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Biomedical subjects

A J Fish

Publications and source records attributed to A J Fish.

At least 37 records · Page 2Linked to original sources
PubMed1987

Alport familial nephritis. Absence of 28 kilodalton non-collagenous monomers of type IV collagen in glomerular basement membrane.

Alport-type familial nephritis (FN), a genetic disorder, results in progressive renal insufficiency and sensorineural hearing loss. Immunochemical and biochemical analyses of the non-collagenous (NC1) domain of type IV collagen isolated from the glomerular basement membranes (GBM) of three males with this disease demonstrate absence of the normally occurring 28-kilodalton (kD) NC1 monomers, but persistence of the 26- and 24-kD monomeric subunits derived from alpha 1 and 2 (both type IV) collagen chains, respectively.

Basement Membrane↗
PubMed1986

Antibody specificity of human glomerular basement membrane type IV collagen NC1 subunits. Species variation in subunit composition.

NC1 subunits were purified from gel filtration pools of acid-extracted, collagenase-digested human glomerular basement membranes (hGBM). This methodology, which enriches 28-kDa monomers (M28) in the total digest, allowed purification of these monomers and 24-kDa (M24) and 26-kDa (M26) monomers free from dimers. Reactivity of these subunits with Goodpasture autoantibodies using immunoblotting of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional nonequilibrium pH gradient electrophoresis gels showed strong reactivity with the purified M28 subunits. Inhibition enzyme-linked immunosorbent assay, used to quantitate the reactivity of the purified NC1 subunits, indicated that M28 had a greater than 10-fold increase in ability to inhibit binding to NC1 than NC1 itself. Comparison of hGBM NC1 components were made with those obtained from collagenase digests of salt and acid-extracted bovine and sheep GBM and Englebreth-Holm-Swarm tumor similarly purified by gel filtration and reverse-phase high performance liquid chromatography. Two-dimensional gel analysis of these NC1 isolates revealed absence of the very cationic M28 monomers. Reactivity with antibodies eluted from diseased kidneys of sheep immunized with hGBM (Steblay nephritis) was compared with Goodpasture autoantibody reactivity by immunoblotting two-dimensional gels of hGBM NC1. We conclude that a very cationic M28 monomer (M28 ) found only in hGBM is the probable target in Goodpasture syndrome, that the epitope is present on most NC1 components from extracted and unextracted hGBM, and is exposed by urea denaturation which is enhanced by acid treatment. A weakly cationic M28 monomer (M28+) is present in GBM from other species and is the probable target in Steblay nephritis. Differential recognition of the two M28 components by these antibodies points to different genetic origins or possibly distinct post-translational modifications for these components. This is supported by their presence or absence in different species and tissues, as well as biochemical differences from the M24/26 monomers which presumably are derived from alpha 1(IV) and alpha 2(IV) collagen chains.

Amino Acids↗
PubMed1986

Comparison of non-collagenous type IV collagen components in the human glomerulus and EHS tumor.

A method for the isolation of the NC1 domain of type IV collagen has been developed using the EHS sarcoma, a basement membrane-producing mouse tumor. This NC1 domain has been compared to the NC1 of human glomerular basement membrane (hGBM) to assess its usefulness in the biochemical characterization of the Goodpasture antigen which is associated with NC1. Both NC1 isolates appeared to migrate by gel filtration as hexamers (Mr 160,000) and in SDS-polyacrylamide gel electrophoresis as dimers and monomers (Mr 54,000 and 26,000), and were shown to share biochemical identity by amino acid analysis. The hGBM NC1 showed greater complexity in the monomer region, and when compared by two-dimensional gel electrophoresis was found to contain more components in both regions than EHS NC1. Anti-GBM autoantibodies from patients with Goodpasture's syndrome reacted with the EHS NC1 by immunoblotting of two-dimensional gels. The EHS NC1 isolated by reverse phase HPLC partially inhibited the reactivity of the anti-GBM autoantibodies against hGBM NC1 by inhibition ELISA assay. Reverse phase HPLC elution of EHS and hGBM NC1 showed differences in subunit composition and interaction; complete dissociation of the EHS monomers and dimers in 0.1% trifluoroacetic acid was observed, whereas hGBM monomers and dimers eluted together. Rotary shadowing of hGBM NC1 domains revealed size heterogeneity of globular domains, compared with greater uniformity of EHS NC1 hexamers. We conclude that EHS NC1 contains an epitope(s) that is reactive with human autoantibodies to hGBM NC1. However, the immune response in Goodpasture's syndrome may involve antibodies directed against epitopes which are present in greater density and on a more complex array of peptides in the hGBM NC1 than in EHS NC1.

