RESUMO
OBJECTIVE: To investigate whether neutrophils and systemic complement are activated in pregnancies complicated by preeclampsia more than in normal pregnancies. METHODS: We measured native complement components and activation products in plasma by enzyme immunoassays in 19 women with uncomplicated pregnancies, 15 with preeclampsia before cesarean deliveries, and 16 nonpregnant women. Neutrophil activation was measured by specific enzyme immunoassays for myeloperoxidase and lactoferrin. RESULTS: Myeloperoxidase was significantly higher in women with preeclampsia (197 microg/L, 95% confidence interval [CI] 94, 646) than in women with uncomplicated pregnancies (124 microg/L, 95% CI 70, 289; P =.009), whereas lactoferrin did not differ between groups. C4 was decreased in preeclamptic women (0.16 g/L, 95% CI 0.07, 0.48) compared with women with uncomplicated pregnancies (0.21, 95% CI 0.10, 0.30, P <.001). There were no differences for the other native complement components. There was a significant decrease in C1rs-C1 inhibitor, 13 AU/mL (95% CI 9, 34) versus 19 (95% CI 13, 38) (P < or =.001) in normal pregnant women compared with nonpregnant women. There also was an increase in C3, C4, C9 (data not shown), C4bp, 132% (95% CI 94%, 161%) versus 91% (95% CI 57%, 128%); C3bc (7.4 AU/mL, 95% CI 4.2, 10.7) versus 4.8 AU/mL (95% CI 3.2, 7.3) and C4bc (8.6 AU/mL, 95% CI 5.7, 14.0) versus 3.5 AU/mL (95% CI 2.2, 6.7) in normal pregnant women compared with nonpregnant women (P < or =.001). CONCLUSION: Neutrophil activation in preeclampsia was shown by systemic increases in myeloperoxidase. Except for a decrease in C4, systemic complement activation could not be detected in preeclampsia.
Assuntos
Ativação do Complemento , Proteínas do Sistema Complemento/metabolismo , Neutrófilos/metabolismo , Pré-Eclâmpsia/sangue , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Técnicas Imunoenzimáticas , Lactoferrina/sangue , Peroxidase/sangue , GravidezRESUMO
BACKGROUND AND AIMS: Helicobacter pylori is a frequent gram-negative colonizer of the human stomach. Its interaction with complement may be involved in the pathogenesis of chronic gastritis, and was mechanistically studied in vitro. METHODS: Four H. pylori strains, 2 cytotoxin-associated genes (cag)A+ and 2 cagA-, were isolated from infected patients. Bacteria or purified H. pylori lipopolysaccharides (LPSs) were incubated with nonimmune serum at 37 degrees C; the activation products C3b/iC3b/C3c (C3bc) and terminal complement complex (TCC) were then quantified by immunoassays. The serum sensitivity of 1 strain (L01, cagA+) was tested by counting the numbers of colony-forming units. RESULTS: All strains and LPSs generated large amounts of C3bc and TCC. Blocking of the classic complement pathway by the calcium chelator ethylene glycol tetraacetic acid (EGTA) markedly reduced the complement products, suggesting that H. pylori and its LPSs directly engage the classic activation pathway. H. pylori was shown to be serum sensitive, but 30% or more nonimmune serum was necessary to induce marked killing. After 5 minutes, swelled bacteria coated with C3bc and TCC were shown. CONCLUSIONS: H. pylori is complement sensitive and activates the classic pathway even in the absence of specific antibodies. Released cell wall constituents such as LPSs can activate complement and may explain why this bacterium induces gastric pathology without invading the mucosa.
Assuntos
Antígenos de Bactérias , Ativação do Complemento/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/genética , Proteínas Sanguíneas/farmacologia , Ativação do Complemento/efeitos dos fármacos , Complemento C3b/imunologia , Complemento C3c/imunologia , DNA Bacteriano/análise , Imunofluorescência , Helicobacter pylori/genética , Humanos , Técnicas In Vitro , Lipopolissacarídeos/farmacologiaRESUMO
The interaction between a pectin type polysaccharide fraction, PMII, isolated from the leaves of Plantago major, and human complement was tested in two different hemolytic complement-fixation tests and in addition by two ELISA methods detecting complement-activation products. Sera were used as a complement source of 10 arbitrary human volunteers, individually and as a pool. The complement-fixation tests were designed to measure the concentration of the pectin necessary to inhibit 50% of the hemolysis (ICH(50)). The ELISA tests for complement-activation products were measured in AU/mg using a fully activated serum as a standard. We observed a more than 200-fold difference in ICH(50) activity of the PMII pectin in one of the hemolytic tests by varying the individual sera used as complement-source. On the other hand, the ELISA complement-activation tests showed no significant variation in activity of the PMII depending on the complement-serum used. The level of antibodies against PMII detected in the complement-sera did not correlate with the ICH(50) activity of PMII. The results show that PMII is a potent complement activator with an activity of the same order of magnitude on a weight basis as that of aggregated human immunoglobulin (Ig)G. This activation leads to a complement consumption probably explaining the PMII's effect in the complement-fixation tests. PMII seems to be an activator both on the classical and the alternative pathway of activation. The results might be related to the reported wound-healing effect of the leaves of Plantago major.
