RESUMEN
The nephrotic syndrome is a rare complication of allogeneic stem cell transplantation (alloHSCT). We present two cases of nephrotic syndrome during chronic graft-versus-host disease (GvHD) involving altered cytokine gene expression in renal tissue. A patient with acute lymphatic leukemia demonstrated nephrotic syndrome due to minimal change disease as a marker of chronic GvHD. A patient with acute lymphoblastic leukemia suffered from severe nephrotic syndrome due to membranous glomerulopathy. In the two presented cases of GvHD-linked nephrotic syndrome, increased cytokine gene expression [tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), interferon gamma (IFN-gamma), interleukin 2 (IL-2), IL-6, and IL-10] assessed using semiquantitative evaluation with reverse transcriptase polymerase chain reaction (RT-PCR) in situ on renal biopsy was observed.
Asunto(s)
Citocinas/genética , Enfermedad Injerto contra Huésped/complicaciones , Nefrosis Lipoidea/etiología , Síndrome Nefrótico/etiología , Adulto , Citocinas/metabolismo , Diagnóstico Diferencial , Expresión Génica , Enfermedad Injerto contra Huésped/genética , Humanos , Masculino , Nefrosis Lipoidea/diagnóstico , Nefrosis Lipoidea/genética , Síndrome Nefrótico/diagnóstico , Síndrome Nefrótico/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
UNLABELLED: The aim of the study was to evaluate the effect of brain death, ischemia-reperfusion injury and alloreactivity of some cytokines on intragraft mRNA expression. MATERIAL AND METHODS: We have examined 49 needle core biopsies of kidney allografts from cadaveric donors. Samples were taken before harvesting, after cold ischemia and approximately after 20-30 minutes of reperfusion. We have also assessed 56 allograft biopsies taken after transplantation. Tubular and glomerular expression of IL-2, IL-6, IL-10, IFN-gamma, TGF-beta 1 and PDGF-B mRNA was assessed using semiquantitative evaluation of the RT-PCR in situ on paraffin tissue sections. RESULTS: In all pre-procurement specimens high glomerular and tubular IL-2, IL-6, IL-10, TGF-beta 1, PDGF-B and IFN-gamma mRNA expression was detected. After reperfusion an increase of IL-2, IL-6, and IL-10 mRNA expression was observed in all specimens and was limited only to tubules. Biopsies samples with borderline changes exhibited the lowest levels of cytokine gene expression close to the intensity in control specimens. An intense, comparing to normal kidney, tubular and glomerular all examined cytokines gene expression was also noticed during acute rejection. No significant differences between acute cellular and vascular rejection were noticed. The mRNA expression of IFN-gamma and IL-2, IL-6, IL-10 in chronic rejection did not differ from acute rejection. The tubular expressions of mRNA for IL-6 and TGF-gamma 1 in biopsies with acute rejection obtained from patients treated with MMF were significantly lower than in biopsies obtained from patients treated with azathioprine.
