Diseased Human Kidneys Research Articles

Expression of platelet-derived growth factor receptors in normal and diseased human kidney. An immunohistochemistry and in situ hybridization study.

We studied the expression of PDGF-alpha and -beta receptors in 10 normal and 40 pathologic human kidneys (five minimal change disease, five membranous nephropathy, 25 IgA nephropathy, five lupus nephritis), by both immunohistochemistry and in situ hybridization techniques. In normal-appearing kidneys, both PDGF-alpha and -beta receptors were expressed at the glomerular and interstitial level, the latter receptor more intensely than the former. The distribution and degree of expression of both receptors in nonproliferative glomerulonephritides were comparable with those found in normal-appearing kidneys. PDGF-beta receptor gene and protein expression were upregulated in proliferative nephritides both at the glomerular and the interstitial level and strictly correlated with the grade of histologic lesions. Finally, PDGF beta receptor expression was observed at a low level in normal-appearing renal vessels, and strikingly increased in injured arteries. Diseased kidneys displayed only a slight increase of PDGF-alpha receptor expression, chiefly at the interstitial level. Noteworthy, a few cases of lupus nephritis showed a moderate increase of PDGF-alpha receptor also at the glomerular level. These data establish PDGF-beta receptor activation as a candidate for driving glomerular and interstitial proliferation and, probably, expansion of extracellular matrix in proliferative glomerulonephritis, while the role of PDGF-alpha receptor activation at the renal level remains to be elucidated.

Characterization of a monoclonal antibody recognizing selective epithelial elements of Wilms' tumors and fetal kidneys.

A new antigen was detected using a monoclonal antibody generated against malignant blastema from a Wilms' tumor. This antigen showed variable expression in malignant blastemal cells but was never detected in normal blastema of fetal kidneys irrespective of gestational stage. In a series of 16 Wilms' tumors, the most intense and consistent staining was seen in tubule-associated epithelial cells. Such tubular staining is not surprising as the putative induction of malignant blastema to differentiate into malignant tubules is thought to parallel normal tubulogenesis. This antigen was also associated with epithelial cells located in a variety of fetal kidney structures. Again, the staining was most consistent in tubular epithelia. This monoclonal antibody reactive with a blastemal-epithelial-tubular (BET) antigen should be of value in studying the induction of epithelial differentiation in the normal and diseased human kidney.

Experimental studies for clinical standardization of transabdominal laparoscopic nephrectomy.

In keeping with the maxim that minimal invasiveness is the present working trend in most areas of surgery, we investigated the possibility of laparoscopically removing a diseased human kidney. Before any actual clinical attempt could be made in this direction, an experimental study was set up in order to determine the most beneficial methods and techniques which would guide us through our first clinical cases. This step-by-step training program consists of: (1) initial familiarization with different laparoscopic instruments, bimanual dissection, ligation and coagulation techniques, camera handling and trocar placement as well as organ retrieval systems using the lap simulator; (2) laparoscopic removal of the kidney and ureter of the pig either as short- or long-term study, and (3) laparoscopic simulation with the lap tent during open surgery.

Immunohistochemical localization of C3d fragment of complement and S-protein (vitronectin) in normal and diseased human kidneys: association with the C5b-9 complex and vitronectin receptor.