Assuntos
Ativação do Complemento , Pectinas/farmacologia , Folhas de Planta/química , Plantago/química , Plantas Medicinais , Adulto , Testes de Fixação de Complemento , Via Alternativa do Complemento , Via Clássica do Complemento , Relação Dose-Resposta a Droga , Feminino , Humanos , Magnoliopsida/química , Masculino , Medicina Tradicional , Pessoa de Meia-Idade , FitoterapiaRESUMO
Native complement factors and complement activation products were measured in healthy neonates (n = 72) and in a group of infants with premature prolonged rupture of the membranes (PPROM) without sepsis (n = 10). Vitronectin concentration in normal cord blood was not correlated with gestational age, and the median value was 86.0% of adult values. This was markedly higher than other native complement factors studied (factor B: 35.9%, C4: 45.1%, C3: 56.2%). The concentration of C9 showed a positive correlation with gestational age and was very low, 10.8% of normal adult values in cord blood and 8.3% in the patients. Fifteen percent of the neonates had C9 levels lower than 2% of adult values. The complement activation products Bb and SC5 b-9 were significantly elevated in the patients (159% and 130% of control values, respectively), indicating alternative and terminal pathway activation. In contrast, C4 bc and C3 bc levels were not increased. The maximum amount of SC5 b-9 which could be generated in the neonatal sera by cobra venom factor was highly correlated with C9 concentration (rs = 0.86, p = 0.0001) The profound C9 deficiency found in neonates is correlated with gestational age, limits the capacity to form bacteriolytic C5 b-9 (m) and may predispose for severe invasive bacterial infection. The plasma level of SC5 b-9 under normal conditions was very low, only 0.3% (0.1%-3.0%) of the values obtained after CVF activation of the same samples. Therefore, we suggest that the analysis of SC5 b-9 is applicable also in neonates, in spite of their extremely low C9 levels.
Assuntos
Proteínas do Sistema Complemento/análise , Glicoproteínas/análise , Recém-Nascido Prematuro , Bacteriemia , Proteína C-Reativa/análise , Ativação do Complemento , Complemento C3/análise , Complemento C4/análise , Complemento C9/análise , Fator B do Complemento/análise , Complexo de Ataque à Membrana do Sistema Complemento , Feminino , Sangue Fetal/química , Ruptura Prematura de Membranas Fetais , Idade Gestacional , Humanos , Imunoensaio , Recém-Nascido , Contagem de Leucócitos , Estudos Longitudinais , Masculino , Gravidez , Vitronectina/sangueRESUMO
Effects on hyperacute rejection were studied in a discordant model with the platelet GPIIb/IIIa antagonist Reopro. Pig kidneys perfused with human blood survived median 118 min in the Reopro group and 103 min in the controls (P = 0.22). Platelet and leukocyte counts decreased, whereas plasma thrombospondin and soluble as well as platelet membrane P-selectin increased significantly in both groups without significant intergroup differences. beta-Thromboglobulin and myeloperoxidase increased significantly more in the control group than in the Reopro group (P = 0.009 and P = 0.02, respectively). The classical complement pathway was substantially and similarly activated in both groups. Light and electron microscopy revealed arterial thrombi and numerous glomerular platelet aggregates in the control group in contrast to the Reopro group. In conclusion, Reopro reduced platelet aggregation, and platelet and leukocyte activation to some extent, but had no effect on complement activation and did not significantly prolong xenograft survival, even though better preservation of morphology was shown.
