration of baculovirus vectors in vivo, with the co-administration of the complement inhibitor, soluble complement inhibitor 1 (sCR1). RESULTS: EDTA increased baculovirus survival in human serum more than EGTA showing that both the alternative and classical pathways of complement are activated. Depleting serum of IgM increased survival, whereas reconstitution with pooled IgM restored activity against baculovirus, suggesting naturally occurring IgM antibodies with affinity for baculovirus may be partially responsible for complement activation. Intraportal administration of baculovirus led to hepatic expression when the complement inhibitor sCR1 (soluble complement receptor type 1) was co-administered by tail vein injection; however, liver histology showed hepatic necrosis. Without co-administration of sCR1, intraportal infusion of baculovirus was fatal within 24 h. Histology demonstrated massive hepatic necrosis. Yolk sac vein injection of baculovirus was associated with fetal death. CONCLUSIONS: Transgene expression was demonstrated following intraportal infusion of recombinant baculovirus vectors in combination with sCR1; however, our experiments suggest a significant associated toxicity.

Animals↗
PubMed2006

Which complement assays and typings are necessary for the diagnosis of complement deficiency in patients with lupus erythematosus? A study of 25 patients.

INTRODUCTION: Deficiencies in components of the classical pathway of complement activation are strong risk factors for lupus erythematosus (LE).Yet, it has not been addressed whether the conventional measurements of the serum hemolytic CH50 activity and antigenic concentrations of C3 and C4 are sufficient to asses a deficiency in C4A, C4B or C2 components, the most common deficiencies associated with LE. PATIENTS AND METHODS: In a retrospective series, we performed complement analyses in 35 patients with LE who were systematically screened for a complement deficiency. The majority of patients had cutaneous LE with mild systemic involvement and no complement consumption. Of 25 patients (72%) with complement deficiency we found 13 with a partial C4A deficiency, 2 with a complete C4A deficiency, 6 with a partial C4B deficiency, 2 with a complete C4B deficiency and 2 with a combined partial C2 and C4A deficiency. RESULTS: The total complement activity (CH50) was decreased in only one out of two patients with complete C4B deficiency. CH50 level was found to be low-normal (35-38 U/ml(-1)) in one patient with partial C4B deficiency, one patient with complete C4B deficiency and both patients with combined partial C4A and C2 deficiency. Total C4 levels were normal in 9 out of 13 the patients with a partial C4A deficiency and in 2 out of 6 patients with a complete C4B deficiency. The antigenic concentration of C3 was low in only 1 patients with a complete C4B deficiency and within the normal range in all the others patients. Overall, 50% of the patients had normal or elevated C3, C4, and CH50 levels. DISCUSSION: This study emphasizes that the usual measurements of CH50, C3 and C4 levels are not adequate to detect a C4 and/or C2 deficiency in patients with LE. In epidemiologic or investigative studies addressing the prevalence of complement deficiency, more elaborated diagnostic tests, such as C4 protein allotyping, C2 level measurement and genetic screening for type I C2 deficiency should also be performed.

Adult↗
PubMed1983

Visualization of human C4b-binding protein and its complexes with vitamin K-dependent protein S and complement protein C4b.

C4b-binding protein (C4bp) participates in the regulation of the C3 convertase of the classical pathway of complement. By binding to C4b, which is one of the structural subunits of this enzyme, C4bp accelerates the decay-dissociation of the enzyme and renders C4b susceptible to degradation by factor I (C3b inactivator). C4bp is a high molecular weight plasma protein (Mr = 570,000) composed of apparently identical subunits (Mr = 70,000) linked by disulfide bonds. In plasma and in purified form C4bp also forms a bimolecular complex (Kd = 0.9 X 10(-7) M) with protein S, a recently identified vitamin K-dependent plasma protein. The binding sites on C4bp for protein S and C4b are distinct and noncompetitive and protein S does not influence the function of C4bp as a regulator of the C3 convertase. C4bp, C4b, and protein S were visualized by electron microscopy by negative staining. C4bp was found to have an unusual spider-like structure. It is composed of seven thin (30 A), elongated (330 A), and flexible subunits that are linked to a small central body. Protein S exhibited two globular domains of equal size with a center-to-center distance of approximately equal to 50 A. Protein S was found to bind to the C4bp through only one of its domains by attaching to a short subunit that is distinct from the other seven subunits. C4b imaged as an irregular, relatively compact molecule. It was found to interact with the peripheral ends of the elongated subunits, suggesting seven C4b-binding sites per molecule of C4bp.