The localization of C3d, a fragment produced by C3 activation and S-protein (vitronectin), a regulatory factor of C5b-9, was studied immunohistochemically in normal human kidney and renal biopsies from patients with several types of glomerulonephritis. Immunofluorescent staining of the normal kidneys showed that C3d was present along the glomerular basement membrane (GBM), tubular basement membrane (TBM) and arterioles, and that S-protein was present in the GBM, mesangium, TBM, and arterioles. Immunoelectron microscopy of isolated basement membranes showed that C3d was localized exclusively on the epithelial side of the GBM, and that S-protein was present along both the epithelial and endothelial sides. In nephritic tissues, glomerular staining of C3d, C5b-9, and S-protein was increased when compared with that in normal tissues. S-protein, frequently co-localized with C3d and C5b-9 neoantigen, was intensely positive in the immune deposits of glomerular capillaries and the mesangial area, overlapping the background staining of GBM and mesangial matrix. S-protein and its receptor were occasionally co-localized in the glomeruli. These findings indicate that C3d and S-protein are normally present in the glomeruli. Co-staining of C3d, C5b-9 neoantigen, and S-protein within the immune deposits of nephritic kidneys suggests in situ binding of S-protein to locally-formed C5b-9 complex, or merely co-distribution of S-protein with the complex, rather than trapping of large molecular SC5b-9 complex from the circulation.

Expression of fibronectin receptor and vitronectin receptor in diseased human kidneys

Expression of fibronectin and vitronectin receptors was studied in normal human kidney tissues and renal tissue biopsies from patients with several types of glomerulonephritis. Immunofluorescent staining of the normal kidneys showed that fibronectin receptor was present along the glomerular capillary walls, and that vitronectin receptor was present in the vessel walls, but was almost absent from the glomerulus. In kidney tissues biopsied from patients with various renal diseases, expression of fibronectin and vitronectin receptors was correlated with increase in the mesangial expansion. Expression of these receptors was decreased in the sclerosed area and hyalinized glomeruli compared with normal tissues. Fibronectin and vitronectin receptors were occasionally colocalized in the glomeruli with fibronectin and vitronectin, respectively. In situ hybridization showed that fibronectin receptor mRNA and vitronectin receptor mRNA were expressed in the diseased glomeruli. These findings indicate that expression of fibronectin and vitronectin receptors was altered in human glomerulonephritis, and that integrin expression may be important in cell-matrix interaction in the diseased glomeruli.

PAH extraction and estimation of plasma flow in diseased human kidneys

We have analyzed the efficiency with which p-amino-hippuric acid (PAH) is extracted (EPAH) by patients with healthy kidneys (n = 13) or kidneys damaged by chronic cyclosporin nephropathy (n = 21) or primary glomerulopathy (n = 12); respective values (mean +/- SE) for EPAH were 0.87 +/- 0.03, 0.77 +/- 0.03, and 0.69 +/- 0.04. Judged by a 131I-hippuran-to-PAH clearance ratio of 0.75 +/- 0.05, extraction ratio of hippuran was less efficient than EPAH in three glomerulopathic patients. A direct relationship was defined between EPAH and glomerular filtration rate (GFR) (r = 0.54) or calculated efferent oncotic pressure (IIE; r = 0.41, P less than 0.01). Curve fitting by means of quadratic spline functions revealed GFR and IIE to be additive in predicting EPAH (R2 = 0.45). Linear model prediction methods and a sample reuse technique failed to predict EPAH reliably from GFR and preglomerular oncotic pressure (IIA); however, 95% prediction intervals exceed 0.30 EPAH units in width. We conclude that oncotic pressure (presumably reflecting albumin concentration) along with GFR is predictive of EPAH depression in humans with chronic renal disease. However, even sophisticated curve-fitting techniques are too imprecise for accurate prediction of EPAH in a given individual. We submit that renal venous sampling to determine EPAH continues to be necessary for the accurate determination of the rate of plasma flow in the injured human kidney.

