RESUMO
BACKGROUND: IgA nephropathy (IgAN) is characterized by deposition in the glomerular mesangium of IgA together with C3, C5b-9, and properdin. IgG deposition as a risk factor in IgAN was recently confirmed by a long-term follow-up of patients with IgAN. We previously reported on an acute model of IgA-mediated glomerular inflammation in Wistar rats. METHODS: To investigate the effect of the combination of IgA and IgG on glomerular injury, Wistar rats were injected with a minimum dose of rat IgG in the presence or absence of a subnephritogenic dose of polymeric rat IgA. Subsequently, glomerular complement activation, influx of inflammatory cells, proteinuria, and hematuria were assessed. RESULTS: Administration of IgG to the rats resulted in maximal proteinuria of 20.3 +/- 12.1 mg/24 h on day 2 and an absence of overt glomerular inflammation. Administration of polymeric rat IgA antibodies to rats resulted in hematuria with a moderate mesangial complement deposition. In the combination group, however, glomerular deposition of C5b-9 was dramatically increased. This was accompanied by increased proteinuria as compared with rats receiving IgA or IgG antibody injections alone on day 7. Microhematuria occurred in rats receiving either polymeric rat IgA or IgG alone or the combination. While both rat IgG and polymeric IgA induced minor mesangial cell (MC) proliferation and MC lysis, the combination resulted in a pronounced, significant increased percentage of aneurysm formation on day 7 after injection. CONCLUSIONS: We conclude that in this model of IgA-induced glomerulopathy, a selective, complement-dependent glomerular inflammation is induced in Wistar rats by glomerular codeposition of rat isotypic monoclonal antibodies.
Assuntos
Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Imunoglobulina A/farmacologia , Imunoglobulina G/farmacologia , Animais , Anticorpos Monoclonais , Complemento C3/análise , Complemento C3/imunologia , Complexo de Ataque à Membrana do Sistema Complemento/análise , Complexo de Ataque à Membrana do Sistema Complemento/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Mesângio Glomerular/imunologia , Mesângio Glomerular/patologia , Hematúria/imunologia , Hematúria/patologia , Imunoglobulina A/análise , Imunoglobulina G/análise , Masculino , Proteinúria/imunologia , Proteinúria/patologia , Ratos , Ratos WistarRESUMO
BACKGROUND: IgA nephropathy (IgAN) is the most common type of immunologically mediated glomerulonephritis (GN) and is characterized by deposition in the glomerular mesangium of IgA together with C3, C5b-9, and properdin. In patients, the codeposition of IgA together with IgG and/or IgM can lead to a more progressive course of disease. In Wistar rats, mesangial proliferative GN can be induced by the injection of mouse IgG anti-Thy-1 antibodies (ER4G). In contrast, the administration of mouse IgA anti-Thy-1 antibodies (ER4A) to rats results in isolated hematuria without detectable albuminuria and without detectable complement deposition. METHODS: To investigate the effect of the combination of IgA and IgG on glomerular injury, Wistar rats were injected with a limiting dose of ER4G in the presence or absence of ER4A in a dose able to induce hematuria. RESULTS: Although the limiting dose of ER4G or the dose of ER4A used did not induce significant albuminuria, the combination of ER4G and ER4A resulted in a synergistic increase in albuminuria. Microhematuria occurred in rats receiving either ER4A or ER4G alone or in combination. Although both ER4A or a limiting dose of ER4G induced minor increases in extracellular matrix expansion, the combination resulted in a pronounced, additive increased matrix expansion. CONCLUSION: We conclude that in this model of IgA-mediated glomerulopathy, a selective complement-dependent synergistic renal injury is induced in Wistar rats by glomerular codeposition of mouse anti-Thy-1 monoclonal isotypes.