Amino Acids↗
PubMed1986

Decline of vaccine-induced antipneumococcal antibody in children with nephrotic syndrome.

Twenty-five children with steroid-responsive idiopathic nephrotic syndrome were studied for persistence of antipneumococcal capsular polysaccharide antibody during relapse of their disease and at 1, 6, and 12 months after vaccination with the 14-valent pneumococcal vaccine. Nonrelapsers (group I) were compared to those who had at least one relapse but whose sera were obtained during remission (group II). Group II had a more rapid decline in total anticapsular antibody per month than group I (5.3% v 2.4%). Analysis by individual anticapsular types showed that differences between groups approached significance only for type 4 (P = .07). Rates of decline of antibody against pneumococcal capsular polysaccharide varied among types. One year after vaccination, 50% of patients had less than 300 ngAbN/mL against types 4, 6A, 7F, 8, and 19F. Sera obtained from seven patients during relapse had geometric mean antibody concentrations less than 300 ngAbN/mL against those same types; two of these types have been reported to cause disease in vaccinated patients with nephrosis. Decline of antibody against pneumococcal capsular polysaccharide following vaccination varies by capsular type. Type-specific analysis should be used when monitoring serum antibody levels in these patients after vaccination.

Adolescent↗
PubMed1986

Nephritogenic antigen determinants in epidermal and renal basement membranes of kindreds with Alport-type familial nephritis.

We probed epidermal basement membranes (EBM) of acid-urea denatured skin from members of kindreds with Alport-type familial nephritis (FN) for the presence of antigens reactive with Goodpasture sera (GPS) and serum (FNS) from an Alport patient who developed anti-glomerular basement membrane (GBM) nephritis in a renal allograft. By immunoblotting, GPS reacted primarily with the 28,000 molecular weight (mol wt) monomer but also the 24,000 mol wt and 26,000 mol wt monomers of the noncollagenous globular domain (NC1) of type IV collagen from normal human GBM, while FNS identified only the 26,000-mol wt monomer. FNS reacted with EBM of 12 controls and nine unaffected male kindred members but not EBM of eight affected males. Five affected females exhibited interrupted reactivity of FNS with EBM. GPS showed variable reactivity with EBM and was not discriminating with respect to Alport-type FN. FNS did not stain renal basement members of five affected males. However, the EBM, tubular basement membrane, and Bowman's capsules of affected males contained antigens reactive with GPS. These immunochemical studies suggest that the FNS antigen is distinct from Goodpasture antigen(s). The expression of FNS antigen located on the NC1 domain of type IV collagen is altered in basement membranes of patients with Alport-type FN, and the distribution of this antigenic anomaly within kindreds suggests X-linked dominant transmission of a defective gene.

Basement Membrane↗
PubMed1985

Detection of hidden nephritogenic antigen determinants in human renal and nonrenal basement membranes.

The reactivity of 10 human anti-glomerular basement membrane (GBM) autoantibodies with basement membrane antigens of human adult and infant kidney, lung, placenta, and skin was examined by ELISA and immunofluorescence microscopy. All autoantibodies were previously shown to react with adult kidney by indirect immunofluorescence and with collagenase-digested adult GBM by ELISA. Four antibodies (group A) were positive on infant and fetal kidney sections by immunofluorescence, and six antibodies were negative (group B). By ELISA both groups of antibodies were reactive with collagenase digests of infant GBM. After denaturation with 6M urea (pH 3.5) infant and fetal kidney sections reacted with group B autoantibodies by immunofluorescence, which indicated that the antigen(s) was masked. Hidden antigenic determinants in lung, placenta, and skin were reactive with groups A and B autoantibodies only after acid urea denaturation of tissue sections. Within each group variability in reactivity of autoantibodies with basement membranes suggested further heterogeneity.

Adult↗
PubMed1985

Analysis of nephritogenic antigens in human glomerular basement membrane by two-dimensional gel electrophoresis.