Assuntos
Anticorpos Monoclonais/farmacologia , Sobrevivência de Enxerto/efeitos dos fármacos , Fragmentos Fab das Imunoglobulinas/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/antagonistas & inibidores , Transplante Heterólogo/imunologia , Abciximab , Animais , Feminino , Rejeição de Enxerto , Humanos , Masculino , Contagem de Plaquetas , SuínosRESUMO
Compstatin, a newly described C3-binding peptide, inhibits complement activation by blocking C3 convertase-mediated cleavage of C3. As the complement activation is an essential part of the rejection reaction, we evaluated the ability of Compstatin to delay or prevent hyperacute rejection in an ex vivo xenograft model. Pig kidneys were perfused with fresh human blood containing either Compstatin (n=6) or a control agent (n=6). Graft survival and activation of complement, leukocytes and platelets both in the fluid-phase and in the tissue were examined. The survival of the Compstatin-perfused kidneys (median, 380 min) was significantly (P=0.0036) longer than that of the controls (median, 90 min). The classical complement pathway (C1rs-C1inhibitor and C4bc) was significantly and equally activated in both groups during the first 60 min. C3 activation products increased fivefold and terminal complement complex eightfold in the control group, but no increase occurred in the Compstatin group during this period. Immunohistochemistry showed less C3 and fibrin deposition and immune electron microscopy showed less terminal SC5b-9 complement complex deposition in the Compstatin group. A significant change in total white cells, neutrophils, myeloperoxidase, and expression of the surface activation markers CD11b (CR3) and CD35 (CR1) and CD62L (L-selectin) was observed in both groups. Leukocyte activation was lower in the Compstatin group but the difference was not statistically significant. There were no differences in platelet counts, thrombospondin, soluble P-selectin or beta-thromboglobulin between the groups. We conclude that Compstatin prolongs graft survival and suggest that it may be a useful agent for attenuating hyperacute rejection by inhibiting C3 and thus terminal complement pathway activation.
Assuntos
Proteínas Inativadoras do Complemento/farmacologia , Sobrevivência de Enxerto/efeitos dos fármacos , Peptídeos Cíclicos/farmacologia , Animais , Plaquetas/efeitos dos fármacos , Plaquetas/imunologia , Complemento C3b/metabolismo , Complemento C3b/urina , Complemento C3c/metabolismo , Complemento C3c/urina , Complexo de Ataque à Membrana do Sistema Complemento/metabolismo , Complexo de Ataque à Membrana do Sistema Complemento/urina , Feminino , Rejeição de Enxerto/prevenção & controle , Sobrevivência de Enxerto/imunologia , Humanos , Imuno-Histoquímica , Técnicas In Vitro , Transplante de Rim/imunologia , Transplante de Rim/patologia , Leucócitos/efeitos dos fármacos , Leucócitos/imunologia , Masculino , Microscopia Imunoeletrônica , Modelos Biológicos , Perfusão , Transplante HeterólogoRESUMO
The activation of complement and the release of TNF-alpha, IL-6 and IL-8 are important pathogenic factors behind organ dysfunction in sepsis. The aim of this study was to determine whether infusion of anti-TNF antibodies alters complement activation and plasma concentrations of pro-inflammatory cytokines at high doses of Escherichia coli. Six baboons received intravenously 2 x 10(9) live E. coli bacteria per kg body weight (group 1), in addition five received pretreatment with 1 mg per kg body weight anti-TNF antibodies (group 2), and seven received 5 x 10(8) live E. coli bacteria per kg body weight (group 3). Two hours after the start of infusion of the bacteria, plasma concentrations of C3 activation products, C5a and the terminal SC5b-9 complement complex were increased in groups 1 and 2 (P < 0.05), but there was no significant difference between the groups. At 2 h the levels of TNF-alpha, IL-6 and IL-8 were lower in group 2 compared with group 1 (P<0.05). In group 2 compared with group 1 the TNF-alpha concentrations were, however, higher at 4, 8 and 24 h. The explanation for this phenomenon is probably that TNF-alpha binds to the anti-TNF antibody complex and is released slowly after it has been bound. The study showed that infusion of anti-TNF antibodies reduced the concentrations of TNF-alpha, IL-6 and IL-8, without any detectable influence on complement activation.