Assuntos
Glomerulonefrite por IGA/etiologia , Imunoglobulina A/administração & dosagem , Imunoglobulina G/administração & dosagem , Antígenos Thy-1 , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/metabolismo , Modelos Animais de Doenças , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Humanos , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Rim/imunologia , Rim/patologia , Masculino , Camundongos , Microscopia de Fluorescência , Ratos , Ratos WistarRESUMO
Mesangial cell (MC) injury is a characteristic feature in the early phase of Thy.1 nephritis. The present study investigates the contribution of complement to MC apoptosis in this experimental model of kidney disease in rats. Thy.1 nephritis was induced by injection of mouse anti-Thy.1 monoclonal antibody (ER4G). To assess the contribution of the terminal sequence of complement on apoptosis, the studies were performed in complement-sufficient PVG/c (PVG/c+) rats and in rats deficient in complement C6 (PVG/c-). Apoptosis was monitored by assessment of the number of condensed nuclei in kidney sections stained with periodic acid-Schiff (PAS) and by the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method and expressed as number of apoptotic cells per 50 glomerular cross sections. In the PAS method, 1 h after intravenous injection of ER4G, PVG/c+ rats exhibited 160.9 +/- 49.5 apoptotic cells, whereas PVG/c- rats had only 3.2 +/- 1.4 apoptotic cells. Control rats exhibited 0.9 +/- 0.6 apoptotic cells. These findings were confirmed with the TUNEL method. In PVG/c- rats, a maximum number of 8.8 +/- 3.1 TUNEL-positive (TUNEL+) cells was found at 6 h followed by a decline thereafter. In PVG/c+ rats, apoptosis was associated with deposition of C6 and C5b-9. Restoration of the complement system of PVG/c- rats with purified human C6 resulted in an increase of apoptosis at 1 h after injection of ER4G from minimal numbers to 239.9 +/- 52.4 TUNEL+ cells. These studies appear to indicate for the first time that the terminal sequence of complement is involved in induction of apoptosis.
Assuntos
Apoptose/imunologia , Complexo de Ataque à Membrana do Sistema Complemento/imunologia , Mesângio Glomerular/imunologia , Mesângio Glomerular/ultraestrutura , Nefrite/imunologia , Nefrite/patologia , Análise de Variância , Animais , Anticorpos Monoclonais/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular , Células Cultivadas , Cromatina/ultraestrutura , Complexo de Ataque à Membrana do Sistema Complemento/genética , Relação Dose-Resposta a Droga , Masculino , Camundongos , Microscopia de Fluorescência , Ratos , Ratos Endogâmicos , Ratos Sprague-Dawley , Valores de Referência , Especificidade da Espécie , Antígenos Thy-1/imunologiaRESUMO
An important antiinflammatory mechanism of intravenous immunoglobulin preparations (IVIG) is their ability to block complement activation. The purpose of this study was to compare the complement-inhibitory activity of four IVIG preparations differing in isotype composition. The preparations were: (1) IVIgG (48 g/L IgG, 2 g/L IgA; Intraglobin F); (2) Pentaglobin (38 g/L IgG, 6 g/L IgM, 6 g/L IgA); (3) IVIgM (35 g/L IgM, 12 g/L IgA, 3 g/L IgG); and (4) IVIgA (41 g/L IgA, 9 g/L IgG), all from Biotest Pharma GmbH, Dreieich, Germany. Their complement inhibitory activity was assessed in vitro by measurement of the blocking of C1q-, C4-, and C3 deposition on solid-phase aggregated rabbit IgG by enzyme-linked immunosorbent assay (ELISA). Complement inhibition in this ELISA was best for IVIgM, followed by Pentaglobin and IVIgG; IVIgA did not exhibit an inhibitory effect. Control experiments with excess concentrations of C1q as well as with C1q-depleted serum showed that the inhibitory effects of IVIG were not caused by complement activation and thus, consumption, but that C4 and C3 were scavenged by IgM and to a lesser extent by IgG. These results were confirmed in vivo in the rat anti-Thy 1 nephritis model, in which a single dose of 500 mg/kg of IVIgM prevented C3-, C6-, and C5b-9 deposition in the rat glomeruli, whereas the effect of IVIgG was much less pronounced. Reduction of complement deposition was paralleled by a diminished albuminuria, which was completely absent in the IVIgM-treated rats. IVIgM and to a lesser extent IVIgG also prevented rat C3 deposition on cultured rat glomerular mesangial cells in vitro, but did not influence anti-Thy 1 binding. Neither IVIgM nor Pentaglobin nor IVIgG negatively affected in vitro phagocytosis of Escherichia coli (E coli) by human granulocytes. In conclusion, we have shown that IgM enrichment of IVIG preparations enhances their effect to prevent the inflammatory effects of complement activation.
