[show abstract][hide abstract] ABSTRACT: Glomerular visceral epithelial cells (GVEC) from normal human glomeruli were grown in tissue culture. Cell surface markers were studied by immunofluorescence microscopy using antibodies against lymphohaemopoietic differentiation antigens which are known to be present early (BA-1, OKB2, BA-2) and late (J5, anti CR1) in renal ontogenesis. Like foetal human glomerular epithelium, the cultured cells reacted with BA-1 and OKB2 (identifying an antigen expressed on B cells and polymorphonuclear leucocytes), and BA-2 (leukaemia-associated antigen), but were consistently negative for CR1 (C3b receptor); J5 which identifies the common acute lymphoblastic leukaemia antigen (CALLA) stained variably. Reactivity with antimyosin or anti factor VIII were absent. The cells produced an extracellular matrix containing laminin, type IV collagen, and fibronectin. This study supports the notion that GVEC undergo dedifferentiation as shown by the acquisition of lymphohaemopoietic differentiation antigens present early in renal ontogeny. In addition, the production of extracellular matrix constituents in vitro may be useful for the investigation of human glomerular basement membranes.
British journal of experimental pathology 03/1989; 70(1):73-82.
[show abstract][hide abstract] ABSTRACT: The presentation, clinical course, and long-term follow-up (3-22 years) of 39 children with dermatomyositis followed from 1962-1982 is presented. The medical course of these patients was complicated by respiratory diseases (20%), gastrointestinal diseases (24%), and calcinosis (30%). Patients presenting prior to 1972 received a wide variety of treatments. Since 1972, treatment has consisted of long-term prednisone, supplemented in some patients with azathioprine. Ten patients died (eight of whom were first seen before 1972) of complications after intestinal perforation (5) or aspiration pneumonia (5). Of 29 survivors, three have persistent calcinosis and/or contractures. Improved outcome since 1972 probably relates to better clinical assessment, management of complications, and regulation of drug therapy.
[show abstract][hide abstract] ABSTRACT: Utilizing a monoclonal antibody (Poly C9-MA) to a neoantigen of the C9 portion of the membrane attack complex of complement (MAC), immunoelectron (IEM) and immunofluorescent (IF) microscopy were performed on kidney tissue from normal humans and patients with insulin-dependent diabetes mellitus (IDDM) and type II membrano-proliferative glomerulonephritis (MPGN II). Comparative studies were conducted using polyclonal antibodies to human C3, C5, IgG, IgA, and IgM. In normal human tissue, there was a close correlation between increasing chronologic age and the quantity of MAC deposited in the mesangial stalk, along the interstitial aspect of and within tubular basement membranes (TBMs) and in arteriolar walls. IF of kidney tissues from 12 patients with IDDM with varying degrees of mesangial expansion and glomerulosclerosis demonstrated a direct relationship between the degree of tissue damage and the amount of MAC deposited in the mesangium. IEM of three normal and four diabetic specimens revealed reaction product of Poly C9-MA on linear and circular membranous structures within the mesangium, TBMs, and vessel walls, and within the glomerular basement membranes (GBMs) in diabetic subjects. Evidence is presented that these structures, which have been previously described by routine electron microscopy, represent cellular debris in these loci on which Poly C9-MA has been deposited. In MPGN II, Poly C9-MA and C3 were distributed within subepithelial deposits, along either side of the dense deposits (DDs) within the GBMs and TBMs, and around circular masses of DDs within the mesangium.(ABSTRACT TRUNCATED AT 250 WORDS)
American Journal of Kidney Diseases 03/1987; 9(2):121-8. · 5.29 Impact Factor
[show abstract][hide abstract] ABSTRACT: Serum amyloid P component (SAP) is a normal plasma glycoprotein immunochemically indistinguishable from amyloid P, a glycoprotein found in all tissue amyloid deposits. SAP has also been shown to be a constituent of normal glomerular basement membrane (GBM). In this study the authors discovered a unique association between SAP and Goodpasture (GP) antigen. In those patients whose GBM lack GP antigen (those with Alport-type hereditary nephritis) SAP is also uniformly absent. Although the relationship between these two components is unknown, this association may provide clues to the abnormality of GBM in Alport-type hereditary nephritis.