Assuntos
Ativação do Complemento/imunologia , Infecções por Escherichia coli/imunologia , Imunoglobulinas Intravenosas/uso terapêutico , Interleucina-6/sangue , Interleucina-8/sangue , Sepse/imunologia , Fator de Necrose Tumoral alfa/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacologia , Complemento C3/imunologia , Complemento C5a/imunologia , Modelos Animais de Doenças , Infecções por Escherichia coli/sangue , Infecções por Escherichia coli/tratamento farmacológico , Imunoglobulinas Intravenosas/imunologia , Papio , Sepse/sangue , Sepse/tratamento farmacológico , Sepse/microbiologiaRESUMO
Several complement modulating effects of high-dose intravenous immunoglobulins (IVIG) have been proposed from in vitro studies and experimental animal models. However, the in vivo effects of IVIG on plasma complement in humans are yet not known. We have investigated the in vivo effects of IVIG on complement in seven women with unexplained recurrent spontaneous abortion who were without evidence of autoimmune disease. Samples were obtained before and after the very first infusion of IVIG. There was a marked increase in immunoglobulin G (IgG) from (median and range) 12.4 (9.4-15.9) to 26.8 (22.4-30.0) g/l but no change in immunoglobulin A (IgA) or immunoglobulin M (IgM). A significantly increased complement activation was demonstrated using neoepitope-specific enzyme immunoassays to the activation products C3bc (median increased from 9.8 to 31.2 AU/ml), Bb (0.66-1.66 g/ml), C5a (10.5-12.7 ng/ml), and TCC (0.81-2.19 AU/ml) (P = 0.015 for all). There were no changes in antigenic concentrations of individual complement components or regulators (C1q, C4, C3, C1-inhibitor, C4b-binding protein) and no decrease in complement haemolytic activity (classical and alternative CH50), which were all within the normal range. The classical pathway activation products C1rs/C1-inhibitor complexes, C4bc and C4d were elevated in all patients before IVIG treatment and did not change significantly during treatment. In conclusion, IVIG induced a significant activation of complement in vivo.
Assuntos
Ativação do Complemento/imunologia , Proteínas Inativadoras do Complemento , Glicoproteínas , Imunoglobulinas Intravenosas/efeitos adversos , Imunoglobulinas Intravenosas/uso terapêutico , Aborto Habitual/tratamento farmacológico , Aborto Habitual/prevenção & controle , Proteínas Sanguíneas/análise , Proteínas Sanguíneas/efeitos dos fármacos , Ativação do Complemento/efeitos dos fármacos , Proteínas Inativadoras do Complemento 1/análise , Complemento C1q/análise , Complemento C3/análise , C3 Convertase da Via Alternativa do Complemento , Complemento C3b/análise , Complemento C3c/análise , Complemento C4/análise , Complemento C5a/análise , Complexo de Ataque à Membrana do Sistema Complemento/análise , Relação Dose-Resposta Imunológica , Feminino , Humanos , Imunoglobulinas/sangue , Imunoglobulinas/efeitos dos fármacos , Imunoglobulinas Intravenosas/farmacologia , Fragmentos de Peptídeos/análise , Gravidez , Resultado da Gravidez , Receptores de Complemento/análise , Recidiva , Albumina Sérica/análise , Albumina Sérica/efeitos dos fármacosRESUMO
Sera genetically deficient in either the alpha-gamma or the beta-subunit of complement component C8 virtually lack haemolytic activity. We have studied the formation and the structural organization of the soluble terminal complement complex (TCC) assembled in these sera following activation with cobra venom factor (CVF). The TCC concentration in the activated C8alpha-gamma and C8beta-deficient samples was 0.2% and 4%, respectively, when compared with zymosan-activated normal serum. TCC was purified from the activated C8beta-deficient samples by affinity chromatography and analysed by immunoblotting and enzyme immunoassay. No C8beta was detected in one TCC preparation, while 7% of the normal level was present in the other. The level of the other terminal components, including that of C8alpha-gamma, was normal. The ability of C8alpha-gamma to promote the assembly of TCC in the presence of a limited amount of C8beta or in the apparent absence of this subunit was confirmed using purified components, by mixing C5b6 and either of the purified C8 subunits together with C7 and C9. These data show that soluble TCC can be formed in C8beta-deficient sera that contain little or no C8beta.
Assuntos
Complemento C8/deficiência , Complexo de Ataque à Membrana do Sistema Complemento/química , Complemento C5/análise , Complemento C6/análise , Complemento C7/análise , Complemento C8/análise , Complemento C9/análise , Complexo de Ataque à Membrana do Sistema Complemento/imunologia , Complexo de Ataque à Membrana do Sistema Complemento/isolamento & purificação , Humanos , Técnicas Imunoenzimáticas , Solubilidade , Fatores de TempoRESUMO
The present study was designed to investigate the consequences of isolated unilateral lung contusion on local alveolar and systemic inflammatory responses in an animal model in the pig. Isolated unilateral lung contusion was induced by bolt shot in eight mechanically ventilated animals under general anesthesia (sham: n=4). Plasma and bronchoalveolar lavage fluid were collected during a period of 8 h following lung contusion. Leukocytes, leukocyte neutral protease inhibitor (LNPI), terminal complement complex (TCC), thrombin-antithrombin-complex (TAT) as well as pulmonary microvascular permeability and surfactant function were determined. Within 30 min, lung contusion was found to cause a significant local and systemic increase in TCC and TAT concentrations and a systemic increase in LNPI concentrations. The latter was accompanied by a sequestration of leukocytes in the contused lung. Complement activation and leukocyte sequestration in the contused lung progressively increased during the investigation period. Although surfactant function decreased in the entire lung 30 min after contusion, TCC, TAT, and leukocyte sequestration was unchanged in the contralateral lung. The first indication of an involvement of the contralateral lung was obtained by an increase in leukocyte sequestration 8 h after lung contusion. Unilateral lung contusion initiates an early systemic activation of humoral and cellular defense systems. Involvement of the contralateral lung appears to be a secondary event caused by a systemic inflammatory reaction.
