RESUMO
The complement system, an important part of the innate system, is known to play a central role in many immune mediated kidney diseases. All parts of the complement system including the classical, alternative, and mannose-binding lectin pathways have been implicated in complement-mediated kidney injury. Although complement components are thought to be mainly synthesized in the liver and activated in the circulation, emerging data suggest that complement is synthesized and activated inside the kidney leading to direct injury. Urinary complement biomarkers are likely a better reflection of inflammation within the kidneys as compared to traditional serum complement biomarkers which may be influenced by systemic inflammation. In addition, urinary complement biomarkers have the advantage of being non-invasive and easily accessible. With the rise of therapies targeting the complement pathways, there is a critical need to better understand the role of complement in kidney diseases and to develop reliable and non-invasive biomarkers to assess disease activity, predict treatment response and guide therapeutic interventions. In this review, we summarized the current knowledge on urinary complement biomarkers of kidney diseases due to immune complex deposition (lupus nephritis, primary membranous nephropathy, IgA nephropathy) and due to activation of the alternative pathway (C3 glomerulopathy, thrombotic microangiography, ANCA-associated vasculitis). We also address the limitations of current research and propose future directions for the discovery of urinary complement biomarkers.
Assuntos
Biomarcadores , Proteínas do Sistema Complemento , Nefropatias , Humanos , Biomarcadores/urina , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/urina , Proteínas do Sistema Complemento/metabolismo , Nefropatias/urina , Nefropatias/imunologia , Nefropatias/diagnóstico , Animais , Ativação do ComplementoRESUMO
Introduction: Crescents, especially those found at a percentage greater than 50%, are often associated with rapid progression of kidney disease in IgA nephropathy (IgAN). The mechanism of crescents forming in IgAN is still unclear. In this study, we aimed to evaluate whether excess complement activation participates in the formation of crescents in IgAN. Methods: One hundred IgAN patients with various proportions of crescents-24 with 1%-24%, 27 with 25%-49%, 21 with 50%-74% 12 with more than 75%, and 16 without crescents-were included. Urinary concentrations of mannose-binding lectin (MBL), Bb, C4d, C3a, C5a, and soluble C5b-9 (sC5b-9) were measured at the time of biopsy. Receiver operating characteristic (ROC) curves were performed to evaluate predictive ability of renal survival for urine complement activation. In addition, historical C4d, C5b-9, and C3d were stained by immunohistochemistry. Results: IgAN patients with more than 50% crescent formation showed higher complement activation levels than the other patients (urinary C3a/creatinine (C3a/Cr): 6.7295 ng/mg, interquartile range (IQR) 1.4652-62.1086 ng/mg vs. 0.1055 ng/mg, IQR 0-1.4089 ng/mg; urinary C5a/Cr: 15.6202 ng/mg, 4.3127-66.7347 ng/mg vs. 0.3280 ng/mg, IQR 0.0859-2.4439 ng/mg; urinary sC5b-9/Cr: 98.6357 ng/mg, 8.8058-1,087.4578 ng/mg vs. 1.4262 ng/mg, 0.0916-11.0858 ng/mg, all p-values <0.001). The levels of urinary MBL and C4d representing lectin complement pathway showed a linear association with the proportion of crescents (r = 0.457 and 0.562, respectively, both p-values <0.001). Combined urine complement products could increase the predictive ability compared with crescents alone from 0.904 to 0.944 (p = 0.062) with borderline significance. Moreover, the glomerular C4d deposition rate elevated with the increase of proportions of crescents. Conclusion: Excess complement activation may be involved in the formation of crescents, especially diffuse crescent formation, in patients with IgAN. Urinary C4d correlated with the proportion of crescents and was a potential biomarker for disease monitoring in crescentic IgAN.
Assuntos
Ativação do Complemento , Proteínas do Sistema Complemento/urina , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Índice de Gravidade de Doença , Adulto , Biópsia , Feminino , Glomerulonefrite por IGA/sangue , Glomerulonefrite por IGA/urina , Humanos , Imuno-Histoquímica/métodos , Glomérulos Renais/patologia , Masculino , Lectina de Ligação a Manose/urina , Pessoa de Meia-Idade , Prognóstico , Adulto JovemRESUMO
The kidney is one of the main organs affected by the autoimmune disease systemic lupus erythematosus. Lupus nephritis (LN) concerns 30-60% of adult SLE patients and it is significantly associated with an increase in the morbidity and mortality. The definitive diagnosis of LN can only be achieved by histological analysis of renal biopsies, but the invasiveness of this technique is an obstacle for early diagnosis of renal involvement and a proper follow-up of LN patients under treatment. The use of urine for the discovery of non-invasive biomarkers for renal disease in SLE patients is an attractive alternative to repeated renal biopsies, as several studies have described surrogate urinary cells or analytes reflecting the inflammatory state of the kidney, and/or the severity of the disease. Herein, we review the main findings in the field of urine immune-related biomarkers for LN patients, and discuss their prognostic and diagnostic value. This manuscript is focused on the complement system, antibodies and autoantibodies, chemokines, cytokines, and leukocytes, as they are the main effectors of LN pathogenesis.