American Journal Of Pathology 01/1987; 125(3):460-4. · 4.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: We developed a radioimmunoassay to measure the attack complex of complement (SC5b-9) in serum from patients with systemic lupus erythematosus. The radioimmunoassay used a monoclonal antibody to an antigen of C9, which is absent from native C9 but present on SC5b-9. SC5b-9 was detectable in 13 of 63 normal subjects, with a mean value of 0.5 unit (range, 0 to 4.8), and elevated in 13 of 14 patients with active lupus. Analysis of 108 samples from the patients with lupus revealed a mean value of 10.1 units (range, 0 to 35) when the disease was active and 1.0 unit (range, 0 to 433) when it was clinically stable. SC5b-9 was a more sensitive measure of disease activity than C3, C4, or CH50; its specificity was equivalent to that of C3 and C4. SC5b-9 was elevated in serum from 6 of 49 patients with other forms of glomerulonephritis. Our study documents the presence of SC5b-9 in abnormal serum and correlates its elevation with clinical disease activity in patients with active systemic lupus erythematosus.
New England Journal of Medicine 07/1985; 312(25):1594-9. · 51.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Renal tissue from 9 patients with diabetes mellitus (4 with mild and 5 with end-stage disease) and 3 with antiglomerular basement membrane (GBM) nephritis, as well as 5 normal human kidneys, were examined by immunofluorescence microscopy for the presence of plasma proteins of varying isoelectric point (pI). In normal and diabetic kidneys, IgG deposition in basement membranes was restricted to IgG4 (pI 5.5-6.0), the subclass present in lowest concentration in human plasma. IgG1, IgG2, and IgG3 (pI 7.0-9.5) were not detected. In contrast, in anti-GBM nephritis, all four subclasses were present in a linear pattern in GBM. Other plasma proteins of low isoelectric point were detected in basement membranes: albumin (pI 4.9), alpha-1-acid glycoprotein (pI 2.7), amyloid P (pI 3.9-4.8), and alpha-1-antitrypsin (pI 4.5). These studies are consistent with the hypothesis that circulating anionic plasma proteins are electrostatically bound in vivo to positively charged moieties in normal and especially diabetic basement membranes.
American Journal Of Pathology 07/1984; 115(3):443-6. · 4.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: Dense intramembranous deposit disease (DIDD) almost universally recurs in renal allografts. However, the 29 previously reported cases suggest that recurrent DIDD rarely results in graft failure. We studied the clinical course and renal histology of the 6 patients with DIDD who have received renal allografts here since 1967. All patients had characteristic findings of DIDD in their native kidneys by light, immunofluorescence and electron microscopy. Seven grafts in 4 patients failed and histological evaluation showed that 5 of these allografts in 4 patients were lost due to recurrent disease. They all demonstrated marked mesangial proliferation with crescents but minimal acute interstitial or vascular changes of rejection. Patients with graft failure due to recurrent disease were male and developed recurrent nephrotic syndrome. The 2 patients with rapidly progressive glomerulonephritis (RPGN) in their native kidneys lost the transplanted kidney due to recurrent disease within 1 year. This study suggests that recurrence of DIDD in renal allografts is a more serious problem than previously appreciated, especially in patients with RPGN.