Assuntos
Contusões/sangue , Contusões/complicações , Inflamação/etiologia , Lesão Pulmonar , Pulmão/fisiopatologia , Animais , Antitrombina III/análise , Permeabilidade Capilar , Complexo de Ataque à Membrana do Sistema Complemento/análise , Hemodinâmica , Pulmão/irrigação sanguínea , Neutrófilos , Peptídeo Hidrolases/análise , Fosfolipídeos/análise , Fosfolipídeos/metabolismo , Proteínas Secretadas Inibidoras de Proteinases , Proteínas/análise , Alvéolos Pulmonares , Circulação Pulmonar , Troca Gasosa Pulmonar , Surfactantes Pulmonares/fisiologia , SuínosRESUMO
OBJECTIVE: Soluble complement receptor type 1 inhibits complement activation by blocking C3 and C5 convertases of the classical and alternative pathways. We evaluated the effect of soluble complement receptor type 1 on lung allograft reperfusion injury. METHODS: Left lung transplantation was performed in 13 weight-matched pigs (25 to 31 kg) after prolonged preservation (20 hours at 1 degree C). One hour after reperfusion the recipient contralateral right lung was excluded to assess graft function only. Complement activity and C3a levels were measured after reperfusion and at the end of the assessment. Extravascular lung water index, intrathoracic blood volume, and cardiac output were assessed during a 5-hour observation period. Gas exchange and hemodynamics were monitored. At the end of the 5-hour assessment period, myeloperoxidase assay and bronchoalveolar lavage were performed to assess neutrophil migration, and C5b-9 (membrane attack complex) deposits in the allograft were detected by immunohistochemistry. Two groups were studied. In group II (n = 6) recipient animals were treated with soluble complement receptor type 1 (15 mg/kg) 15 minutes before reperfusion. Group I (n = 7) served as the control group. RESULTS: Serum complement activity was completely inhibited in group II. In contrast to group I, C5b-9 complexes were not detected in group II allograft tissue samples. C3a was reduced to normal levels in group II (p = 0.00005). Extravascular lung water index was higher in group I animals throughout the assessment period (p = 0.035). No significant difference in allograft myeloperoxidase activity (p = 0.10) and polymorphonuclear leukocyte count of the bronchoalveolar lavage fluid (p = 0.057) was detected. CONCLUSION: Inhibition of the complement system by soluble complement receptor type 1 blocks local complement activation in the allograft and reduces posttransplantation reperfusion edema but does not improve hemodynamic parameters.
Assuntos
Quimiotaxia de Leucócito/fisiologia , Transplante de Pulmão , Neutrófilos/fisiologia , Edema Pulmonar/prevenção & controle , Receptores de Complemento/administração & dosagem , Traumatismo por Reperfusão/prevenção & controle , Animais , Ativação do Complemento/efeitos dos fármacos , Proteínas do Sistema Complemento/efeitos dos fármacos , Proteínas do Sistema Complemento/metabolismo , Cães , Água Extravascular Pulmonar/metabolismo , Hemodinâmica , Imuno-Histoquímica , Transplante de Pulmão/efeitos adversos , Peroxidase/metabolismo , Edema Pulmonar/etiologia , Edema Pulmonar/metabolismo , Proteínas Recombinantes , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/metabolismo , Suínos , Transplante HomólogoRESUMO
We examined alternative and classical complement activation induced by whole bacilli of Mycobacterium bovis BCG and Mycobacterium tuberculosis products. After exposure to BCG, there were higher levels of the terminal complement complex in sera from Indian tuberculosis patients than in sera from healthy controls. The addition of BCG with or without EGTA to these sera indicated that approximately 70 to 85% of the total levels of the terminal complement complex was formed by classical activation. Sera from Indian tuberculosis patients contained more antibody to lipoarabinomannan (LAM) than sera from healthy Indians. Levels of anti-LAM immunoglobulin G2 (IgG2), but not anti-LAM IgM, correlated positively with classical activation induced by BCG in the sera. By flow cytometry, deposition of C3 and terminal complement complex on bacilli incubated with normal human serum was demonstrated. The anticomplement staining was significantly reduced in the presence of EGTA and EDTA. Flow cytometry also revealed the binding of complement to BCG incubated with rabbit anti-LAM and then with factor B-depleted serum. This indicates that classical activation plays a major role in complement activation induced by mycobacteria and that anti-LAM IgG on the bacilli can mediate this response. Classical complement activation may be important for the extent of phagocytosis of M. tuberculosis by mononuclear phagocytes, which may influence the course after infection.
Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Ativação do Complemento/imunologia , Lipopolissacarídeos/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/sangue , Adolescente , Adulto , Animais , Anticorpos Antibacterianos/sangue , Feminino , Citometria de Fluxo , Humanos , Imunoglobulina M/imunologia , Masculino , Pessoa de Meia-Idade , Coelhos , Tuberculose/imunologiaRESUMO
BACKGROUND: The production in platelet concentrates (PCs) of C3 activation products (C3bc), terminal complement complex (TCC), and chemotaxins C5a, interleukin (IL)-8, tumor necrosis factor alpha (TNFalpha), and leukotriene B4 (LTB4) and the proposed reduction in concentration of the chemotaxins by white cell reduction were examined. STUDY DESIGN AND METHODS: Samples were collected from supernatants of PCs produced by apheresis (apheresis PCs) or from buffy coats (BC PCs) immediately after the production, after white cell-reduction filtration on Day 1, and after 5-day storage, and examined by enzyme immunoassays. RESULTS: Complement was activated in all PCs during storage, and the concentration of activation products was not influenced by prestorage filtration. In prestorage white cell-reduced BC PCs, only C3bc levels increased. Levels of IL-8, TNFalpha, and LTB4 increased during storage of apheresis PCs, but not in filtered units, except for LTB4. In contrast, levels of IL-8 decreased after storage of filtered BC PCs. C5a correlated significantly with IL-8, which also correlated with TNFalpha and LTB4. CONCLUSION: Both C5a and TNFalpha generation in apheresis PCs seem to induce white cell IL-8 production, which mediates cellular LTB4 release. Prestorage white cell reduction is recommended for reducing chemotactic cytokine and leukotriene levels in all PCs. Production of BC PCs is recommended to achieve less complement activation, which is not affected by filtration.
Assuntos
Plaquetas/química , Preservação de Sangue , Complemento C5a/análise , Interleucina-8/sangue , Leucotrieno B4/sangue , Ativação Plaquetária , Fator de Necrose Tumoral alfa/análise , Preservação de Sangue/efeitos adversos , Preservação de Sangue/métodos , Ativação do Complemento , Filtração , HumanosRESUMO
Pigs genetically deficient in complement factor H all develop lethal membranoproliferative glomerulonephritis (MPGN) type II characterized by massive glomerular deposits of complement, intramembranous dense deposits, and mesangial hypercellularity. To elucidate the chronological relationship between these glomerular changes, and to precisely determine the localization of glomerular complement deposits, we studied kidney specimens from factor H-deficient piglets at different ages from fetal life until terminal kidney failure had developed. Deposits of C3 and the terminal complement complex localized within the glomerular basement membrane (GBM) were present already in factor H-deficient fetuses, without concurrent intramembranous dense deposits or mesangial hypercellularity. Incipient subendothelial dense deposits containing complement appeared no earlier than four days after birth, and intramembranous dense deposits in older piglets with established MPGN type II also contained large amounts of complement as detected by immune electron microscopy. Onset of kidney failure coincided with pronounced mesangial hypercellularity and expansion, compromising glomerular capillary patency. Formation of glomerular capillary wall double contours coincided with electron microscopic evidence of laminar disintegration of intramembranous dense deposits. Complement was also deposited in the mesangial matrix, but not on glomerular cells. We conclude that all components of the alternative and terminal pathways of complement have access into the GBM and the mesangial matrix. In the absence of factor H, complement is spontaneously activated and deposited in situ in these locations resulting in dense deposit formation. It is proposed that factor H dysfunction may play an essential role even in human MPGN type II.