Assuntos
Biomarcadores/urina , Nefrite Lúpica/imunologia , Nefrite Lúpica/urina , Autoanticorpos/imunologia , Autoanticorpos/urina , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/urina , Diagnóstico Precoce , Humanos , Cadeias Leves de Imunoglobulina/imunologia , Cadeias Leves de Imunoglobulina/urina , Inflamação/imunologia , Inflamação/urina , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/urina , Nefrite Lúpica/diagnóstico , PrognósticoRESUMO
PURPOSE: To investigate the association between urinary complement proteins and renal outcome in biopsy-proven diabetic nephropathy (DN). METHODS: Untargeted proteomic and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional analyses and targeted proteomic analysis using parallel reaction-monitoring (PRM)-mass spectrometry was performed to determine the abundance of urinary complement proteins in healthy controls, type 2 diabetes mellitus (T2DM) patients, and patients with T2DM and biopsy-proven DN. The abundance of each urinary complement protein was individually included in Cox proportional hazards models for predicting progression to end-stage renal disease (ESRD). RESULTS: Untargeted proteomic and functional analysis using the KEGG showed that differentially expressed urinary proteins were primarily associated with the complement and coagulation cascades. Subsequent urinary complement proteins quantification using PRM showed that urinary abundances of C3, C9, and complement factor H (CFAH) correlated negatively with annual estimated glomerular filtration rate (eGFR) decline, while urinary abundances of C5, decay-accelerating factor (DAF), and CD59 correlated positively with annual rate of eGFR decline. Furthermore, higher urinary abundance of CFAH and lower urinary abundance of DAF were independently associated with greater risk of progression to ESRD. Urinary abundance of CFAH and DAF had a larger area under the curve (AUC) than that of eGFR, proteinuria, or any pathological parameter. Moreover, the model that included CFAH or DAF had a larger AUC than that with only clinical or pathological parameters. CONCLUSION: Urinary abundance of complement proteins was significantly associated with ESRD in patients with T2DM and biopsy-proven DN, indicating that therapeutically targeting the complement pathway may alleviate progression of DN.
Assuntos
Proteínas do Sistema Complemento , Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas , Falência Renal Crônica , Rim/patologia , Biópsia/métodos , Proteínas do Sistema Complemento/metabolismo , Proteínas do Sistema Complemento/urina , Correlação de Dados , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/fisiopatologia , Progressão da Doença , Descoberta de Drogas , Feminino , Humanos , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/metabolismo , Falência Renal Crônica/prevenção & controle , Testes de Função Renal/métodos , Masculino , Pessoa de Meia-Idade , Prognóstico , Modelos de Riscos Proporcionais , Proteinúria/diagnóstico , Proteinúria/etiologia , Proteômica/métodos , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Studies on adriamycin mice model suggest complement system is activated and together with IgM contributes to the glomerular injury of primary focal segmental glomerulosclerosis (FSGS). We recently reported primary FSGS patients with IgM and C3 deposition showed unfavorable therapeutic responses and worse renal outcomes. Here we examined the plasma and urinary complement profile of patients with primary FSGS, aiming to investigate the complement participation in FSGS pathogenesis. METHODS: Seventy patients with biopsy-proven primary FSGS were enrolled. The plasma and urinary levels of C3a, C5a, soluble C5b-9, C4d, C1q, MBL, and Bb were determined by commercial ELISA kits. RESULTS: The levels of C3a, C5a and C5b-9 in plasma and urine of FSGS patients were significantly higher than those in normal controls. The plasma and urinary levels of C5b-9 were positively correlated with urinary protein, renal dysfunction and interstitial fibrosis. The plasma C5a levels were positively correlated with the proportion of segmental sclerotic glomeruli. The urinary levels of Bb were elevated, positively correlated with C3a and C5b-9 levels, renal dysfunction, and interstitial fibrosis. The plasma C1q level was significantly decreased, and negatively correlated with urinary protein excretion. Urinary Bb level was a risk factor for no remission (HR = 3.348, 95% CI 1.264-8.870, P = 0.015) and ESRD (HR = 2.323, 95% CI 1.222-4.418, P = 0.010). CONCLUSION: In conclusion, our results identified the systemic activation of complement in human primary FSGS, possibly via the classical and alternative pathway. The activation of complement system was partly associated with the clinical manifestations, kidney pathological damage, and renal outcomes.