Assuntos
Ativação do Complemento/efeitos dos fármacos , Imunoglobulina M/farmacologia , Imunoglobulinas Intravenosas/farmacologia , Inflamação/terapia , Animais , Anticorpos Monoclonais/toxicidade , Complemento C3a/análise , Complemento C4/metabolismo , Proteínas do Sistema Complemento/análise , Depressão Química , Mesângio Glomerular/imunologia , Humanos , Imunoglobulina G/metabolismo , Imunoglobulina M/metabolismo , Imunoglobulina M/uso terapêutico , Imunoglobulinas Intravenosas/uso terapêutico , Inflamação/imunologia , Masculino , Nefrite/etiologia , Nefrite/imunologia , Nefrite/terapia , Fagocitose/efeitos dos fármacos , Coelhos , Ratos , Ratos Wistar , Antígenos Thy-1/imunologiaRESUMO
OBJECTIVE: Forearm blood flow plethysmography is a widely accepted in vivo technique for pharmacologic and functional studies in peripheral resistance vessels and veins. Pharmacological effects on forearm blood flow (FBF) are usually expressed by means of dose-response relationships. This approach does not consider the influence of variations in FBF on the actual plasma concentrations of compounds infused, and is less suitable for quantitative comparison of the pharmacologic characteristics of different compounds. The aim of this study was to validate an equation to estimate the plasma concentrations of intra-arterially infused compounds. This was done at different levels of FBF, using an indicator dilution technique with constant rate infusions of indocyanine green (ICG) and inulin. METHODS: ICG (0.5 mg/min) and insulin (5 mg/min) were infused into the brachial artery in the presence of sodium nitroprusside (10 ng/kg/min; to obtain high FBF), vehicle (0.9% saline; for intermediate FBF), and methoxamine (1 microgram/kg/min; for low FBF), FBF was measured using venous occlusion plethysmography in six healthy male volunteers. Plasma concentrations of the indicators, measured in venous blood samples, were compared with the calculated values. RESULTS: Excellent correspondence was observed between calculated and measured plasma concentrations for both ICG and inulin. Venous plasma concentrations of ICG (> or = 95% protein binding) reached steady-state within four min independent of FBF. Alternatively, the time required for venous plasma concentrations of inulin (not bound to protein) to reach steady-state appeared dependent on FBF. CONCLUSION: Total plasma concentrations of intra-arterially infused drugs can be appropriately estimated at the level of the arterioles by the proposed equation.
Assuntos
Antebraço/irrigação sanguínea , Verde de Indocianina/análise , Verde de Indocianina/farmacologia , Inulina/sangue , Inulina/farmacologia , Adulto , Humanos , Infusões Intra-Arteriais , Masculino , Metoxamina/farmacologia , Nitroprussiato/farmacologia , Pletismografia , Fluxo Sanguíneo Regional/efeitos dos fármacos , Estatísticas não Paramétricas , Vasoconstritores/farmacologia , Vasodilatadores/farmacologiaRESUMO
A PVG rat with total deficiency of C6 and partial deficiency of C2 (PVG/c-), and a syngeneic control strain (PVG/c+), were used to study the production of extrahepatically synthesized complement. Livers of complement deficient rats were transplanted in sufficient rats (Tx-L). The C6 and C2 levels in Tx-L rats declined within 2 days to 25% and 30%, respectively, and remained stable for more than 6 weeks. To investigate the contribution of C6 synthesis by the liver, C6 sufficient livers were grafted in deficient rats (Tx + 1). After an initial increase, with maximum C6 levels of 119% at 10 days following transplantation, the C6 levels decreased gradually and C6 was no longer detectable 28 days after transplantation. This decline in C6 levels was dependent on antibody production against C6. No significant change in the C3, C4, factor H and factor B levels was observed. Expression of C6 mRNA in the grafted PVG/c+ sufficient liver was comparable to the expression of C6 mRNA in control PVG/c+ livers while C6 mRNA expression in the transplanted PVG/ c- liver and the control PVG/c- liver was lower. In conclusion, it was demonstrated in vivo that not only C6 but also C2 is synthesized extrahepatically in PVG/c rats.
