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Biomedical subjects

M Wakashin

Publications and source records attributed to M Wakashin.

53 records · Page 3Linked to original sources

Murine autoimmune interstitial nephritis and associated antigen: purification of a soluble tubular basement membrane antigen from mice kidneys.

A soluble tubular basement membrane (TBM) antigen has been purified from murine kidneys. The isolated fibers were trypsinized and subjected to zone electrophoresis, Sephadex G-200 gel filtration, and DEAE-cellulose chromatography eluted with linear NaCl concentration gradient while monitoring the TBM antigen activity with specific antihuman TBM antigen. The molecular weight of TBM antigen was estimated to be 30,000 daltons by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified TBM antigen gave a single precipitin line against the specific antiserum and cross-reacted with the TBM antigen similarly purified from other animals such as goat, guinea pig and also with human TBM antigen in an immunodiffusion plate. BALB/c, C3H/He and C57BL/6 mice were immunized twice at 2 weeks' interval with the TBM antigen and adjuvant. Histologically typical interstitial nephritis occurred in BALB/c mice, whereas no nephritis developed in other strains of mice. The inflammatory changes were characterized by intense mononuclear cell infiltration, tubular destruction, and interstitial and periglomerular fibrosis. The serum-stained basement membrane of normal tubules and Bowman's capsules, and no antiglomerular basement membrane activity was detectable in the serum by immunofluorescence. This system provides a simple and useful model in interstitial nephritis in mice produced with a purified TBM antigen.

Animals↗

Association of gastric cancer and nephrotic syndrome. An immunologic study in three patients.

Three patients presenting typical nephrotic syndrome concurrent with gastric cancer are described. The carcinoma was moderately advanced and was histologically an adenocarcinoma in all cases. Morphologic examination of the renal biopsy revealed the characteristic changes of membranous nephropathy and typical subepithelial electron-dense deposits. The glomerular deposits were specifically stained with antihuman IgG and beta-1-c. The eluate of the glomeruli reacted specifically with the surface of the cancer cells from the same patient when studied by the immunofluorescent method. The eluate also reacted with the cancer cell extract as examined with the radioimmunodiffusion technique and with carcinoembryonic antigen (CEA) prepared by the method of Gold. Furthermore, the nephrotic syndrome completely remitted in 1 patient following surgical removal of the cancer. These bits of evidence indicate a possible role of a stomach cancer related antigen and/or CEA in the development of immune complex nephropathy in these patients.

Adenocarcinoma↗

Immunochemical characterization and quantitation of the human glomerular basement membrane antigen from the urine of patients with glomerular diseases.

A soluble glomerular basement membrane (GBM) antigen was detected in the urines of patients with various glomerular diseases including chronic glomerulonephritis, nephrotic syndrome, chronic renal insufficiency, and lupus nephropathy. The urinary GBM antigen (u-GBM) was immunochemically distinct from other renal antigens and other serum components, but it was cross-reactive with trypsinized human GBM antigen (t-GBM). The molecular size of u-GBM was approximately the same as human serum albumin as estimated by elution patterns on Sephadex G-200. The concentration of u-GBM was estimated quantitatively by a single radial radioimmunodiffusion. Although differed from case to case, a rough correlation with the type and/or stage of nephrotic syndrom existed. It was also demonstrated that the amounts of u-GBM decreased in response to steroid therapy of nephrotic syndrome. It was further shown that in a case of membranoproliferative glomerulonephritis, anti-GBM antibody could be eluted from the kidney removed from the patient. These findings imply that the GBM antigen plays an important role in the pathogenesis of human renal diseases. The pathophysiological significance of urinary GBM excretion in renal diseases is also discussed here.

Antigens↗

Renal tubular dysfunction in Minamata disease. Detection of renal tubular antigen and beta-2-microglobin in the urine.

"Minamata disease" was found among the residents along Minamata bay contaminated with the effluent from an industrial plant using mercury. The patients were suffering from various neurologic disorders primarily due to organic mercury poisoning. Evidence is described of renal tubular dysfunction associated with this disease by the immunochemical demonstration or renal tubular epithelial antigen and beta-2-microglobulin in the urine. Nineteen patients with Minamata disease and 35 diseased and healthy control subjects were examined. The contents of urinary renal tubular epithelial antigen and beta-2-microglobulin, and the ratios of these proteins to albumin in individuals with Minamata disease were significantly different from the levels in healthy control subjects (P less than 0.05) were identical to those found in patients with tubular and the values, proteinuria. These results indicate that Minamata disease is associated with renal tubular dysfunction, and also suggest that these procedures would be useful for screening the nephrotoxicity in the environmental exposure of heavy metals.

Adult↗

[Urinary proteins].

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Chromatography, Gel↗

Cell-mediated cytotoxicity of sensitized spleen cells against target liver cells--in vivo and in vitro study with a mouse model of experimental autoimmune hepatitis.

Spleen cells obtained from C57BL/6 (B6) mice with an experimental autoimmune hepatitis were transferred to normal C57BL/6 recipient mice. Most prominent liver damages occurred in the recipient mice injected with sensitized nylon wool column-adherent spleen cells from the donor mice. Production of such liver damage was blocked by treatment of the sensitized adherent spleen cells with anti-Thy 1,2 monoclonal antibody and complement before injection. Based on these in vivo results, a microcytotoxicity assay was performed using isolated C57BL/6 hepatocytes as target cells and sensitized spleen cells obtained from hepatitis donor mice as effector cells. The fraction of sensitized nylon wool-adherent spleen cells demonstrated a high cytotoxic activity against isolated syngeneic hepatocytes, although the other fractions and spleen cells of control animals showed no such effect. The cytotoxic activity of sensitized-adherent spleen cells against target hepatocytes was significantly reduced after treatment with anti-Thy 1,2 antibody and complement, but it increased after depletion of B cells and Fc receptor-bearing T-cells. Although these sensitized nylon wool-adherent spleen cells showed high cytotoxic activities against syngeneic hepatocytes, their cytotoxicity against allogeneic hepatocytes was lower. They exerted no cytotoxic activity against syngeneic renal glomerular cells and EL-4 thymoma cells. These results suggest that sensitized T-cells in the nylon wool column-adherent fraction play the role of cytotoxic killer cells against target liver cells in vitro.

Animals↗

Effect of glycyrrhizin on lysis of hepatocyte membranes induced by anti-liver cell membrane antibody.

Studies were made on why glycyrrhizin injection decreases the plasma aspartate aminotransferase (AST) and alanine aminotransferase activities in patients with chronic hepatitis. For this, rat hepatocytes were isolated, and incubated with antibody raised against rat liver cell membranes, and the effect of glycyrrhizin on their release of transaminase was investigated. Isolated rat hepatocytes released AST on incubation with anti-liver cell antibody in the presence of complement. At this time, their endogenous phospholipase A2 activity was increased. Cultured hepatocytes also released the transaminase in the presence of venom phospholipase A2. Glycyrrhizin suppressed the release of transaminase in the presence of either anti-liver cell membrane antibody or phospholipase A2. These results suggest that antibody treatment raised the phospholipase A2 activity in liver cell membranes, resulting in release of transaminases, and that glycyrrhizin suppressed this increase in phospholipase A2 activity and so inhibited the release of transaminase.

Animals↗