Assuntos
Ativação do Complemento/imunologia , Proteínas do Sistema Complemento , Glomerulosclerose Segmentar e Focal/imunologia , Glomérulos Renais , Adulto , Biomarcadores/sangue , Biomarcadores/urina , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/urina , Feminino , Humanos , Glomérulos Renais/imunologia , Glomérulos Renais/lesões , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
OBJECTIVE: To investigate the association between complement and inflammatory biomarkers with diabetic nephropathy (DN) in type 2 diabetes mellitus (T2DM). METHODS: Plasma and urinary complement and inflammatory biomarkers were measured in 115 T2DM patients assigned to one of two groups: with DN (n=48) and without DN (n=67). RESULTS: The plasma and urinary levels of C3a, C4d, C5a, sC5b-9 and MBL (mannan-binding lectin) were significantly higher in T2DM patients with DN compared to T2DM patients without DN. The plasma levels of IL-10 and INF-γ, as well as the urinary levels of INF-γ and TNF-α in T2DM patients with DN, were significantly higher than T2DM patients without DN. Both urinary MBL and INF-γ were independent risk factors for DN within T2DM patients (OR, 2.35 (95% CI 2.28-2.64) and 1.17 (95% CI 1.15-1.18); P=0.000 and 0.016, respectively). The area under the receiver-operating-characteristic-curve for urinary MBL was 0.89, with sensitivity 91% and specificity 83% for DN. The area under the receiver-operating-characteristic-curve for INF-γ was 0.84, with sensitivity 86% and specificity 79% based on cutoff values of 1.42 ng/mg and 5.15 pg/mg, respectively. CONCLUSION: This study suggests that urinary INF-γ and MBL levels are independent risk factors with a high predictive power for DN in T2DM patients.
Assuntos
Biomarcadores/sangue , Proteínas do Sistema Complemento/metabolismo , Nefropatias Diabéticas/sangue , Inflamação/sangue , Adulto , Idoso , Área Sob a Curva , Biomarcadores/urina , Proteínas do Sistema Complemento/urina , Nefropatias Diabéticas/urina , Feminino , Humanos , Inflamação/urina , Mediadores da Inflamação/sangue , Mediadores da Inflamação/urina , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Curva ROCRESUMO
BACKGROUND: Complement activation plays a substantial role in the pathogenesis of primary membranous nephropathy (pMN). C5b-9, C3c, MBL, and factor B have been documented in the subepithelial immune deposits. However, the changing of complement activation products in circulation and urine is not clear. METHODS: We measured the circulating and urinary levels of C1q, MBL, C4d, Bb, properdin, C3a, C5a, and sC5b-9, in 134 patients with biopsy-proven pMN, by enzyme-linked immunosorbent assay. All the plasma values were corrected by eGFR and all the urinary values were corrected by urinary creatinine and urinary protein excretion. Anti-PLA2R antibodies were measured in all patients. RESULTS: The plasma complement activation products were elevated both in the patients with and without anti-PLA2R antibodies. C3a levels were remarkably increased in the circulation and urine, much higher than the elevated levels of C5a. C5b-9 was in normal range in plasma, but significantly higher in urine. The urinary C5a had a positive correlation with anti-PLA2R antibody levels and urinary protein. The plasma level of C4d was elevated, but C1q and MBL were comparable to healthy controls. Positive correlations were observed between plasma C4d/MBL and urinary protein, only in the patients with positive anti-PLA2R antibodies but not in those without. The plasma level of Bb was elevated and had positive correlation with urinary protein only in the patients without anti-PLA2R antibodies. CONCLUSION: Complement activation products were remarkable increased in pMN and may serve as sensitive biomarkers of disease activity. The complement may be activated through lectin pathway with the existence of anti-PLA2R antibodies, while through alternative pathway in the absence of antibody.
Assuntos
Ativação do Complemento , Proteínas do Sistema Complemento/análise , Glomerulonefrite Membranosa/sangue , Glomerulonefrite Membranosa/urina , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Complemento C1q/análise , Complemento C1q/urina , Complemento C3a/análise , Complemento C3a/urina , Complemento C4/análise , Complemento C4/urina , Complemento C5a/análise , Complemento C5a/urina , Fator B do Complemento/análise , Fator B do Complemento/urina , Complexo de Ataque à Membrana do Sistema Complemento/análise , Complexo de Ataque à Membrana do Sistema Complemento/urina , Proteínas do Sistema Complemento/urina , Creatinina/sangue , Creatinina/urina , Feminino , Glomerulonefrite Membranosa/imunologia , Glomerulonefrite Membranosa/terapia , Humanos , Masculino , Lectina de Ligação a Manose/sangue , Lectina de Ligação a Manose/urina , Pessoa de Meia-Idade , Properdina/análise , Properdina/urina , Receptores da Fosfolipase A2/análise , Receptores da Fosfolipase A2/sangue , Receptores da Fosfolipase A2/imunologia , Análise de Regressão , Estatísticas não Paramétricas , Adulto JovemRESUMO
OBJECTIVE: We examined the association of urine complement proteins with progression to end-stage renal disease (ESRD) or death in people with type 2 diabetes and proteinuric diabetic kidney disease (DKD). RESEARCH DESIGN AND METHODS: Using targeted mass spectrometry, we quantified urinary abundance of 12 complement proteins in a predominantly Mexican American cohort with type 2 diabetes and proteinuric DKD (n = 141). The association of urine complement proteins with progression to ESRD or death was evaluated using time-to-event analyses. RESULTS: At baseline, median estimated glomerular filtration rate (eGFR) was 54 mL/min/1.73 m2 and urine protein-to-creatinine ratio 2.6 g/g. Sixty-seven participants developed ESRD or died, of whom 39 progressed to ESRD over a median of 3.1 years and 40 died over a median 3.6 years. Higher urine CD59, an inhibitor of terminal complement complex formation, was associated with a lower risk of ESRD (hazard ratio [HR] [95% CI per doubling] 0.50 [0.29-0.87]) and death (HR [95% CI] 0.56 [0.34-0.93]), after adjustment for demographic and clinical covariates, including baseline eGFR and proteinuria. Higher urine complement components 4 and 8 were associated with lower risk of death (HR [95% CI] 0.57 [0.41-0.79] and 0.66 [0.44-0.97], respectively); higher urine factor H-related protein 2, a positive regulator of the alternative complement pathway, was associated with greater risk of death (HR [95% CI] 1.61 [1.05-2.48]) in fully adjusted models. CONCLUSIONS: In a largely Mexican American cohort with type 2 diabetes and proteinuric DKD, urine abundance of several complement and complement regulatory proteins was strongly associated with progression to ESRD and death.