Carrier Proteins↗
PubMed2002

Growth phase influences complement resistance of Bordetella pertussis.

The BrkA protein of Bordetella pertussis inhibits killing by the antibody-dependent classical pathway of complement; however, susceptibility to complement can be highly variable. Log-phase bacteria grown in Stainer-Scholte (SS) broth plated on Bordet-Gengou (BG) agar were about 500 times more sensitive to killing by complement than stationary-phase SS-BG cultures. While always more susceptible to complement than the wild-type strain, a BrkA mutant displayed a similar growth phase variation in susceptibility to complement. Growth phase susceptibility to complement was also observed for a mutant constitutive for Bvg activation of BrkA, suggesting that modulation of virulence factor expression was not responsible for sensitivity to complement. Susceptibility was not due to differential antigenic expression, since serum adsorbed with complement-resistant, stationary-phase SS-BG cultures lacked bactericidal activity against B. pertussis harvested at all times during the growth cycle. These results suggest that log-phase susceptibility to complement is not due to variable expression of BrkA or antigenic differences and may be an inherent property of rapidly growing cultures. Implications for vaccine development are discussed.

Antigenic Variation↗
PubMed2001

A novel interaction between type IV pili of Neisseria gonorrhoeae and the human complement regulator C4B-binding protein.

C4b-binding protein (C4BP) is an important plasma inhibitor of the classical pathway of complement activation. Several bacterial pathogens bind C4BP, which may contribute to their virulence. In the present report we demonstrate that isolated type IV pili from Neisseria gonorrhoeae bind human C4BP in a dose-dependent and saturable manner. C4BP consists of seven identical alpha-chains and one beta-chain linked together with disulfide bridges. We found that pili bind to the alpha-chain of C4BP, which is composed of eight homologous complement control protein (CCP) domains. From the results of an inhibition assay with C4b and a competition assay in which we tested mutants of C4BP lacking individual CCPs, we concluded that the binding area for pili is localized to CCP1 and CCP2 of the alpha-chain. The binding between pili and C4BP was abolished at 0.25 M NaCl, implying that it is based mostly on ionic interactions, similarly to what have been observed for C4b-C4BP binding. Furthermore, the N-terminal part of PilC, a structural component of pili, appeared to be responsible for binding of C4BP. Membrane cofactor protein, previously shown to be a receptor for pathogenic N. gonorrhoeae on the surface of epithelial cells, competed with C4BP for binding to pili only at high concentrations, suggesting that different parts of pili are involved in these two interactions. Accordingly, high concentrations of C4BP were required to inhibit binding of N. gonorrhoeae to Chang conjunctiva cells, and no inhibition of binding was observed with cervical epithelial cells.

Antigens, CD↗
PubMed2000

A cluster of positively charged amino acids in the alpha-chain of C4b-binding protein (C4BP) is pivotal for the regulation of the complement system and the interaction with bacteria.

C4b-binding protein (C4BP) is a potent inhibitor of the classical pathway of complement. Structural analysis of our 3D model of the C4BP alpha-chain suggested that a cluster of positively charged amino acids at the interface between complement control protein (CCP) modules 1 and 2 could be involved in ligand binding. Nine C4BP mutants, where the positively charged amino acids were changed to glutamines, were expressed. We found that all of these displayed lower apparent affinity for C4b and that the site was also a specific heparin-binding site. The mutants demonstrated decreased ability to serve as factor I cofactors in a degradation of C4b. Also, their capacity to prevent the assembly of C3-convertase and to accelerate its decay were decreased. In conclusion, we found a binding site on C4BP that is functionally important for the regulation of C3-convertase. Many strains of Streptococcus pyogenes bind C4BP, via surface M proteins, which plays an important role in pathogenesis. Using the set of C4BP mutants we identified a key recognition surface for M proteins which overlaps with the C4b-binding site. The analysis of all mutants and inhibition with monoclonal antibodies indicated that the binding sites for C4b and M proteins are only overlapping but not identical. Furthermore, we found that the molecular mechanisms involved in these two interactions differ, since the binding between M proteins and C4BP is relatively insensitive to salt in contrast to the C4BP-C4b-binding. We suggest that binding between C4b and C4BP is governed mostly by electrostatic interactions, while additional non-covalent forces cause tight binding of C4BP to streptococcal M proteins.