Localization of decay accelerating factor in normal and diseased kidneys

Decay accelerating factor (DAF) is a cell membrane associated glycoprotein that inhibits C3 activation. In the present study we evaluated the presence of DAF in normal (N = 15) and diseased human kidneys (N = 76). Sections of frozen tissue were stained for DAF by immunoperoxidase, utilizing three mouse monoclonal anti-DAF anti-bodies. In normal kidneys, DAF was localized in the glomerular vascular pole, apparently in the juxtaglomerular apparatus (JGA). All other structures were negative for DAF. By contrast, in diseased kidneys, two types of abnormalities were detected. First, JGA-DAF was significantly decreased and this abnormality correlated with the pathologic diagnosis and with the presence of C3, IgM and/or fibrinogen in the glomeruli. Second, DAF was present in the glomerular mesangium (67%), renal interstitium (68%) and/or blood vessels (38%). The presence of DAF in the mesangium and interstitium of the kidney correlated with each other and correlated with C1q and C3 deposition in the glomerulus. Finally, vascular DAF was significantly more common in patients with electron dense deposits in the glomeruli. In summary, DAF is present in the normal kidney and is located exclusively in the glomerular vascular pole. In diseased kidneys, DAF tends to be lost from the JGA but is often present in glomerular mesangium, interstitium and blood vessels. This pattern is specially prominent in patients demonstrating complement deposition in the glomerulus. We speculate that kidney DAF may play a role in protecting the kidney against the products of complement activation.

Lectin Binding Patterns of Normal and Diseased Human Kidneys as Revealed by Histochemical Techniques

Lectin Binding Patterns of Normal and Diseased Human Kidneys as Revealed by Histochemical Techniques

Immunohistochemical study of Ia antigen in the normal and diseased human kidney

AbstractImmunohistochemical study of Ia antigen in the normal and diseased human kidney. The presence and distribution of Ia antigen in normal human kidneys and biopsy specimens from patients with renal disease were investigated by immunohistochemical techniques using two monoclonal antibodies to the nonpolymorphic determinants of human HLADR molecules. Ia antigen was found on the endothelium of glomerular and peritubular capillaries and of veins and vasa recta. Loss of endothelial staining was found in necrotic and sclerotic glomerular and tubulointerstitial lesions. Staining or decreased staining was also not found in the severe proliferative nephritis of systemic lupus erythematosus although endothelial cells could still be identified upon light microscopic examination of the same biopsy specimen. Resting and proliferating cells in mesangial areas did not stain with anti-Ia antibody and extracapillary proliferating cells did not express Ia antigen except for occasional cells in anti-GBM crescentic glomerulonephritis, suggesting that Ia-bearing cells are not involved in mesangial and most extracapillary proliferations in human glomerulonephritis. All clustered mononuclear cells infiltrating the renal interstitium stained with anti-Ia antibody regardless of the type of nephritis where infiltrates occurred.Etude immunohistochimique de l'antigène Ia dans les reins humains normaux et pathologiques. La présence et la distribution de l'antigène Ia dans les reins humains normaux et dans les spécimens biopsiques de patients porteurs de néphropathies diverses ont été étudiées par des techniques immunohistochimiques à l'aide de deux anticorps monoclonaux dirigés contre les déterminants non polymorphiques des molécules HLA-DR humaines. L'antigène Ia a été décelé sur l'endothelium des capillaires glomérulaires et péritubulaires des veines et des vasa recta. Une perte de marquage endothélial a été constatée dans les lésions nécrotiques et scléreuses glomérulaires et tubulointerstitielles. Une absence de marquage ou un marquage atténué ont aussi été observés dans les glomérulonéphrites prolifératives graves du lupus érythémateux disséminé bien que les cellules endothélials pouvaient être encore identifiées en microscopie optique sur le même spécimen biopsique. Les cellules présentes dans l'aire mésangiale, qu'elles soient en nombre normal ou accru, n'étaient pas marquées par l'anticorps anti-Ia. Les cellules extracapillaires proliférées n'exprimaient pas l'antigène Ia, excepté quelques cellules dans le croissant des glomérulonéphrites extracapillaires. Ces faits suggèrent que les cellules porteuses de l'antigène la ne sont pas concernées dans les proliférations mésangiales ou extracapillaires au cours des glomérulonéphrites humaines. Toutes les cellules mononuclées en amas qui infiltraient l'interstitium rénal étaient marquées avec l'anticorps anti-la quelle que soit la néphropathie en cause.