[show abstract][hide abstract] ABSTRACT: The possibility that experimental diabetes could prepare for the generalized Shwartzman reaction was investigated in female Sprague-Dawley streptozotocin-induced diabetic rats. After 48 hours, 1 week, and 9 weeks of diabetes, the rats were injected with 2 mg/kg of endotoxin, and the animals were sacrificed 2, 4, 8, and 24 hours after endotoxin. Ninety percent of the diabetic animals given endotoxin developed massive glomerular capillary fibrin deposition accompanied by marked decrease in platelet count. The age- and sex-matched nondiabetic control rats had no such changes. This marked susceptibility to endotoxin, previously only reported in pregnant rats, was present as early as 1 week of diabetes. The degree of glycemic control greatly influenced the susceptibility of diabetic rats to the generalized Shwartzman reaction. Only 28% of the diabetic animals given insulin once daily (4.6 +/- 0.3 units, mean +/- SEM) and maintaining a blood glucose level of 269 +/- 19 mg/dl developed glomerular thrombi. In contrast, the diabetic animals that did not receive insulin and had a blood glucose level of 617 +/- 21 mg/dl all developed fibrin thrombi. We conclude that the diabetic state in rats induces a unique susceptibility to the generalized Shwartzman reaction following a single injection of endotoxin, which varies with the severity of the diabetic state. Although the pathogenesis is unclear, this phenomenon may reflect abnormalities in the glomerular capillary wall and/or the coagulation system that may be important in the development of microvascular complications. Furthermore, this phenomenon may, in the animal model, mirror the increased risk of the diabetic patient to intravascular coagulation with bacterial sepsis.
[show abstract][hide abstract] ABSTRACT: The kinetics of radiolabeled heat-aggregated human IgG (AHIgG125I) were studied in rats with passive Heymann's nephritis (PHN) induced 72 hr previously with decomplemented rabbit antiserum to rat FX1A. Control rats were injected with decomplemented normal rabbit serum (NRS). Following administration of AHIgG125I (40 mg per 100 g of body wt) control and FX1A animals were sacrificed in groups of five each at 2, 4, 8, 16, and 24 hr and kidney, liver, spleen, lung, plasma, and blood cells obtained. 131I-Labeled human serum albumin (HSA131I) was administered prior to sacrifice as a plasma marker. In FX1A rats the following observations were made in comparison with control rats: (1) A decrease in the concentration of AHIgG125I in glomeruli was observed at 2, 4, and 8 hr after administration; (2) a significant increase in clearance reflected by a decrease in the concentration of plasma trichloroacetic acid (TCA)-precipitable radioactivity, and AHIgG125I (greater than 7 S) was present; (3) a significant increase in non-TCA-precipitable radioactivity in plasma and blood cells at most time periods; and (4) decreased concentrations of AHIgG125I in liver and spleen but not lung. The specificity of these observations was supported in separate experiments by the lack of any difference in the plasma levels of TCA-precipitable radioactivity after administration of radiolabeled albumin to FX1A and control rats. Studies in FX1A and control rats revealed no differences in body weight, kidney weight, hematocrit, blood volume, urine output, glomerular filtration rate, renal blood flow, or renal vascular resistance. A slight increase in urinary rat albumin excretion was observed in FX1A rats. The lower values of AHIgG125I observed in plasma, liver, and spleen associated with increased levels of non-TCA-precipitable radioactivity in plasma and blood cells suggest enhanced catabolism of AHIgG125I in FX1A rats, leading to decreased localization within the mesangium.
Clinical Immunology and Immunopathology 04/1984; 30(3):393-404.