Assuntos
Complemento C2/biossíntese , Complemento C2/genética , Complemento C6/biossíntese , Complemento C6/genética , Ratos Endogâmicos/imunologia , Animais , Anticorpos/sangue , Complemento C2/deficiência , Complemento C6/deficiência , Complemento C6/imunologia , Ensaio de Atividade Hemolítica de Complemento , Fígado/imunologia , Fígado/metabolismo , Transplante de Fígado/imunologia , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos/genética , Transplante IsogênicoRESUMO
Complement C6 plays an important role in the effector phase of complement-mediated cell lysis. Recently, a PVG/c rat strain deficient in haemolytic C6 activity was discovered. In the present study we show that these rats lack both antigenic and functional C6, and that repetitive immunization of these rats with PVG/c+ serum results in generation of specific anti-rat C6 antibodies. The observed absence of rat C6 was further investigated at the genomic and transcriptional level using a 492-bp cDNA of rat C6, cloned from a rat liver cDNA library using full length human C6 as a probe. Northern blot analysis revealed the presence of C6 mRNA in livers of both PVG/c- and PVG/c+ rats, corresponding to a size of approximately 3.3 kb, although the level of C6 mRNA expression was approximately 100-fold less in PVG/c- rats. In addition, using rat C6-specific primers, positive signals were obtained in kidneys of both rat strains by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Southern blot analysis of digested genomic DNA did not reveal evidence for large C6 gene deletions. We conclude that the lack of C6 protein in the PVG/c- rat strain is not due to a (large) C6 gene deletion, but presumably is caused by an unstable mRNA or a point mutation in the C6 gene resulting in an aberrant transcription of the C6 gene. Alternatively, a gene coding for a product involved in C6 biosynthesis that acts in trans may carry a mutation.
Assuntos
Complemento C6/deficiência , Complemento C6/genética , Glomerulonefrite Membranoproliferativa/genética , Ratos Mutantes/genética , Animais , Sequência de Bases , Cromatografia de Afinidade , Cromatografia em Gel , Ativação do Complemento/imunologia , Complemento C6/imunologia , Ensaio de Atividade Hemolítica de Complemento , DNA/análise , Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranoproliferativa/patologia , Imunização , Immunoblotting , Rim/patologia , Dados de Sequência Molecular , Mutação Puntual/genética , RNA Mensageiro/genética , Ratos , Ratos Mutantes/imunologiaRESUMO
In order to study the contribution of extrahepatic C6 to anti-Thy1.1 nephritis, C6 deficient PVG/c- livers were grafted in C6 sufficient PVG/c+ rats (Tx-L). Infusion of anti-Thy1.1 antibodies in Tx-L and PVG/c+ rats resulted in generation of C5b-9 complexes and subsequent glomerular injury, while infusion of anti-Thy1.1 antibodies in PVG/c- rats revealed no detectable C6 deposition. Because C6 mRNA was expressed in both liver and kidney tissue of PVG/c+ rats, we assessed whether production of C6 in the kidney alone was sufficient for glomerular injury. One kidney of a PVG/c- rat was replaced with a PVG/c+ kidney (Tx + K) followed by administration of anti-Thy1.1 antibodies. C6 deposits were detectable neither in PVG/c+ kidneys nor in PVG/c- kidneys of Tx + K rats, indicating that C6 production in PVG/c+ kidneys alone is not sufficient to contribute to renal injury. That C6 production had occurred was suggested by the presence of equal amounts C6 mRNA in control PVG/c+ kidneys and in grafted PVG/c+ kidneys of Tx + K rats. C6 mRNA expression in kidney tissue of PVG/c+ rats is presumably derived from peritubular sites. In conclusion, we have demonstrated that extrahepatic, but not renal synthesis of, C6 is sufficient to contribute to glomerular injury during anti-Thy1.1 nephritis.