Amino Acids↗
PubMed1994

Inhibition of complement activation by natural sulfated polysaccharides (fucans) from brown seaweed.

In the present study, we demonstrate that natural sulfated polysaccharides (fucans) isolated from brown seaweed are potent inhibitors of human complement activation. A fucan fraction of chromatographic molecular weight 22,600, termed BS8, was found to inhibit classical and alternative pathway activation in whole serum in a dose-dependent fashion. Fucan BS8 inhibited formation of the classical pathway C3 convertase by interfering with C1 activation or by inhibiting C4 cleavage and the interaction between C4b and C2. The fucan also inhibited formation/function of the alternative pathway C3 convertase by suppressing the binding of B to C3b and by interfering with the stabilizing function of Properdin. The inhibitory effect of fucans on formation of the C3 convertases was dependent on the molecular weight of the polysaccharide for compounds of chromatographic molecular weight below 16,600. Fucan had no effect on the function of the terminal complex. Since fucans were more efficient than heparin in inhibiting activation of the classical pathway in whole serum and exhibited a lesser specific anticoagulant activity on a molar basis, our results suggest that these natural sulfated polysaccharides have a potential for use as anti-complementary and anti-inflammatory agents.

Complement Activation↗
PubMed1994

Development of a C1q-ABO-ELISA to measure C1q binding by human anti-A alloantibodies.

Little is known about the correlation between the amount of anti-A antibodies and the respective complement activating capacity. We have therefore developed a new ELISA procedure for the study of complement activation by anti-A antibodies bound to immobilized blood group substance A through the classical pathway. C1q binding capacity of anti-A was then compared to both anti-A immunoglobulin (Ig) isotype serum concentrations and to the hemolysing capacity of O type sera. We have demonstrated that sera with low anti-A IgG and IgM levels produced low C1q binding activity and no hemolysis of human A1 red blood cells (RBC), whereas higher anti-A Ig levels resulted in either weak or strong C1q binding capacity as well as weak or strong RBC hemolysis. A correlation of rs = 0.755 was observed between C1q-ABO-ELISA and hemolysin assay results, although only 58% of the O type sera containing C1q binding anti-A lysed A1 type RBC under the used conditions. The C1q-ABO-ELISA is therefore a more sensitive assay for detecting 'dangerous' O type donors for blood transfusion to A type recipients than screening tests for hemolytic anti-A or determination of anti-A Ig levels alone.

ABO Blood-Group System↗
PubMed1984

Serial determination of complement and specific antibody titres in Pseudomonas aeruginosa infection.

Serial examination of the complement system and specific antibody titre of 12 surviving patients and 8 lethal cases suffering from Pseudomonas aeruginosa infection showed that except the onset of infection, until the development of septic shock, the level of the complement components corresponded to or exceeded the average normal value. In reversible septic shock the complement titre decreased significantly and in irreversible shock the values were even lower. Activation of the complement system occurred on 10 occasions via the classical and on 42 occasions via the alternative pathway. The number of activations grew parallel with the severity of the infection. Activation through the classical way was generally more intensive. During the whole infectious process not the individual characteristics of the P. aeruginosa present unbroken, but the pathological events and the specific antibody level determined the mode (alternative or classical pathway) of complement activation. The specific antibody level of the surviving patients significantly surpassed the titres of the lethal cases until the development of shock. Not an insufficiency of the complement system but the relative lack of specific antibodies was mainly responsible for the fatal outcome of P. aeruginosa infections.

Antibodies, Bacterial↗
PubMed2005

A complement receptor for opsonized immune complexes on erythrocytes from Oncorhynchus mykiss but not Ictalarus punctatus.