Assuntos
Ativação do Complemento , Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranoproliferativa/patologia , Animais , Biópsia , Peso Corporal , Fator H do Complemento/metabolismo , Proteínas do Sistema Complemento/análise , Feminino , Imunofluorescência , Glomérulos Renais/imunologia , Glomérulos Renais/patologia , Glomérulos Renais/ultraestrutura , Masculino , Microscopia Eletrônica , Microscopia Imunoeletrônica , SuínosRESUMO
Intravenous immunoglobulins (IVIG) are increasingly used for treatment of inflammatory diseases, and the modulation of complement may contribute to some of its beneficial effects. IVIG may bind C1q and activated C3 and C4, and enhance inactivation of C3b. We have previously shown that IVIG inhibited complement-mediated lysis solely via its Fc part through interaction with the classical pathway. In the present study we have investigated whole IVIG (Octagam, and Sandoglobulin) and the monomer, dimer and multimer fractions of Octagam with respect to complement activation in serum and inhibition of complement lysis of red cells. The isolated fractions were found to be stable, homogeneous (monomer, dimer or multimer) and pure (virtually only IgG). Both whole IVIG and its fractions significantly activated complement in serum and inhibited hemolysis compared with human albumin. These effects were most pronounced in the monomer, less in the multimer and least in the dimer fraction. The complement activation was shown to be mediated through the classical pathway since formation of C1rs-C1inh complexes and C4bc were increased, in contrast to Bb. Surprisingly, heat aggregation of Octagam was not followed by a corresponding increase in complement activation, as would be expected, unless it was dialysed before heating, suggesting that it is stabilized to avoid excess activation. In conclusion, the results support the hypothesis that IVIG causes a mild activation of complement in vitro. We suggest that this effect may contribute to the complement inhibitory properties of IVIG by diverting complement deposition from the target to the fluid phase.
Assuntos
Via Clássica do Complemento , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/metabolismo , Imunoglobulinas Intravenosas/imunologia , Imunoglobulinas/imunologia , Animais , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Ensaio de Atividade Hemolítica de Complemento , Eletroforese em Gel de Poliacrilamida , Eritrócitos , Calefação , Hemólise/imunologia , Humanos , Técnicas Imunoenzimáticas , Imunoglobulinas/isolamento & purificação , OvinosRESUMO
BACKGROUND: Measurement of C5a in plasma is hampered by the rapid clearance of C5a as a result of cell binding. Therefore, an assessment of whether cell-bound C5a might better reflect C5a generation in vivo is essential. METHODS: We quantified plasma and leukocyte-bound C5a in samples from patients undergoing cardiopulmonary bypass, which is known to be associated with complement activation. C3 activation products and the terminal complement complex were measured as well. RESULTS: Plasma levels of C3 activation products and the terminal complement complex increased rapidly and significantly after the onset of cardiopulmonary bypass until they reached a plateau after 30 minutes. The concentration of plasma C5a increased steadily to twice baseline at the end of bypass. The concentration of leukocyte-associated C5a increased threefold after 10 minutes of cardiopulmonary bypass, when a plateau was reached. A positive correlation was found between levels of plasma C3 activation products or terminal complement complex and plasma C5a plus cell-associated C5a but not between C3 activation products or terminal complement complex and either one of the C5a variables. CONCLUSIONS: We conclude that both plasma C5a and leukocyte-associated C5a are needed for monitoring in vivo C5a generation.
Assuntos
Ponte Cardiopulmonar , Complemento C3/análise , Complemento C5a/análise , Leucócitos/imunologia , Complemento C3/metabolismo , Complemento C5a/metabolismo , Complexo de Ataque à Membrana do Sistema Complemento/análise , Complexo de Ataque à Membrana do Sistema Complemento/metabolismo , Humanos , Contagem de Leucócitos , NeutrófilosRESUMO
In pigs a hereditary deficiency of the complement-inhibitory protein factor H consistently leads to the development of lethal membranoproliferative glomerulonephritis type II. This autosomal recessive disease has been a common cause of early losses of piglets in the Norwegian Yorkshire breed, but has not been reported in the Norwegian Landrace breed. The aim of the present work was to identify carriers of factor H deficiency and to eradicate the disease from commercial pig populations. Factor H in plasma was measured by an enzyme immunoassay. Sixteen known carriers of the disease (parents of factor H-deficient offspring) had half the level of factor H (median 110, range 87 to 156 mg/litre) recorded in 17 homozygous healthy Yorkshire pigs (median 212, range 183 to 293 mg/litre) and 20 Landrace pigs (median 227, range 200 to 255 mg/litre). Factor H analysis in 397 piglets produced by the mating of known carriers revealed an approximately 1:2:1 distribution of individuals with very low, half-normal and normal levels of factor H representing homozygous deficient, heterozygous and homozygous healthy individuals. Thus, carriers could be identified reliably by measuring the plasma concentration of factor H. Most of the population of Norwegian Yorkshire breeding pigs (490 pigs) was therefore examined, and a half-normal factor H level consistent with the carrier state was found in 13.5 per cent. These animals were prevented from breeding and since then no losses of piglets suspected of being due to factor H deficiency have been reported. No carrier was identified among 102 Norwegian Landrace boars, almost excluding the existence of factor H deficiency in this breed.