Assuntos
Complemento C6/metabolismo , Complexo de Ataque à Membrana do Sistema Complemento/metabolismo , Rim/metabolismo , Fígado/metabolismo , Animais , Biomarcadores , Complemento C2/deficiência , Complemento C6/deficiência , Complemento C6/genética , Imunofluorescência , Transplante de Rim , Transplante de Fígado , Nefrite/imunologia , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos/sangue , Ovinos/sangue , Antígenos Thy-1/imunologiaRESUMO
The interactions of intrarenal dopamine (DA) synthesis and natriuresis were studied in 11 healthy volunteers by giving a low and high sodium diet (LoSo, 50 mmol Na+; HiSo, 250 mmol Na+). On 2 days in both dietary phases an acute saline load was given, without or with an infusion of the DA precursor 3, 4-dihydroxyphenylalanine (DOPA) 0.15 microgram kg-1 min-1. Urinary excretion rates of sodium (UNa V), DOPA (UDOPA V), DA (UDA V) and noradrenaline (NA; UNA V), hormonal parameters, blood pressure (BP), glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were measured. On HiSo UDOPA V increased by 35% (P < 0.05) while UDA V remained unchanged. Only weak correlations were found between 24h UNa V and UDA V (r = 0.23) or UDOPA V (r = 0.39). On both diets and without the DOPA infusions, the saline infusions enhanced UNa V but did not increase UDA V. The DOPA infusions caused no significant change of UNa V, in spite of five- to eight-fold increases in UDA V. During LoSo the DOPA infusion only caused a slightly larger increase of UNa V after the saline infusion, when expressed as percentage change to the pre-saline UNa V (119 +/- 25% without DOPA infusion, 244 +/- 56%, P < 0.05 with DOPA infusion). On HiSo the DOPA infusion did not affect UNA V. The dietary sodium intake and the DOPA infusions did not influenced BP, GFR, or ERPF. In conclusion, dietary sodium intake or acute sodium loads did not clearly modulate UDA V.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Di-Hidroxifenilalanina/administração & dosagem , Dopamina/urina , Rim/metabolismo , Sódio na Dieta/administração & dosagem , Sódio/urina , Adulto , Pressão Sanguínea/efeitos dos fármacos , Taxa de Filtração Glomerular/efeitos dos fármacos , Humanos , Rim/fisiopatologia , Masculino , Natriurese , Fluxo Plasmático Renal/efeitos dos fármacos , Sódio/administração & dosagemRESUMO
BACKGROUND: In autosomal dominant polycystic kidney disease (ADPKD) the pathophysiology of hypertension, which is frequently observed before loss of renal function, is not well understood. We investigated intrarenal dopamine, the renin-angiotensin-aldosterone system (RAAS), and plasma endothelin in relation to sodium homeostasis as potential hypertensive factors in this disease. METHODS: Eight borderline hypertensive ADPKD patients with (near) normal renal function and seven matched healthy control subjects were investigated at three levels of daily dietary sodium intake: 150, 50 and 450 mmol. In the 450-mmol sodium intake period we studied the effects of renally formed dopamine by infusing its precursor DOPA (DOPAi.v., 7 micrograms kg-1 min-1). In the 50-mmol sodium intake period we studied the influence of the RAAS by administering enalaprilate (42 micrograms kg-1), followed by angiotensin II (12 ng kg-1 min-1) intravenously. GFR and ERPF were measured by continuous infusion of inulin and PAH. RESULTS: At all levels of sodium intake sodium balances were equal, but daily urinary excretions of dopamine and DOPA were higher (P < 0.01) in the ADPKD patients than in the controls. Renal vascular resistance, filtration fraction and blood pressure were higher in the ADPKD patients (all P < 0.05) while plasma renin activity was similar. DOPAi.v. normalized renal haemodynamics and increased plasma endothelin in ADPKD patients (all P < 0.05), while stimulation of natriuresis was equal in both groups. Enalaprilate increased plasma endothelin in the ADPKD patients and only partially normalized renal haemodynamics. CONCLUSIONS: In borderline hypertensive ADPKD patients: (1) urinary dopamine excretion is increased at all levels of sodium intake, suggesting that this may be needed to maintain sodium balance; (2) stimulation of renal dopamine production is able to normalize renal haemodynamics, making dopamine receptor agonism a potential therapeutic option; (3) the activity of the RAAS is not clearly enhanced; (4) renal vasodilatation increases plasma endothelin levels.
Assuntos
Di-Hidroxifenilalanina/uso terapêutico , Dopaminérgicos/uso terapêutico , Dopamina/metabolismo , Hipertensão Renal/metabolismo , Rim Policístico Autossômico Dominante/metabolismo , Adulto , Aldosterona/sangue , Angiotensina II/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Enalaprilato/administração & dosagem , Endotelinas/sangue , Endotelinas/efeitos dos fármacos , Feminino , Hemodinâmica/efeitos dos fármacos , Humanos , Hipertensão Renal/tratamento farmacológico , Hipertensão Renal/etiologia , Infusões Intravenosas , Masculino , Pessoa de Meia-Idade , Rim Policístico Autossômico Dominante/complicações , Rim Policístico Autossômico Dominante/fisiopatologia , Sódio/urina , Sódio na Dieta/administração & dosagemRESUMO
The presence of anti-glomerular basement membrane antibodies is one of the features of Goodpasture's syndrome. Since the disease has a rapidly progressive course, an early diagnosis is essential. As was already demonstrated in other ELISA methods, 2.45-GHz microwave irradiation can accelerate all kinds of time consuming processes in several laboratory techniques. The application of microwaves in an ELISA for the measurement of anti-GBM antibodies in serum indicated that a considerable time reduction of 75% can be achieved, resulting in a rapid and reliable assay. In addition, microwaves can also have a positive effect on the resolution of that particular ELISA as shown in this study.