[show abstract][hide abstract] ABSTRACT: A monoclonal antibody to a neoantigen of the C9 portion of the membrane attack complex (MAC) of human complement has been developed and characterized. The distribution of this neoantigen was assessed by indirect immunofluorescence microscopy in nephritic and nonnephritic renal diseases. The antibody (Poly C9-MA) reacted on enzyme-linked immunosorbent assay (ELISA) with a determinant in complement-activated serum that was undetectable in normal human serum (NHS). Zymosan particles incubated in NHS had positive immunofluorescent staining with Poly C9-MA; however, binding of Poly C9-MA was not observed with zymosan particles incubated in sera deficient in individual complement components C3, C5, C6, C7, C8, or C9. Reconstitution of C9-deficient sera with purified C9 restored the fluorescence with Poly C9-MA. Poly C9-MA reacted positively by ELISA in a dose-dependent manner with purified MC5b-9 solubilized from membranes of antibody-coated sheep erythrocytes treated with NHS but not with intermediate complement complexes. Poly C9-MA also reacted in a dose-dependent manner on ELISA and in a radioimmunoassay with polymerized C9 (37 degrees C, 64 h) (poly C9) but not with monomeric C9. Increasing amounts of either unlabeled poly C9 or purified MC5b-9 inhibited the 125I-poly C9 RIA in an identical manner. These studies demonstrate that Poly C9-MA recognizes a neoantigen of C9 common to both the MAC and to poly C9. By immunofluorescence, Poly C9-MA reacted minimally with normal kidney tissue in juxtaglomerular loci, the mesangial stalk, and vessel walls. Poly C9-MA stained kidney tissue from patients with glomerulonephritis in a pattern similar to that seen with polyclonal anti-human C3. In tissue from patients with nonnephritic renal disease--diabetes, hypertension, and obstructive uropathy--Poly C9-MA was strongly reactive in the mesangial stalk and juxtaglomerular regions, tubular basement membranes, and vascular walls. Poly C9-MA binding was especially prominent in areas of advanced tissue injury. Poly C9-MA frequently stained loci where C3 was either minimally present or absent. These studies provide strong evidence for complement activation not only in nephritic but also in nonnephritic renal diseases.
Journal of Clinical Investigation 09/1983; 72(2):560-73. · 12.81 Impact Factor
[show abstract][hide abstract] ABSTRACT: Polymeric IgA and polymeric IgA-containing immune complexes are transported from blood to bile through hepatocyte-bound secretory component. In order to investigate interruption of this transport and its effect on the glomerulus, Sprague-Dawley rats underwent bile duct ligation. Renal tissue obtained at the time of sacrifice was stained by immunofluorescent techniques with antibodies to IgG, IgM, and IgA, and secretory component (SC) and C3. A progressive selective increase in the staining intensity of the glomerular mesangium was observed for IgA, C3, and SC in bile duct ligated rats. These changes were paralleled by a rise in serum IgA and C3. These findings are consistent with the view that glomerular deposition of IgA, C3, and SC in bile duct ligated rats may result from impairment of normal handling of polymeric IgA and polymeric IgA-containing immune complexes.
Clinical Immunology and Immunopathology 07/1983; 27(3):369-77.
[show abstract][hide abstract] ABSTRACT: The effects of prostacyclin infusion on endotoxin-induced generalized Shwartzman reaction were studied in pregnant rats. Four hours after administration of Escherichia coli endotoxin (0.4 mg) to pregnant rats on the 20th day of gestation, fibrin and platelet thrombi were observed in 72% of the animals. These findings were associated with a decrease in renal blood flow and glomerular filtration rate from 7.0 to 0.7 and 0.8 +/- 0.1 to 2.6 +/- 0.4 and 0.2 +/- 0.8 ml/minute, respectively (p less than 0.0001 and less than 0.05) and a decrease in platelet count (p less than 0.05). The infusion of prostacyclin, at a mean dose of 57 ng/kg/minute prior to and after administration of endotoxin strikingly inhibited the development of fibrin and platelet thrombi in glomerular capillaries (p less than 0.001). Despite hemodynamic changes which were similar to those in control animals, renal blood flow and glomerular filtration rate decreased from 7.2 +/- 0.2 and 0.9 to 2.9 +/- 0.03 and 0.3 +/- 0.1 ml/minute, respectively (p less than 0.001 and less than 0.025). In addition, the mean blood pressure decreased from 118 +/- 3.8 to 86 +/- 3.7 mm Hg (p less than 0.025), whereas the platelet count remained normal. We conclude that prostacyclin infusion prior to endotoxin administration significantly inhibits endotoxin-induced generalized Shwartzman reaction in pregnant rats.