Assuntos
Fator H do Complemento/deficiência , Glomerulonefrite Membranoproliferativa/veterinária , Doenças dos Suínos/genética , Doenças dos Suínos/prevenção & controle , Envelhecimento/sangue , Animais , Cruzamento , Fator H do Complemento/análise , Fator H do Complemento/genética , Triagem de Portadores Genéticos/métodos , Testes Genéticos/métodos , Testes Genéticos/veterinária , Glomerulonefrite Membranoproliferativa/genética , Glomerulonefrite Membranoproliferativa/prevenção & controle , Homozigoto , Técnicas Imunoenzimáticas/veterinária , Noruega/epidemiologia , Suínos , Doenças dos Suínos/epidemiologiaRESUMO
BACKGROUND: Acquired deficiencies of certain complement proteins and impaired opsonisation activity have been implicated in the pathogenesis of the increased susceptibility to infections of patients with alcoholic cirrhosis. METHODS: Serum concentrations of C3 and C4, plasma concentrations of C3bc, C9, and the terminal C5b-9 complement complex (TCC), and haemolytic complement activity (classic and alternative pathway) of serum, and serum opsonic activity were determined in 46 patients with compensated alcoholic cirrhosis, 31 who were decompensated, and in 15 healthy subjects. After 19 months (median) the investigated variables were analysed for their use in prognosis of recurrent infections and survival. RESULTS: C3 and C4 concentrations and the haemolytic complement activity of the alternative pathway were decreased in decompensated cirrhotic patients compared with controls (p < 0.01). Univariate analysis (log rank test) showed that low concentrations (< or = lower quartile) of C3 (p < 0.001) and C3bc (p < 0.05), haemolytic complement activity of the alternative pathway (p < 0.01) and classic pathway (p < 0.05), and decompensated cirrhosis (p < 0.001) were associated with an increased risk of infection and increased mortality. Multivariate (Cox) analysis showed that low C3 concentrations and decompensation of cirrhosis were significant predictors of infections and mortality (p < 0.02). CONCLUSIONS: Low serum C3 concentrations and decreased haemolytic complement function predisposes to infection and increased mortality in patients with alcoholic cirrhosis.
Assuntos
Infecções Bacterianas/imunologia , Ativação do Complemento , Complemento C3/deficiência , Cirrose Hepática Alcoólica/imunologia , Adulto , Idoso , Infecções Bacterianas/complicações , Complemento C3/análise , Complemento C4/análise , Complemento C9/análise , Feminino , Humanos , Cirrose Hepática Alcoólica/complicações , Cirrose Hepática Alcoólica/microbiologia , Cirrose Hepática Alcoólica/mortalidade , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Fatores de RiscoRESUMO
BACKGROUND: The inflammatory response induced by cardiopulmonary bypass can result in severe organ dysfunction in some patients. This postperfusion response is caused mainly by contact between blood and the foreign surface of the cardiopulmonary bypass equipment and includes adhesion of leukocytes to vascular endothelium, which precedes a series of events that mediate inflammatory damage to tissues. METHODS: Low-risk patients accepted for coronary artery bypass grafting were randomized to operation with the cardiopulmonary bypass surface either completely heparin coated (Duraflo II) or uncoated. There were 12 patients in each group. Blood plasma sampled during cardiopulmonary bypass was analyzed for complement activation (C3bc and terminal SC5b-9 complement complex) and neutrophil activation (lactoferrin and myeloperoxidase). In addition, neutrophils, monocytes, and platelets were counted, and the expression of surface markers on the neutrophils and monocytes (complement receptor [CR] 1, CR3, CR4, and L-selectin) and on the platelets (P-selectin and CD41) was quantified with flow cytometry. RESULTS: Clinical and surgical results were similar in both groups. In the group with the heparin-coated surface, the formation of the terminal SC5b-9 complement complex was significantly reduced, and the counts of circulating leukocytes and platelets were significantly less reduced initially but were higher at the end of cardiopulmonary bypass compared with baseline. Also, the expression of CR1, CR3, and CR4 was significantly less upregulated and the L-selectin, significantly less downregulated on monocytes and neutrophils. CONCLUSIONS: We conclude that heparin coating reduces complement activation and attenuates the leukocyte integrin and selectin response that occurs when uncoated circuits are used.