Assuntos
Doença Antimembrana Basal Glomerular/imunologia , Autoanticorpos/análise , Membrana Basal/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Glomérulos Renais/imunologia , Humanos , Micro-Ondas , Reprodutibilidade dos TestesRESUMO
The prevalence of antibodies against the collagen-like region of the subcomponent of the first component of complement, C1q, was investigated in 11 patients with anti-glomerular basement membrane (GBM) nephritis. Anti-C1q antibodies (anti-C1qAb) were detected in seven patients. IgG anti-C1qAb were found in four and IgA anti-C1qAb in five patients. During follow up of the patients a relationship was observed between the levels of IgG anti-C1qAb and the levels of anti-GBM antibodies (anti-GBMAb). Gelfiltration experiments indicated that both IgG anti-C1qAb as well as IgG anti-GBMAb were monomeric and that binding also occurred with the F(ab')2 fragments of the antibodies. Although anti-C1qAb and anti-GBMAb are both directed against a collagen-like structure, it was demonstrated by means of inhibition experiments that anti-C1qAb and anti-GBMAb are directed against different antigenic sites. Comparison of patients with anti-GBM nephritis with and without anti-C1qAb revealed that there were no differences in disease activity or disease severity. Therefore, the results of this study suggest that anti-C1qAb do not play a direct pathogenetic role in anti-GBM nephritis.
Assuntos
Autoanticorpos/imunologia , Membrana Basal/imunologia , Complemento C1q/imunologia , Glomerulonefrite/imunologia , Adulto , Idoso , Humanos , Glomérulos Renais/imunologia , Pessoa de Meia-IdadeRESUMO
In the present study we treated rats with N-nitrosodimethylamine (DMN) or D-galactosamine (GALN) to achieve increased circulating IgA levels in rats. GALN-treated rats showed a six-fold increase in serum IgA levels after the first intraperitoneal (i.p.) injection, whereas a 10-fold increase after a second i.p. injection of GALN was seen. DMN-treated rats showed a three-fold increase in serum IgA levels. No differences were observed in IgG and IgM levels between treated and non-treated rats. Sequential renal biopsies analysed by immunofluroescence exhibited mesangial deposits of IgA with different intensities of C3 deposition. Rats treated with GALN showed more IgA deposition in the kidney than DMN-treated rats. The IgA deposition together with C3 was more prominent in rats treated with GALN than in rats treated with DMN. The deposition of C3 together with IgA was associated with an influx of monocytes as detected by ED-1, an antibody directed against a rat monocyte marker. These studies provide evidence that an increase in serum IgA levels is associated with deposition of IgA in the kidney and that IgA has an inflammatory potential.
Assuntos
Glomerulonefrite por IGA/imunologia , Imunoglobulina A/análise , Macrófagos/imunologia , Animais , Complemento C3/análise , Dimetilnitrosamina/administração & dosagem , Modelos Animais de Doenças , Imunofluorescência , Galactosamina/administração & dosagem , Glomerulonefrite por IGA/sangue , Glomerulonefrite por IGA/patologia , Imunoglobulina G/análise , Imunoglobulina M/análise , Masculino , Ratos , Ratos EndogâmicosRESUMO
The immunogenicity of the minor histocompatibility antigen Eag-1 of the rat kidney endothelium was studied in renal allografts mismatched for antigens of the major histocompatibility complex (MHC). The rejection rates of BN.1L (RT1(1), Eag-1+), (BN.1LXMAXX)F1 (RT1l/n, Eag-1+), and LEW (RT1l, Eag-1-) kidneys transplanted into unsensitized, bilaterally nephrectomized MAXX (RT1n, Eag-1-) recipients were comparable, indicating that incompatibility for Eag-1 has no effect on the survival of MHC-incompatible kidney grafts. Transplantation of BN (RT1n, Eag-1+) and WKY (RT1k, Eag-1+) kidneys into unilaterally nephrectomized MAXX recipients led to a weak and inconsistent antibody response against Eag-1, indicating that MHC incompatibility does not influence the formation of antibodies against Eag-1.