Increased concentration of albumin in kidney basement membranes in diabetes mellitus.

The concentration of albumin was measured by a double-antibody radioimmunoassay in the glomerular basement membrane (GBM) and tubular basement membrane (TBM) of patients with diabetic nephropathy, other kidney diseases, and normal kidneys. The TBM from diabetics contained significantly more albumin (m +/ SD; 2.75 +/- 0.34 microgram/mg) than the TBM from other diseases (1.00 +/- 0.24 microgram/mg) or normals (1.21 +/- 0.26 microgram/mg) (P less than 0.0001). Similarly, diabetic GBM contained more albumin (2.25 +/- 0.59 microgram/mg) than other diseases (1.22 +/- 0.55) or normals (1.31 +/- 0.36 microgram/mg) (P less than 0.01-0.001). No differences were observed between the normal and other disease groups. Although there were no differences in hydroxyproline content, there was a highly significant correlation between the concentration of hydroxyproline and albumin in the diabetic TBM (r = 0.82), diabetic GBM (r = 0.70), and normal TBM (r = 0.79). Elution studies with different buffers on frozen sections of diabetic kidneys suggest that the albumin is relatively firmly bound. Although albumin is present in extracellular membranes of normal and diseased human kidneys, the concentration is higher in diabetes mellitus.

Distribution of fibronectin in normal and diseased human kidneys.

The distribution of fibronectin in 42 renal biopsies was investigated using an indirect immunoperoxidase technique on paraffin sections of formalin-fixed material. Biopsies were obtained from normal human kidneys and from patients with a variety of glomerular disorders. In normal glomeruli, fibronectin was present in Bowman's capsule, in the mesangium, and outlining peripheral capillary loops. A significant increase in fibronectin was observed in most types of glomerulopathy with a distribution closely related to the histopathological diagnosis. Fibronectin was diminished or absent in segmental scars, old diabetic nodules, and hyalinised glomeruli.

Cold-insoluble globulin (fibronectin, LETS protein) in normal and diseased human glomeruli: Papain-sensitive attachment to normal glomeruli and deposition in crescents

Cold-insoluble globulin (Clg, fibronectin, LETS protein) is a major plasma protein of unkown function. Its ability to interact with collagen, fibrin, and cell surfaces has prompted this study of Cig in normal and diseased human kidneys. CIg was detected by indirect immunoflorescence in the normal glomerular mesangium and basement membrane, in the tubular basement membrane, in vessel walls, and in the interstitium. Isolated glomerular matrix was able to absorb anti-CIg antibodies, even when treated with 8 M urea, 3 M KSCN, nonionic detergents, EDTA, and dithiothreitol. In contrast, papain was able to remove CIg from glomerular matrix, but did not remove CIg linked to Sepharose. CIg was also released by complete solubilization of glomerular matrix with collagenase and detected by gel diffusion. These results indicate that CIg is strongly attached to the normal human glomerulus, probably by covalent nonsulfhydryl bonds. In the 27 cases of renal diseases examined, mesangial CIg was increased in all 8 of those with increased mesangial matrix (diabetes mellitus, membranoproliferative glomerulonephritis type I, IgA nephropathy and Henoch-Schönlein purpura). CIg was not a component of amyloid deposits (2 cases). Eleven cases of glomerulonephritis with crescents were studied. Each had prominent deposition of CIg between the proliferating cells in the crescents in association with fibrin/fibrinogen. Because CIg is known to bind to fibrin and to cells as well as to collagen, we suggest that CIg may play a pathogenetic role in the formation of cellular crescents and their subsequent evolution into fibrous scars.

The concentration of thiamphenicol in severely diseased human kidneys.