Assuntos
Proteínas do Sistema Complemento/urina , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/mortalidade , Diabetes Mellitus Tipo 2/urina , Nefropatias Diabéticas/diagnóstico , Falência Renal Crônica/diagnóstico , Idoso , Estudos Transversais , Diabetes Mellitus Tipo 2/etnologia , Nefropatias Diabéticas/etnologia , Nefropatias Diabéticas/mortalidade , Nefropatias Diabéticas/urina , Progressão da Doença , Feminino , Taxa de Filtração Glomerular , Humanos , Falência Renal Crônica/etnologia , Falência Renal Crônica/mortalidade , Falência Renal Crônica/urina , Masculino , Espectrometria de Massas , Americanos Mexicanos , Pessoa de Meia-Idade , Prognóstico , Proteinúria/diagnóstico , Proteinúria/etnologia , Proteinúria/mortalidade , Proteinúria/patologia , Fatores de Risco , Análise de SobrevidaRESUMO
AIM: To compare the clinical manifestations membranoproliferative glomerulonephritis (MPGN) in its idiopathic variant, lupus nephritis (LN), and C3 glomerulopathy (C3-GP), by comparing them with changes in the complement system. SUBJECTS AND METHODS: The clinic of nephrology followed up 42 patients with different types of MPGN in 2013 to 2015. The study included 35 patients divided into 3 groups: 1) 8 patients with C3-GP, 2) 13 with idiopathic MPGN; 3) 14 with Class IV LN. The investigators studied the blood and urine levels of components and markers for activation of the classical and alternative pathways (C3 and C4, С3а, C5a, CFH, CFB, and CFD) of the terminal complement complex (TCC). RESULTS: The detection rate of C3-GP was 19%. The patients with C3-GP were noted to have the lowest blood concentration of S3 and the highest urinary level of С3а, C5a, TCC, CFH, CFB, and CFD. C3 nephritic factor was detected in 2 patients from the C3-GP (dense deposit disease) group. CONCLUSION: Alternative complement pathway dysregulation caused by genetic or autoimmune factors plays a leading role in the pathogenesis of C3-GP.
Assuntos
Complemento C3/metabolismo , Proteínas do Sistema Complemento/metabolismo , Glomerulonefrite Membranoproliferativa , Nefrite Lúpica , Adulto , Complemento C3/urina , Proteínas do Sistema Complemento/urina , Feminino , Glomerulonefrite Membranoproliferativa/sangue , Glomerulonefrite Membranoproliferativa/urina , Humanos , Nefrite Lúpica/sangue , Nefrite Lúpica/urina , MasculinoRESUMO
BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat inflammatory pain in humans and animals. An overdose of an NSAID is nephrotoxic and can lead to acute kidney injury (AKI). Complement activation occurs in several types of renal disorders with proteinuria. The aim of this study was to investigate whether complement system becomes activated in kidneys after a high dose of NSAID. Kidney tissue and urine samples were collected from six sheep with ketoprofen-induced AKI and from six healthy control sheep. The localization of complement proteins in kidney tissue was carried out using immunohistochemical stainings, and excretion of C3 was tested by immunoblotting. RESULTS: The complement system was found to become activated in the kidney tissue as demonstrated by positive immunostaining for C1q, C3c, C4c, C5, C9 and factor H and by Western blotting analysis of C3 activation products in urine samples in sheep with AKI. CONCLUSIONS: Our results thus suggest that the alternative complement pathway is activated, and it may contribute to the acute tubular injury seen in the kidneys of NSAID-induced AKI sheep. Inhibition of complement activation may serve as potential therapeutic target for intervention in drug-induced AKI.