Asunto(s)
Expresión Génica/genética , Interleucinas/genética , Trasplante de Riñón/patología , Riñón/fisiopatología , Factor de Crecimiento Derivado de Plaquetas/genética , Factor de Crecimiento Transformador beta/genética , Adolescente , Adulto , Anciano , Biopsia con Aguja , Cadáver , Femenino , Humanos , Riñón/patología , Trasplante de Riñón/métodos , Donadores Vivos , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
The aim of our study was to correlate intragraft mRNA expression of cytokines and growth factors with histopathologic features in renal allograft biopsies. Fifty-six core biopsies performed in 51 kidney transplant recipients were assessed by the Banff '97 classification. Tubular and glomerular expressions of IFN-gamma, TGF-beta1, and PDGF-B as well as IL-2, IL-6, and IL-10 mRNA were assessed using semiquantitative RT-PCR in situ. No significant differences were noted between acute cellular and vascular rejection with regard to the glomerular and tubular mRNA expression of cytokines examined. We observed a positive correlation between tubular and glomerular IL-10 and IFN-gamma mRNAs during acute rejection. In chronic rejection the mRNA expression levels of IFN-gamma and IL-2, IL-6, and IL-10 did not differ from those of acute rejection; moreover, the glomerular expression of mRNA for TGF-beta1 (P < .05) and PDGF-B (P < .1) was even lower than during acute rejection episodes. Both tubular and glomerular IL-2, TGF-beta1, and PDGF-B mRNA expression levels in biopsies with acute rejection were significantly higher than in acute tubular necrosis (ATN). Biopsy samples with borderline changes exhibited the lowest levels of cytokine gene expression and were close to the intensity of control specimens obtained from living donor kidney biopsies taken during organ harvest. Our data failed to show a dichotomy between Th1 and Th2 cytokine activation in biopsy specimens from kidney allograft recipients; both Th1- and Th2-derived cytokines were involved to similar extents in rejection processes.
Asunto(s)
Citocinas/genética , Sustancias de Crecimiento/genética , Trasplante de Riñón/inmunología , ARN Mensajero/genética , Biopsia , Humanos , Interleucinas/genética , Glomérulos Renales/inmunología , Glomérulos Renales/patología , Trasplante de Riñón/patología , Túbulos Renales/inmunología , Túbulos Renales/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Transcripción GenéticaRESUMEN
The aim of the study was to assess the molecular background of the alloimmune response by the detection of low-abundance mRNA of cytokines in 34 core needle biopsies from kidney allografts with histopathological findings of acute rejection (AR). Recipients were immunosuppressed with a calcineurin inhibitor (CNI), cyclosporine or tacrolimus, and prednisone and azathioprine or mycophenolate mofetil (MMF). Tubular and glomerular expression of IL-2, IL-6, IL-10, IFN-gamma, TGF-beta1, and PDGF-B mRNA were assessed using semiquantitative evaluations of RT-PCR in situ on paraffin tissue sections. This procedure resulted in light microscopy visualization of granular precipitates at the sites of the corresponding mRNA chains. The tubular expression of mRNA for IL-6 and TGF-beta1 was significantly lower in biopsies with AR (n = 34) obtained from patients treated with MMF (n = 12) than in biopsies obtained from patients treated with azathioprine (n = 22) (P < .02). Responsiveness to corticosteroids tended to be more frequent among the MMF group (11 of 12 recipients vs 15 of 22 recipients, P = ns). Moreover, 8 of 12 recipients in the MMF-treated group displayed serum creatinine levels equal or less than 167 mmol/L 1 year after biopsy compared to 7 of 22 recipients in the azathioprine-treated group. There was no significant difference between the groups that had or had not received corticosteroids or between those treated with each type of CNI. These results suggest stronger inhibition of humoral responses and down-regulation of fibrosis by MMF among recipients with AR.