The concentration of thiamphenicol in serum and renal tissue was determined in 17 patients with severly diseased kidneys after an intravenous injection of 1000 mg of the drug. Two hours after the administration the renal tissue concentrations ranged in patients with hydronephrotic kidneys from 38.0-63.5 microgram/g, in patients with cirrhotic kidneys from 17.9-42.7 microgram/g, in patients with pyonephrosis from 9.8-17.6 microgram/g and in patients with renal carcinoma from 37.7-64.2 microgram/g. The patient with the renal sarcoma had a level of 138.7 microgram/g. At the same time the serum concentration ranged from 4.6-15.2 microgram/ml. The highest renal tissue/serum concentration ratios of thiamphenicol were observed in patients with hydronephrotic kidneys and renal tumours, the lowest in cases of pyonephrosis. The influence of severe renal disease on the renal tissue/serum concentration ratios of thiamphenicol is discussed. The high renal tissue levels of thiamphenicol in patients with severely diseased kidneys fulfill an important condition for the antibacterial chemotherapy of kidney infections.

Measurement of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase activity in human tissues.

The activity of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (aspartylglucosylaminase, EC 3.5.1.26) was measured in normal and diseased human liver, brain and kidney. Organs from patients with aspartylglucosaminuria show very little activity. Crude homogenates of human organs show a reaction catalysed by a complex enzyme system. With homogenate, the formation of product was linear with time up to about 6 h. Reaction times longer than 6-7h resulted in a decrease in the total concentration of product. This phenomenon was not found with the partially purified enzyme fraction. Linearity of the enzyme activity with different protein concentrations was found, independent of the incubation time. Longer incubation of the crude homogenate resulted in the utilization of the product, N-acetylglucosamine. This phenomenon was not observed with the partially purified enzyme fraction. This amidase from human organs differs from that obtained from other sources and apparently represents a rather complex enzyme system.

Pharmacokinetic Characteristics of Doxycycline Acwmulation in Normal and Severely Diseased Kidneys

Intrarenal concentrations of doxycycline were measured in 48 normal canine kidneys during wide variations in urine pH and the systemic state of hydration of the experimental animals. The results were compared with the renal parenchymal levels achieved in 10 severely diseased human kidneys. Although the state of hydration, the urine pH, and renal disease all significantly influenced the urinary concentrations and rate of renal clearance of doxycycline, there was no detectable difference between concentrations in normal renal tissue and those in severely diseased kidneys. This result is in contradistinction of the findings for previously evaluated antibiotics. The levels of doxycycline measured in renal tissue averaged twice the concentration in serum without notable difference among levels in renal cortex, medulla, and papilla. Prospective clinical trials will be necessary for identification of any solid correlation between the remarkable characteristics of accumulation of doxycycline in diseased renal parenchymal tissue and the clinical, therapeutic importance of such an observation.

Synthesis of Immunoglobulins and 15 Other Proteins by Diseased Human Kidneys*

Summary. In an effort to determine the synthetic capacity of human kidneys for serum proteins, pieces of both kidneys removed at operation prior to transplantation were placed in organ culture. Incubation with 14C labelled amino acids and subsequent radioimmunoelectrophoresis with antisera to all five classes of immunoglobulins showed synthesis of IgG by all 5 pairs of kidneys of IgA and IgE by 4, and of IgM by 2. None synthesized IgD. Radioimmunoelectrophoresis with specific antisera to 15 non‐immunoglobulin proteins revealed synthesis of lactoferrin and uromucoid while several other serum proteins, such as α1 antitrypsin and α2 macroglobulin, known for their binding properties, showed apparent synthesis. No evidence for secretory component synthesis was seen despite positive immunofluorescent localization in the tubular cells. Further, Zn α2 and b̊2GP also showed positive immunofluorescence but negative synthesis. It seems possible that these latter proteins may not be synthesized by the tubular cells but rather may be actively excreted or reabsorbed by them.

Synthesis of immunoglobulins and 15 other proteins by diseased human kidneys.