Immune complexes activate the classical pathway of complement resulting in the covalent deposition of fragments of the third (C3b) and fourth (C4b) components of complement, thus opsonizing the complexes for uptake by CD35 found on human erythrocytes. The complexes are then transported to and cleared from the circulation by the reticuloendothelial system. It has been shown that rainbow trout can remove immune complexes from the circulation in a complement-dependent manner similar to that found in the human. However, the cell or cell types involved have not been identified. The purpose of this study was to investigate whether a complement-dependent immune adherence receptor is expressed on erythrocytes from the rainbow trout (Oncorhynchus mykiss) and the channel catfish (Ictalarus punctatus). Coating fluorescent microparticles with BSA, and then binding them to anti-BSA created an artificial immune complex that was incubated with normal fish serum, normal human serum or EDTA-treated serum. The complement-coated immune complexes were then incubated with either fish or human erythrocytes and analyzed for binding by flow cytometry and further visualized by fluorescence microscopy. Our results indicate that erythrocytes from rainbow trout are capable of binding immune complexes when pretreated with serum from either the trout or human, but not when pretreated with serum containing EDTA. By contrast, erythrocytes from the channel catfish did not bind immune complexes pretreated with autologous or human serum. These data suggest that differences exist in receptor distribution between two closely related species of fish, and a potentially homologous relationship in receptor expression, and possibility function, exist between two highly divergent species.

Animals↗
PubMed1980

A simple immunofluorescence assay for C1q binding.

Antibody-independent binding of B1q to Crithidia luciliae kinetoplast DNA was demonstrated by immunofluorescence microscopy. The method allows detection of C1q in human sera diluted 1/20--1/40 and in 5--10 micrograms/ml concentrations of isolated C1q. Various substances known to interfere with the early events of complement activation inhibited binding of C1q to kinetoplasts. The method may be used as a functional assay for serum C1q.

Binding Sites, Antibody↗
PubMed1999

C1q binding to liposomes is surface charge dependent and is inhibited by peptides consisting of residues 14-26 of the human C1qA chain in a sequence independent manner.

Complement activation by anionic liposomes proceeds by antibody-independent, C1q-initiated activation of the classical pathway. Purified C1q bound to anionic liposomes in an acidic lipid concentration-dependent manner. Saturation binding, but not the apparent association constant, was enhanced by increasing the cardiolipin content of the liposomes or decreasing either the pH or ionic strength of the reaction mixture. These observations indicate the involvement of electrostatic factors in the binding. A highly cationic region in the collagen-like domain of C1q comprised of residues 14-26 of the C1qA polypeptide chain was assessed for involvement in liposome binding. This region has previously been shown to mediate C1q binding to other immunoglobulin-independent activators of the classical pathway of complement. Peptides containing residues 14-26 of C1qA, denoted C1qA14-26, inhibited C1q binding to and complement activation by anionic liposomes. The inhibitory capacity of these cationic peptides had no sequence or conformation specificity. Rather, the amount of positive charge on the peptides was the determining factor. When present in excess, peptides with five cationic residues inhibited C1q binding and complement activation; however, C1q peptides with only two cationic residues did not. In addition to the C1qA14-26 region, other parts of C1q that contain cationic residues may also be involved in C1q binding to anionic liposomes.

Complement Activation↗
PubMed2002

Structural biology of the C1 complex of complement unveils the mechanisms of its activation and proteolytic activity.

C1 is the multimolecular protease that triggers activation of the classical pathway of complement, a major element of antimicrobial host defense also involved in immune tolerance and various pathologies. This 790,000 Da complex is formed from the association of a recognition protein, C1q, and a catalytic subunit, the Ca2+-dependent tetramer C1s-C1r-C1r-C1s comprising two copies of each of the modular proteases C1r and C1s. Early studies mainly based on biochemical analysis and electron microscopy of C1 and its isolated components have allowed for characterization of their domain structure and led to a low-resolution model of the C1 complex in which the elongated C1s-C1r-C1r-C1s tetramer folds into a more compact, "8-shaped" conformation upon interaction with C1q. A major strategy used over the past years has been to dissect the C1 proteins into modular segments to characterize their function and solve their structure by either X-ray crystallography or nuclear magnetic resonance spectroscopy (NMR). The purpose of this review is to focus on this information, with particular emphasis on the architecture of the C1 complex and the mechanisms underlying its activation and proteolytic activity.

Animals↗
PubMed1989

Interaction of campylobacter species with antibody, complement and phagocytes.

The opsonisation of four different campylobacter species for human neutrophils was studied using a chemiluminescence system and electron microscopy. Opsonisation of Campylobacter fetus, Campylobacter coli, and Campylobacter jejuni was mediated by antibody and enhanced by complement. Antibody was not, however, required for the phagocytosis of Campylobacter pylori because it activates the classical pathway of complement directly. This unusual property may be important in the pathogenesis of C pylori associated gastritis and duodenal ulcer.

Anaphylatoxins↗