Asunto(s)
Ciclosporina/uso terapéutico , Citocinas/genética , Trasplante de Riñón/inmunología , Ácido Micofenólico/análogos & derivados , Tacrolimus/uso terapéutico , Transcripción Genética/inmunología , Biopsia , Inhibidores de la Calcineurina , Quimioterapia Combinada , Sustancias de Crecimiento/genética , Humanos , Inmunosupresores/uso terapéutico , Trasplante de Riñón/patología , Túbulos Renales/inmunología , Túbulos Renales/patología , Donadores Vivos , Ácido Micofenólico/uso terapéutico , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Trasplante Homólogo/inmunología , Trasplante Homólogo/patologíaRESUMEN
It was previously reported that ischemia-reperfusion injury initiates an inflammatory response and may significantly affect the transplanted organ function. The aim of this study was to assess changes of intragraft cytokine mRNA expression in kidneys after cold ischemia (CI) and following reperfusion. We examined mRNA of a product of activated T lymphocytes (IFN-gamma) and a monocyte product (IL-6). Eleven kidneys were transplanted after CI time ranging from 16 to 39 hours. Renal needle core biopsies were obtained from donors after cold ischemia and approximately after 20 minutes of reperfusion. Tubular and glomerular expression of IFN-gamma and IL-6 mRNA were assessed using semiquantitative evaluation of the RT-PCR in situ. After reperfusion an intense increase of IL-6 mRNA expression was observed in four specimens, a slight increase was noticed in five specimens, and a very slight decrease in two specimens. Changes in IL-6 mRNA expression were limited only to tubules. In contrast, the glomerular and tubular mRNA expression of IFN-gamma and glomerular of IL-6 remained stable. Mean CI time for patients with an intense increase was higher than for patients with a slight increase and with the decrease of IL-6 mRNA expression (32.0 +/- 6.8 vs 25.2 +/- 7.3 and 26.0 +/- 5.7 hours). Our results suggest that early inflammatory changes at the time of implantation of renal allografts depends mainly on monocyte/macrophage-associated products. The observed intensity of their expression in tubules was connected to longer CI time.
Asunto(s)
Citocinas/genética , Trasplante de Riñón/inmunología , Adolescente , Adulto , Anciano , Biopsia con Aguja , Femenino , Humanos , Interleucina-6/genética , Isquemia , Riñón , Trasplante de Riñón/métodos , Trasplante de Riñón/patología , Masculino , Persona de Mediana Edad , Preservación de Órganos/métodos , ARN Mensajero/genética , Reperfusión , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
The reductive methylation procedure for proteins in solution has been adapted for tritium radiolabelling of surface components of macrophages, including the Fc gamma receptor. High specific radioactivity of the receptor was obtained with no loss of IgG binding activity, tested by the solid-phase radioassay. To prepare the support for the assay, cellulose filter paper was CNBr-activated and a diamine hydrocarbon reagent was attached as the spacer chain. Next, TNP-hapten residues were covalently coupled to free amine groups. Antibody-coated TNP-paper discs were used as the solid phase for the competitive binding radioassay for the determination of IgG-binding activity of purified Fc gamma receptor. The developed radioassay permits the detection of low affinity interactions and/or low quantities of Fc receptors.
Asunto(s)
Macrófagos/ultraestructura , Radioinmunoensayo/métodos , Receptores Fc/metabolismo , Animales , Cobayas , Inmunoglobulina G , Papel , Cavidad Peritoneal/citología , Receptores Fc/antagonistas & inhibidores , Receptores Fc/aislamiento & purificación , Receptores de IgG , Solubilidad , Trinitrobencenos , TritioRESUMEN
Comparative studies of physiocochemical properties of ovine and bovine serum and colostral IgM are described. Serologic properties, molecular weight, amino acid composition, carbohydrate content, peptide maps, heterogeneity, and ORD and CD spectra were compared. Colostral IgM have additional antigenic determinants, higher carbohydrate content, lower heterogeneity, and higher content of beta-structure. Different amino acid composition and different peptide maps of serum and colostral IgM were shown. The results obtained suggest that ovine and bovine colostral IgM immunoglobulins may be a mixed population of molecules locally synthesized and transferred from the serum.
Asunto(s)
Bovinos/inmunología , Calostro/inmunología , Inmunoglobulina M/análisis , Ovinos/inmunología , Animales , Fenómenos Químicos , Química Física , Epítopos , Femenino , Cadenas Pesadas de Inmunoglobulina/análisis , Cadenas Ligeras de Inmunoglobulina/análisis , Inmunoglobulina M/inmunología , Peso MolecularRESUMEN
A low molecular weight antibody which coated Treponema pallidum without producing any apparent reaction was isolated from human syphilitic serum. Its electrophoretic mobility was that of IgG immunoglobulins, and molecular weight was between 115,000 and 120,000.