Assuntos
Injúria Renal Aguda/fisiopatologia , Anti-Inflamatórios não Esteroides/efeitos adversos , Proteínas do Sistema Complemento/metabolismo , Cetoprofeno/efeitos adversos , Doenças dos Ovinos/fisiopatologia , Injúria Renal Aguda/induzido quimicamente , Animais , Anti-Inflamatórios não Esteroides/urina , Western Blotting/veterinária , Proteínas do Sistema Complemento/urina , Feminino , Imuno-Histoquímica/veterinária , Cetoprofeno/urina , Rim/lesões , Ovinos , Doenças dos Ovinos/induzido quimicamenteRESUMO
OBJECTIVES: The complement system is involved in many immune complex-mediated kidney diseases, yet its role in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD) has not been examined in detail. METHODS AND RESULTS: Screening of the glycoproteome of urine samples from ADPKD patients revealed that levels of complement factor B (CFB), serpin peptidase inhibitor, complement component 1 inhibitor (SERPING1) and complement component 9 (C9) increased, whereas complement component 1, r subcomponent-like (C1RL), CD55 and CD59 levels decreased with disease progression. Immunostaining and Western blot analysis confirmed the enhanced expression of CFB and C9 in cystic kidneys from ADPKD patients. Immunostaining also showed that the expressions of CFB and C9 in renal biopsy tissues from patients with other types of chronic kidney disease were lower than in tissues from ADPKD patients. The effect of the complement inhibitor rosmarinic acid (RMA) was evaluated in Pkd1(-/-) mice and Han:SPRD Cy/+ rats. Compared with vehicle-treated Pkd1(-/-) animals, RMA-treated mice had significantly lower serum creatinine (-50%) and blood urea nitrogen (-78%) levels, two kidneys/body weight ratio (-60%) and renal cystic index (-60%). Similar results were found in Cy/+ rats. Lower numbers of Ki67-positive nuclei and inflammatory cells and reduced fibrosis were observed in both animal models upon treatment with RMA. CONCLUSIONS: These results suggest that excessive activation of the alternative complement pathway is associated with ADPKD progression, probably mediated by cyst-lining epithelial cell proliferation, tubulointerstitial inflammatory cell infiltration and fibrosis. Targeting the complement system might represent a new therapeutic strategy for ADPKD.
Assuntos
Via Alternativa do Complemento , Rim Policístico Autossômico Dominante/imunologia , Adulto , Animais , Proliferação de Células , Complemento C3/metabolismo , Complemento C4/metabolismo , Complemento C9/metabolismo , Fator B do Complemento/metabolismo , Via Alternativa do Complemento/efeitos dos fármacos , Proteínas do Sistema Complemento/urina , Progressão da Doença , Células Epiteliais/metabolismo , Fibrose , Humanos , Rim/metabolismo , Rim/patologia , Camundongos Knockout , Pessoa de Meia-Idade , Rim Policístico Autossômico Dominante/patologia , Rim Policístico Autossômico Dominante/urina , RatosRESUMO
BACKGROUND AND OBJECTIVES: Previous study revealed that complement activation products of the alternative pathway could be detected in renal specimens of human ANCA-associated vasculitis. The current study aimed to investigate the clinical and pathologic significance of complement activation products in the urine and kidneys of patients with ANCA-associated vasculitis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Renal biopsy specimens from 29 patients with ANCA-associated vasculitis diagnosed at Peking University First Hospital from January of 2008 to December of 2010 were randomly collected. Urine samples from 27 of 29 patients in active stage and 22 ANCA-associated vasculitis patients in complete remission who were independent of the above-mentioned 29 patients were collected. Urine samples from 28 patients with lupus nephritis and 25 healthy individuals were also collected. The renal deposition of Bb, C3d, and C5b-9 were detected by immunohistochemistry. The urinary levels of Bb, C3a, C5a, and soluble C5b-9 were determined by ELISA. RESULTS: The deposition, measured by the mean optical density of Bb, which is an alternative complement pathway marker, in glomeruli correlated with the proportion of total crescents (r=0.50, P=0.006), the extent of interstitial infiltrate (r=0.59, P=0.001), interstitial fibrosis (r=0.45, P=0.01), and tubular atrophy (r=0.55, P=0.002), whereas it correlated inversely with the proportion of normal glomeruli (r=-0.49, P=0.008). The urinary levels of Bb, C3a, C5a, and soluble C5b-9 were all significantly higher in active compared with remission stage. The urinary levels of Bb in patients with active ANCA-associated vasculitis correlated with the serum creatinine (r=0.56, P=0.002) and correlated inversely with the proportion of normal glomeruli in renal specimens (r=-0.49, P=0.009). CONCLUSIONS: The present study provides additional evidence that complement activation through the alternative pathway occurred in the development of ANCA-associated vasculitis. The renal deposition of Bb and urinary Bb levels were associated with the severity of renal injury.