Assuntos
Endotélio/imunologia , Rejeição de Enxerto , Isoantígenos/imunologia , Transplante de Rim , Animais , Sítios de Ligação de Anticorpos , Doença Enxerto-Hospedeiro/imunologia , Doença Enxerto-Hospedeiro/patologia , Antígenos de Histocompatibilidade/análise , Antígenos de Histocompatibilidade/imunologia , Teste de Histocompatibilidade , Isoanticorpos/biossíntese , Rim/metabolismo , Rim/patologia , Masculino , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos Lew , Ratos Endogâmicos WKYRESUMO
Immunizations of MAXX rats with spleen and lymph node cells from BN donors lead to the production of antibodies against the renal endothelial antigen Eag-1. In the present study we analyzed the ability of various tissues to induce a response against Eag-1. Lung homogenates, like spleen and lymph node cells, induced a strong and rapid antibody response. A weak and slow response was induced by cardiac allografts as well as kidneys transplanted into recipients in which unilateral or delayed bilateral nephrectomy was performed. Transplantation of kidney or skin allografts into intact recipients did not result in the formation of Eag-1 antibodies, nor did immunizations with kidney homogenates. A rapid response, however, was observed after transplantation of BN kidneys into MAXX rats previously sensitized by a single injection of BN lymphoid cells, whereas no secondary response was induced by BN kidney homogenates. With an indirect immunofluorescence technique, Eag-1 was detected on lung and kidney endothelium, but not on lymphoid cells or heart and skin tissue. It thus appears that the immunogenicity of Eag-1 does not correlate with the quantitative expression in different tissues and that lymphoid cells, rather than vascular endothelial cells, can induce a strong and rapid response against Eag-1. Because the immunogenicity of lymphoid cells was diminished by heat treatment and freeze-thaw lysis, it appears that intact donor cells are required for the stimulation of antibody production.
Assuntos
Antígenos/imunologia , Endotélio/imunologia , Complexo Principal de Histocompatibilidade , Animais , Formação de Anticorpos , Células Apresentadoras de Antígenos/fisiologia , Congelamento , Transplante de Coração , Temperatura Alta , Imunização , Transplante de Rim , Pulmão/citologia , Tecido Linfoide/citologia , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos , Circulação Renal , Transplante de Pele , Imunologia de Transplantes , Transplante Homólogo , Irradiação Corporal TotalAssuntos
Antígenos de Superfície/análise , Biomarcadores/análise , Sobrevivência de Enxerto/imunologia , Teste de Histocompatibilidade , Transplante de Rim/imunologia , Glicoproteínas de Membrana/análise , Animais , Antígenos de Superfície/imunologia , Imunofluorescência , Rim/imunologia , Glicoproteínas de Membrana/imunologia , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos LewRESUMO
Previous studies have shown that immunization of MAXX rats with spleen and lymph node cells from the MHC-identical BN strain results in the formation of antibodies that react with the renal endothelial alloantigen Eag-1. In the present study, the reactivity of MAXX anti-BN sera was further characterized. No reactivity of the antisera was detected with unseparated spleen, lymph node, thymus and bone marrow cell suspensions, peripheral blood, or cells obtained from lung lavages. The antisera did, however, react with splenic macrophages, as well as with peritoneal granulocytes and macrophages from BN, BN.1A, BN.1L, and PVG rats. Genetic studies revealed that the antigen, provisionally designated Pag-1, segregates independently of Eag-1, the RT1 complex, sex, and the hooded and albino traits. Pag-1 appears to be absent in the kidney, since absorption of MAXX anti-BN sera with BN kidney homogenates did not remove the reactivity against Pag-1, and antisera raised against BN peritoneal cells did not bind with the renal endothelium. Pag-1 is expressed on bone marrow-derived cells, since peritoneal cells from lethally irradiated MAXX rats that were reconstituted with bone marrow cells from BN donors reacted with MAXX anti-BN sera, whereas peritoneal cells from BN rats reconstituted with MAXX bone marrow did not.