Collagenase digests of GBM were partially purified by column chromatography and analyzed by 2-D gel electrophoresis. Silver staining of 2-D gels showed charge- and size-related heterogeneity of proteins in the 45 to 50 kDa and 25 to 27 kDa regions. These components were transferred to nitrocellulose sheets and reacted with 10 human anti-GBM autoantibodies. Detection of bound anti-GBM autoantibodies to blotted proteins was carried out with peroxidase-labeled goat anti-human IgG and revealed binding predominantly to the cationic (pI 8 to 9.0) 45 to 50 kDa and 25 to 27 kDa components. Positive-staining patterns of blotted proteins were similar with all anti-GBM autoantibodies except that three sera additionally identified neutral (pI 5.5 to 6.5) protein components. One anti-GBM autoantibody, which developed following renal transplantation, lacked reactivity with the most cationic components in the 25 to 27 kDa region. These findings suggest heterogeneity of nephritogenic GBM antigens. The cationic 45 to 50 kDa components were sensitive to reduction, while one neutral 45 to 50 kDa component was resistant; a complex array of 25 to 30 kDa proteins (pI 5.5 to 7.5) were observed by silver staining postreduction. None of the reduced protein components reacted with anti-GBM antibodies, suggesting that epitopes on nephritogenic GBM antigens may be related to disulfide-bonded regions. Although there is variable immunohistochemical reactivity of anti-GBM autoantibodies with the GBM of infant kidneys, 2-D gels of collagenase-digested human infant GBM blotted and reacted with anti-GBM autoantibodies and showed staining patterns similar to that of adult GBM. These studies demonstrate the presence of nephritogenic antigens in the GBM of immature human kidney which are not detectable by immunohistochemical analysis.

Adult↗
PubMed1985

Enzyme immunoassay of anti-glomerular basement membrane antibodies.

An immunoassay has been developed to detect anti-glomerular basement membrane (GBM) antibodies in human sera. Various plating conditions, types of microtiter plates, and the use of biotinylated or peroxidase-labeled secondary antibodies were examined. The described assay is reliable, fast, and convenient. Sera with positive reactivity in anti-GBM nephritis and Goodpasture's syndrome are readily discriminated from sera obtained from normal individuals or patients with a variety of other diseases.

Anti-Glomerular Basement Membrane Disease↗
PubMed1984

Detection of Goodpasture antigen in fractions prepared from collagenase digests of human glomerular basement membrane.

Preparations of human glomerular basement membrane (GBM) were digested with collagenase, and a Goodpasture (GP) antigen rich pool from gel filtration column runs was identified by antibody inhibition radioimmunoassay. The components of the GP antigen pool were separated on polyacrylamide gels, and transferred to nitrocellulose sheets by the 'western' blotting technique. The blots were separately reacted with thirteen GP sera as primary antibody, followed by peroxidase labelled goat anti-human IgG and revealed 45-50K (two bands) and 25-28K (one-three bands) components. No corresponding reactivity was observed using convalescent GP sera or other control sera (normal human serum, rapidly progressive glomerulonephritis with or without pulmonary haemorrhage, and lupus erythematosus) as primary antibody.

Amino Acids↗
PubMed1984

Development and heterogeneity of antigens in the immature nephron. Reactivity with human antiglomerular basement membrane autoantibodies.

Indirect immunofluorescence microscopy was performed with 15 human anti-glomerular basement membrane (GBM) antibodies and mouse monoclonal antibodies to Type IV collagen (MBM4) and renal basement membranes (MBM15) on renal tissue from 6 fetuses (gestational age, 15-23 weeks), 8 infants (age, 1-21 days), and 8 children and adults (ages, 3-27 years). Of the 15 human anti-GBM antibodies that react with GBM in adult glomeruli, only 4 identified antigens in the GBM of fetal and infant glomeruli. In contrast, the monoclonal antibodies bound to basement membranes in the uninduced nephron and the GBM throughout all development stages of the fetal kidney. These studies demonstrate that the reactivity of human autoantibodies with GBM is developmentally and gestationally related--some identifying an antigen(s) in fetal glomeruli with early capillary loop formation and others reacting only with GBM in fully mature kidneys.