Assuntos
Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/imunologia , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/urina , Anticorpos Anticitoplasma de Neutrófilos/análise , Via Alternativa do Complemento , Proteínas do Sistema Complemento/urina , Glomerulonefrite/imunologia , Glomerulonefrite/urina , Rim/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/sangue , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/patologia , Biomarcadores/sangue , Biomarcadores/urina , Biópsia , Estudos de Casos e Controles , China , Fator B do Complemento/urina , Creatinina/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Glomerulonefrite/sangue , Glomerulonefrite/patologia , Hospitais Universitários , Humanos , Imuno-Histoquímica , Rim/patologia , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Urinálise/métodos , Adulto JovemRESUMO
PURPOSE: Linear or granular deposition of complement 3 (C3) along glomerular basement membrane (GBM) is generally revealed in kidneys of human anti-GBM disease. However, the mechanism of complement activation and its association with clinical features and outcomes are less clear. METHODS: We measured the plasma and urinary levels of complement components, C1q, mannose-binding lectin (MBL), factor B (Ba), C3, C3a, C4, C4a, C5, C5a and soluble C5b-9 (SC5b-9), using ELISA in 20 patients with renal biopsy proven anti-GBM disease. RESULTS: The end product of complement activation, SC5b-9, was elevated both in plasma and urine. The levels of C3 and C4 were normal in plasma, while elevated in urine. The levels of C5a and SC5b-9 were increased in plasma from 15% and 30% patients respectively, while they were raised in urine from almost all patients (100% and 92%). The levels of plasma SC5b-9 and urinary C5a were positively correlated with the serum creatinine at presentation (r=0.56, P=0.01; r=0.68, P=0.02, respectively) and the percentage of crescents in glomeruli (r=0.60, P=0.005; r=0.75, P=0.005, respectively). The plasma level of SC5b-9 was further identified as the predictor for renal failure during follow up (HR, 1.46; 95% CI, 1.12-1.90; P=0.005). CONCLUSION: Complement cascade goes to the end in human anti-GBM disease and resides mainly in kidney. It plays pathogenic role in renal injury, by the possible proinflammatory effect of C5a and/or cell lysis effect of C5b-9. C5a and C5b-9 may be useful in clinical monitoring and predicting.
Assuntos
Doença Antimembrana Basal Glomerular/imunologia , Doença Antimembrana Basal Glomerular/patologia , Ativação do Complemento/imunologia , Rim/imunologia , Rim/lesões , Adolescente , Adulto , Idoso , Doença Antimembrana Basal Glomerular/fisiopatologia , Proteínas do Sistema Complemento/metabolismo , Proteínas do Sistema Complemento/urina , Feminino , Humanos , Rim/patologia , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
AIM: To investigate the involvement of complement activation and apoptosis in the pathogenesis of membranous glomerulonephritis by determining the concentrations of apoptosis-associated 180 bp nucleosomes and complement activation products SC5b-9 and C3d/dg in the urine of patients with membranous glomerulonephritis. METHODS: Morning urine was taken from 15 patients with immunohistologically established membranous glomerulonephritis. Apoptosis-associated 180 bp nucleosomes, complement activation products SC5b-9, C3d/dg, and immune complexes CIC-C3d, CIC-IgA, and CIC-IgG were detected in the urine samples by using antigen-specific enzyme-linked immunosorbent assay. RESULTS: Concentrations of measured parameters were expressed in units of standard deviation, ie, relatively to the average concentrations measured in healthy subjects. We found drastically increased concentrations of apoptosis-associated 180 bp nucleosomes (13.71+/-14.97; p=0.047), complement activation products SC5b-9 (197.07+/-134.88; p=0.003) and C3d/dg (38.70+/-43.35; p=0.048), and immune complexes CIC-C3d (11.01+/-13.39; p=0.74), CIC-IgA (7.93+/-4.38; p=0.001), and CIC-IgG (20.56+/-10.87; p=0.001) in the urine of patients with an active form of membranous glomerulonephritis. All studied molecules were absent, or present in very low concentrations, in healthy subjects and patients with membranous glomerulonephritis in remission. The mean differences between healthy controls and patients with the active disease were statistically significant in all parameters, except CIC-C3d. CONCLUSIONS: There is an association of complement activation and apoptosis with membranous glomerulonephritis. Correlation analysis suggests that the excretion of apoptosis-associated 180 bp nucleosomes, SC5b-9, C3d/dg, and immune complexes containing IgA and IgG in the urine of patients with active membranous glomerulonephritis does not depend solely on the passive transport together with other proteins, but is probably an independent active process.