[show abstract][hide abstract] ABSTRACT: The presence of Goodpasture's (GP) antigen in the glomerular basement membrane (GBM) of the kidney was evaluated by indirect immunofluorescence in nine patients with familial nephritis from five kindreds. The GP antigen was not detected in seven males but was present in an affected sister and mother, an unaffected brother, and 13 normal controls. The specificity of this finding in affected males is supported by the persistence of other GBM antigens identified by monoclonal antibodies. The lack of GP antigen in affected males and its persistence in related females with the disease suggests a possible X-linked dominant mode of inheritance. We propose that the absence of GP antigen leads to severe disease in the male, whereas its presence in related females is associated with mild disease.
American Journal of Kidney Diseases 06/1983; 2(6):626-9. · 5.29 Impact Factor
[show abstract][hide abstract] ABSTRACT: Phagocytic cells provide the host its major defense against invasive Staphylococcus aureus, and the staphylococcal surface, by its influence on phagocyte recognition, is a primary determinant of the function of these cells. The peptidoglycan component of the cell wall plays a key role in both opsonic and chemotactic recognition, mediated by IgG, C3b, and C5a, respectively. While cell wall protein A inhibits opsonic recognition by polymorphonuclear leukocytes, it promotes an opsonin-independent mechanism of phagocytosis by human macrophages which possess cytophilic IgG. By masking cell wall-associated opsonic molecules, capsular polysaccharides inhibit recognition, a phenomenon that is overcome by specific anti-capsular antibodies. It is proposed that impaired phagocyte recognition is a basic element in the pathogenesis of staphylococcal endocarditis, and progress in the prevention and treatment of this infection may depend on understanding the basis for this host defense defect.
Scandinavian journal of infectious diseases. Supplementum 02/1983; 41:67-78.
[show abstract][hide abstract] ABSTRACT: Twenty-six children with hemolytic-uremic syndrome (HUS) had radionuclide studies while oliguric or anuric in order to assess prognosis. Renographic abnormalities were compared with clinical and biochemical parameters and eventual outcome. Presence of an anephric pattern failed to predict development of irreversible renal damage. The authors conclude that radionuclide renography is of no value in children with HUS.
[show abstract][hide abstract] ABSTRACT: The alpha-toxin (hemolysin) of Staphylococcus aureus is known to be an important determinant of pathogenicity although its precise role in the process of infection is not understood. In this study, the interaction of alpha-toxin with the human complement system was evaluated in terms of its effect on the opsonic activity of serum for S. aureus. Phagocytosis by human polymorphonuclear leukocytes was studied by measuring the uptake of preopsonized radiolabeled bacteria. It was found that alpha-toxin-treated serum had reduced opsonic activity and that this change was associated with complement consumption via the classical pathway. Levels of C3 to C9 were reduced in proportion to the amount of toxin added to the reaction mixture; levels of C2 were markedly reduced but those of factors B and D of the alternative pathway were unaltered in the presence of alpha-toxin. Heat-inactivated toxin, which had no hemolytic activity, also interacted with the complement system but with a significantly reduced effect. In addition, alpha-toxin behaved as a chemotaxinogen for polymorphonuclear leukocytes: human serum was activated by the toxin. These studies demonstrate that the interaction of staphylococcus alpha-toxin with human serum affects two important aspects of the host response to the staphylococcus.