Adolescent↗
PubMed1983

Absence of Goodpasture's antigen in male patients with familial nephritis.

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.

Anti-Glomerular Basement Membrane Disease↗
PubMed1982

Serum antibody response to pneumococcal vaccine in children with nephrotic syndrome.

Children with nephrotic syndrome are susceptible to serious pneumococcal disease and may be immunodeficient on the basis of abnormal humoral immune responses to natural antigens or immunoglobulin loss during relapse. As part of an ongoing study to evaluate pneumococcal anticapsular antibody concentration and immunologic competence, 27 steroid-responsive and six steroid-resistant patients with nephrotic syndrome, and 12 age-matched control subjects, were vaccinated with polyvalent pneumococcal vaccine. Antibody responders were defined as patients with at least a twofold increase in antibody after vaccination as well as an antibody concentration greater than 200 ng of anticapsular antibody nitrogen per milliliter (ngN/ml) after vaccination. Pneumococcal antibody concentrations before and after vaccination were significantly depressed in steroid-resistant patients when compared with control subjects (P less than .002) and with the steroid-responsive nephrotic syndrome group (P less then .001). Steroid-responsive nephrotic children who were not receiving corticosteroid therapy at the time of vaccination had significantly higher antibody concentrations to five pneumococcal types before vaccination and to seven types after vaccination compared with control subjects (P less than .05). Fewer steroid-responsive patients receiving corticosteroids achieved antibody concentrations greater than or equal to 200 ngN/ml against type 19F compared with patients not receiving steroids or with control subjects (P less than .05). These results suggest that pneumococcal vaccine is immunogenic in children with steroid-responsive nephrotic syndrome and may protect these patients from disease due to pneumococcal types contained in the vaccine.

Adolescent↗
PubMed1982

Systemic lupus erythematosus in the first two decades of life.

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.

Adolescent↗
PubMed1981

Distribution of glomerular basement membrane antigens in diseased human kidneys.

We have previously shown, using high resolution epifluorescence microscopy, that heterologous rabbit antihuman glomerular basement membrane (GBM) antibody (RAHGBM) stains the GBM of normal human kidney in a bilaminar distribution along the endothelial and epithelial aspects of the GBM. In contrast, Goodpasture (GP) antibody stains a single linear component along the inner aspect of the GBM. In the present study, alterations in the staining reactivity of these components appear consistently to correlate with the known morphologic changes in various glomerulopathies. In idiopathic nephrotic syndrome, no changes from normal were observed. In membranous glomerulopathy, the inner RAHGBM antigen was preserved, but the outer antigen showed an irregular beaded alteration adjacent to membranous GBM immune deposits. In diabetic glomerulopathy, striking increase in the inner RAHGBM component was evident with markedly increased separation from the outer RAHGBM line. In membranoproliferative glomerulonephritis (MPGN), loss of the inner component but preservation of the outer RAHGBM line was evident; however, in type I MPGN, GP antigen was preserved as a single linear component, but in type II MPGN, there was splitting of GP antigen by GBM-dense deposit material into a double linear array along the endothelial and epithelial aspects of the GBM. These observations serve to substantiate our earlier findings that antigenic and nonantigenic components of the GBM are partitioned in the glomerular capillary wall; their selective characteristic alteration in various glomerulopathies additionally supports this view.

Antigens↗
PubMed1981

Renal failure as a complication of acute antihypertensive therapy.

Adult patients with long-standing hypertension have been reported to experience an impairment in renal function when treatment with potent vasodilating agents is initiated. To document that this sequence may occur in children as well, we report the case of a 4-year-old boy with renal disease in whom reduction of blood pressure to normal levels was accompanied on three occasions by oliguric renal failure. During each episode, the correlation between reduction in blood pressure and increase in serum creatinine level was significant (P less than .05); furthermore, the slope of the relationship was similar with each episode. This phenomenon suggests an impairment of renal autoregulation in this patient. Maintenance of normal blood pressure for several months was accompanied by a gradual return of renal function to pretreatment levels. This case suggests that particular attention should be paid to renal function during the initiation of antihypertensive therapy, particularly in patients with renal vascular damage. Present evidence does not appear to warrant modification of the current therapeutic philosophy of aggressive management in patients with severe hypertension.

Acute Kidney Injury↗