In an effort to determine the synthetic capacity of human kidneys for serum proteins, pieces of both kidneys removed at operation prior to transplantation were placed in organ culture. Incubation with 14C labelled amino acids and subsequent radioimmunoelectrophoresis with antisera to all five classes of immunoglobulins showed synthesis of IgG by all 5 pairs of kidneys of IgA and IgE by 4, and of IgM by 2. None synthesized IgD. Radioimmunoelectrophoresis with specific antisera to 15 non-immunoglobulin proteins revealed synthesis of lactoferrin and uromucoid while several other serum proteins, such as alpha1 antitrypsin and alpha2 macroglobulin, known for their binding properties, showed apparent synthesis. No evidence for secretory component synthesis was seen despite positive immunofluorescent localization in the tubular cells. Further, Zn alpha2 and beta2GP I also showed positive immunofluorescence but negative synthesis. It seems possible that these latter proteins may not be synthesized by the tubular cells but rather may be actively excreted or reabsorbed by them.

Application of HVEM to Thick Sections of Diseased Human Kidney

The high voltage transmission electron microscope (HVEM) offers the biologist a means of examining ultrastructure in thick (0.3 to 1.0μ) sections of conventionally enbedded biological material with good Z-axial resolution. The spatial distribution of structures throughout the depth of a specimen may be easily visualized by HVEM and the need for reconstruction from ultrathin serial sections avoided. Effective stereo pair micrographs allowing the 3-D relationships of organellae to be observed are also readily made.Our preliminary applications of HVEM in the examination of diseased human kidney were made in an effort to gather information on poorly understood renal pathologic changes. These changes include loss or alteration in size of glomerular epithelial and endothelial cells and their processes, changes in thickness or composition of glomerular basement membrane, the deposition of subepithelial or subendothelial electron-dense material, and the occurrence of round or microtubular virus-like particles and crystalline deposits.

Intrarenal distribution of ampicillin in the normal and diseased human kidney.

In the normal human kidney, the intrarenal tissue concentrations of ampicillin in the hydrated and hydropenic state demonstrate a pattern similar to that previously described for the kidney of the dog. The outer medullary and papillary concentrations, of presumed importance in the treatment of pyelonephritis, are uninfluenced by the state of hydration. In the severely diseased human kidney, a marked reduction in tissue penetration of the antibiotic is noted, and the ability of the kidney to concentrate ampicillin in the urine is greatly reduced. Levels in serum are almost twice as high as the levels in renal tissue. Studies of intrarenal distribution of ampicillin in the normal and diseased state should facilitate interpretation of tube-dilution studies in the management of complicated renal infections.

Redistribution of intrarenal blood flow in the patients with chronic renal disease.

In order to study the distribution of blood flow in diseased human kidneys, Xe133 washout curves were obtained from 34 patients with chronic renal disease or hypertension.The curves recorded for over 45min. were resolved into 4 components, while the curves recorded for 30min. showed 3 components. The values of components I, II and III in the 30min. record coincided with those of the components I, the average of II and III, and IV in the 45min. record, respectively.The percentage of the total renal blood flow (flow ratio) distributed to the component I of the 30min. record was correlated with that due to cortical blood flow as determined by the dye-dilution method. It is suggested that the component I of the Xe133 washout curve from the human kidney represents the cortical blood flow.The results obtained from the Xe133 washout curves recorded for 30min. were compared with the renal function of the patients. The flow for unit mass of tissue (flow rate) and the flow ratio of the fastest component (component I) were decreased in the azotemics. The flow rate of component I was also decreased in proportion to the reduction of the RBF calculated from CPAH (RBFPAH), GFR from CThio and maximal urinary osmolality, while the flow rate of component II did not change significantly. As the renal function became impaired to the level of RBFPAH less than 500ml./min., GFR 50ml./min. and maximal urinary osmolality 700mOsm, the flow ratio of component I decreased and that of component II increased progressively.