Infection and Immunity 01/1983; 38(3):981-5. · 4.07 Impact Factor
[show abstract][hide abstract] ABSTRACT: Previous studies of encapsulated Staphylococcus aureus have shown that the opsonins of normal, nonimmune human serum (complement factor C3 and IgG) bind beneath the capsule, i.e., on the cell wall, and when bound at this site these opsonins are not effective in promoting phagocytosis of the bacteria by polymorphonuclear leukocytes (PMN). In this investigation immune antibody was added to human serum to effect opsonization of encapsulated S. aureus. Opsonization was assessed by quantitating the uptake of 3H-labeled staphylococci by human PMN, and the amount of C3 fixation to bacteria was measured in a quantitative fluorescent immunoassay. Low levels of immune antibody (IgG) effectively opsonized encapsulated S. aureus when added to fresh but not to heated serum; phagocytosis of the staphylococci was mediated via pronase-sensitive membrane receptors (presumably C3b receptors) of PMN. Experiments with C2-, C3-, or C5-deficient human sera revealed that C3 was required for opsonization and that activation of C3 was mediated via the alternative complement pathway. Encapsulated S. aureus bound significantly less C3 than unencapsulated strains in diluted normal serum; addition of immune antibody, however, increased C3 fixation 4.7-fold (p less than 0.005). Immunoelectron microscopy localized C3 throughout the capsule as well as on the staphylococcal cell wall when bacteria had been opsonized in human serum with immune antibody. Without immune antibody, C3 binding was restricted to the cell wall. At approximately 10-fold higher levels of immune antibody, opsonization and phagocytosis of encapsulated S. aureus was independent of complement and pronase-sensitive receptors on PMN. These studies show that, in addition to immune antibody, the alternative pathway of complement plays an important role in the opsonization of encapsulated S. aureus strains and suggest that complement may be crucial to the in vivo clearance of these organisms.
The Journal of Immunology 11/1982; 129(4):1681-7. · 5.52 Impact Factor
[show abstract][hide abstract] ABSTRACT: Seventy patients with systemic lupus erythematosus (SLE) in the first two decades of life have been followed from 1958 to 1981. The patients were treated with corticosteroids alone (17 patients) or corticosteroids and immunosuppressives (53 patients). Twenty-one patients (30%) had evidence of CNS lupus. Eighty-seven percent of patients had evidence of renal involvement, and 34% of these patients had diffuse proliferative lupus nephritis (DPLN). A high frequency of pathologic transition was found to occur. Analysis of 20 pregnancies in 12 women with SLE reveals that patients with active SLE and/or renal disease are at a high risk for complications of pregnancy in contrast to patients in remission. The occurrence of infection, aseptic necrosis and cataracts is shown to be related to SLE itself and to its treatment. By life table analysis overall survival was 90% at 1 yr, 85% at 10 yr. and 77% at 15 yr. Survival of patients who entered the study before age 16 and of patients with CNS lupus could not be distinguished from that of the total group. Patients who entered the study from 1974-1981 had 5-yr survival similar to that of the total group. Survival of 21 patients with DPLN did not deviate from that of the entire group until 14 yr after study entry (7 yr if dialysis and transplantation are considered as deaths). In conclusion, the prognosis of SLE and especially of DPLN in the younger patient appears to be better than indicated by prior reports. Thus, analysis of outcome in this disease may require a long duration of follow-up.
American Journal of Kidney Diseases 08/1982; 2(1 Suppl 1):212-22. · 5.29 Impact Factor
[show abstract][hide abstract] ABSTRACT: In this report, the mechanism of resistance to phagocytosis by group A streptococci and the participation of M protein in this process were studied. A quantitative fluorometric immunoassay was used to determine the amount of the opsonic third component of complement, C3, fixed by group A streptococci in the presence and absence of M protein. In the absence of M protein, phagocytosis was found to be dependent on the amount of C3 fixed. However, when M protein was present, the streptococci still bound sufficient C3 to have promoted phagocytosis, yet none was observed. Further investigation using immunofluorescent microscopy demonstrated that the C3 fixed to M+ chains was confined to dense patches, interspaced by areas devoid of detectable C3. This uneven distribution of C3 was dependent on the presence of a trypsin-sensitive structure; most likely the M protein. The significance of this restricted C3 deposition with respect to resistance to phagocytosis by group A streptococci is discussed.
The Journal of Immunology 05/1982; 128(4):1897-902. · 5.52 Impact Factor