Assuntos
Complexo Antígeno-Anticorpo/urina , Complemento C3b/urina , Proteínas do Sistema Complemento/urina , Glomerulonefrite Membranosa/urina , Glicoproteínas/urina , Fragmentos de Peptídeos/urina , Adulto , Idoso , Apoptose , Divisão Celular , Ativação do Complemento , Complexo de Ataque à Membrana do Sistema Complemento , Feminino , Humanos , Imunoglobulina A/urina , Imunoglobulina G/urina , Masculino , Pessoa de Meia-Idade , Proteínas Repressoras/urinaAssuntos
Proteínas do Sistema Complemento/urina , Túbulos Renais/fisiopatologia , Nefrite Intersticial/fisiopatologia , Proteinúria/fisiopatologia , Animais , Antígenos CD , Permeabilidade Capilar , Ativação do Complemento , Complexo de Ataque à Membrana do Sistema Complemento/urina , Proteínas do Sistema Complemento/biossíntese , Citocinas/biossíntese , Progressão da Doença , Fibrose , Mesângio Glomerular/fisiopatologia , Humanos , Túbulos Renais/metabolismo , Túbulos Renais Proximais/fisiopatologia , Camundongos , Modelos Biológicos , Receptor da Anafilatoxina C5a , Receptores de Complemento/deficiênciaRESUMO
Protectin (CD59) is a low molecular weight glycophosphoinositol-anchored inhibitor of the membrane attack complex of complement (MAC) that is present, for example, on the membranes of endothelial cells and on epithelial cells of glomeruli and distal tubuli. To examine for the possibility that CD59 becomes detached from cell surfaces following cell injury, this study evaluated renal excretion of CD59 in patients with idiopathic membranous glomerulonephritis (MGN; N = 21), diabetic nephropathy (DNP; N = 15) and in healthy control subjects (N = 13). CD59 in human urine was quantitated by a competitive solid-phase radioimmunoassay having approximately 13 kDa soluble urinary CD59 as a standard. Immunofluorescence microscopy demonstrated a decreased expression of CD59 in the glomeruli of MGN patients. Using a Triton X-114 phase separation method 91 to 97% of urinary CD59 was found to be in a soluble form without anchor-associated phospholipid. The mean (+/- SEM) level of urinary CD59 was 5.6 +/- 0.2 micrograms/ml in MGN patients, 3.7 +/- 0.4 micrograms/ml in healthy controls (P < 0.001) and 2.6 +/- 0.1 in DNP patients (P < 0.001). When related to urinary creatinine (UCr) the corresponding values were 11.9 +/- 5.6, 4.8 +/- 0.3 (P = 0.021) and 4.4 +/- 0.2 (P < 0.002), respectively. The amount of CD59 in urine correlated with the urinary excretion of soluble terminal complement complexes, SC5b-9 (r = 0.594, P < 0.006) in MGN patients. The excretion of CD59 also correlated with the excretion of the inflammatory mediator IL-1 beta (r = 0.671, P = 0.001) but not with TNF-alpha (r = 0.314, P = 0.178). No correlation of CD59 excretion was observed with duration of the disease level of proteinuria, serum albumin concentration or serum creatinine level. Based on these findings we speculate that the increased excretion of CD59 into urine in MGN patients is due to complement activation and inflammation induced shedding of CD59 from glomerular cells.
Assuntos
Antígenos CD/urina , Proteínas do Sistema Complemento/urina , Citocinas/urina , Glomerulonefrite Membranosa/urina , Glicoproteínas/urina , Glicoproteínas de Membrana/urina , Adulto , Análise de Variância , Antígenos CD59 , Complexo de Ataque à Membrana do Sistema Complemento , Progressão da Doença , Seguimentos , Humanos , Rim/metabolismo , Microscopia de Fluorescência , Pessoa de Meia-Idade , RadioimunoensaioRESUMO
Increasing evidence exists that inducible adhesion molecules are involved in cell-mediated allograft rejection. In addition, complement activation during rejection has been described. This study investigated, whether specific molecules derived from either pathway are excreted into urine during rejection and whether they can provide useful diagnostic tools for the monitoring of renal transplant recipients. Urinary concentrations of soluble adhesion molecules (sICAM-1, sVCAM-1, sE-selectin) and of complement cleavage products (sC4d and sC5b-9), were determined by standardized ELISA in 30 normal controls and 80 samples from 49 recipients of renal allografts. In contrast to the low amounts of adhesion molecules and complement components uniformly excreted by healthy persons (group 0), marked differences were observed among allograft recipients. To prove the clinical relevance of these differences in excretion, patient samples were assigned to 5 categories according to clinical and histopathological criteria: group I--acute steroid-resistant rejection (n = 10); group II--acute steroid-sensitive rejection (n = 10); group III--chronic rejection (n = 23); group IV--stable graft function (n = 27); and group V--miscellaneous disorders (n = 10), including infections, CsA overdoses, and glomerulonephritis. Urinary levels of sICAM-1, sVCAM-1, and sC4d were significantly higher in group I compared with all other groups (P < 0.01). The difference in sICAM-1 excretion between groups III and IV also reached statistical significance (P < 0.05). Urinary concentrations of sICAM-1, sVCAM-1, and sC4d were reflective of their histological distribution in corresponding graft biopsies. None of the patients excreted E-selectin in detectable amounts. Excretion of the terminal membrane attack complex C5b-9 was not significantly associated with any diagnosis. It is concluded that for clinical purposes the combined evaluation of sICAM-1, sVCAM-1, and sC4d is most useful and can provide valuable information with regard to the severity and the type of allograft rejection.
Assuntos
Moléculas de Adesão Celular/urina , Complemento C4/urina , Complemento C4b , Proteínas do Sistema Complemento/urina , Glicoproteínas/urina , Molécula 1 de Adesão Intercelular/urina , Transplante de Rim , Fragmentos de Peptídeos/urina , Complexo de Ataque à Membrana do Sistema Complemento , Selectina E , Rejeição de Enxerto/urina , Humanos , Transplante de Rim/imunologia , Monitorização Imunológica , Solubilidade , Transplante Homólogo , Molécula 1 de Adesão de Célula VascularRESUMO
The urinary excretion of alpha-1-micro-albumin (alpha 1m) and complement components (CCs) was evaluated in the urine of 49 patients suffering from chronic glomerulonephritis (GN). Nephrotic syndrome (NS) was shown in 18 cases and increased serum creatinine level (Pcr: 115.0-159.1 mumol/l) in 9 patients. The most frequent CCs presence and the highest values of alpha 1m excretion were found in patients with membranous GN. In the early phase of the disease the alpha 1m urinary excretion was higher in subjects with NS than in those not showing the feature of it, independently of the morphological basis of the disease. Also CCs were detected mainly in the nephrotic patients. As the glomerular filtration improved a significant decrease in the urinary alpha 1m excretion was observed. The application of steroid immunosuppressive therapy resulted in the decrease of alpha 1m as well as CCs excretion. The results seem to point out that the increased alpha 1m and CCs excretion may be secondary to the activity of glomerular alternations as well as to the disturbances in glomerular blood flow.
Assuntos
alfa-Globulinas/urina , Proteínas do Sistema Complemento/urina , Glomerulonefrite/urina , Síndrome Nefrótica/urina , Adulto , Doença Crônica , Creatinina/sangue , Glomerulonefrite/fisiopatologia , Humanos , Terapia de Imunossupressão , Glomérulos Renais/fisiopatologia , Pessoa de Meia-Idade , Síndrome Nefrótica/fisiopatologiaRESUMO
Deposition of the C5b-9 complex of C in glomeruli of rats with experimental membranous nephropathy (MN) is essential for the development of proteinuria. In this investigation C5b-9 was localized in the passive Heymann nephritis (PHN) by immunoelectron microscopy with a mAb specific for C5b-9(m) neoantigen. Its distribution was compared with that in another model of MN induced by successive injections of cationic human IgG and rabbit anti-human IgG into rats. In PHN C5b-9 was found: 1) in the immune deposits (ID), and on the cell membranes of foot processes close to the ID; 2) in clathrin-coated pits of the glomerular epithelial cells (GEC) close to the ID and in membrane vesicles in the cytoplasm, separated from sheep IgG and the gp330 Ag; 3) in high concentration in multivesicular bodies of GEC; and 4) in association with membrane vesicles in the urinary space which presumably are the exocytosed content of membrane vesicular bodies. By contrast, in the cationic IgG-MN model C5b-9 was found mostly in ID, but rarely within the GEC. By freeze-fracture electron microscopy we have further identified 200- to 250-A intramembrane particles in PHN in the cell membranes of the "soles" of the foot processes which resemble membrane inserted human C5b-9(m). Degradation products of C5b-9 were further detected by immunoblotting of a 100,000 x g pellet of PHN rat urine. These results indicate that, in PHN, C5b-9 is inserted into the cell membranes of GEC, and that it is selectively endocytosed and transported across GEC by a cellular mechanism which apparently protects the cell from accumulation of membrane-inserted C5b-9.
Assuntos
Proteínas do Sistema Complemento/fisiologia , Glomerulonefrite Membranosa/imunologia , Glomérulos Renais/imunologia , Animais , Complexo Antígeno-Anticorpo/análise , Transporte Biológico , Membrana Celular/imunologia , Membrana Celular/patologia , Membrana Celular/ultraestrutura , Complexo de Ataque à Membrana do Sistema Complemento , Proteínas do Sistema Complemento/urina , Endocitose , Epitélio/imunologia , Epitélio/patologia , Epitélio/ultraestrutura , Espaço Extracelular/imunologia , Espaço Extracelular/patologia , Espaço Extracelular/ultraestrutura , Glomerulonefrite/imunologia , Glomerulonefrite/patologia , Glomerulonefrite Membranosa/patologia , Glomerulonefrite Membranosa/urina , Glomérulos Renais/patologia , Glomérulos Renais/ultraestrutura , Masculino , Microcorpos/ultraestrutura , Ratos , Ratos EndogâmicosRESUMO
The third component of complement (C3) was measured in the urine of 98 patients with a variety of renal diseases. Renal biopsy was performed on 83 of the patients and examined by light, electron, and immunofluorescence microscopy. Urinary C3 was detected in cases of membranous glomerulonephritis, mesangiocapillary glomerulonephritis, rapidly progressive glomerulonephritis, and renal amuloidosis. It was not detected in minimal lesion glomerulonephritis; in cases of proliferative glomerulonephritis it was detected only in those showing histological evidence of a progressive lesion. Concentrations were low or undetectable in cases of non-immunological renal diseases. There was a good correlation between urinary C3 concentrations and the deposition of C3 in glomerular capillary walls, as seen by immunofluorescence microscopy, and there was no correlation with the degree or selectivity of proteinuria. Urinary C3 excretion appears to be an accurate indicator of continuing activity of disease. It is suggested that the presence of C3 in urine is due to complement fixation by immune complexes in glomerular capillary walls, and that urinary C3 estimations have potential applications in the study of glomerulonephritis.