Murine Autoimmune Interstitial Nephritis and Associated Antigen: Purification of a Soluble Tubular Basement Membrane Antigen from Mice Kidneys

Abstract

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 of interstitial nephritis in mice produced with a purified TBM antigen.

Effects of Bupleurum scorzoneraefolium, Bupleurum falcatum, and saponins on nephrotoxic serum nephritis in mice

“…250 mesh was disrupted by sonication to separate the cellular components from the GBM. An adequate amount of trypsin (25 mg trypsin/10 g GBM) was added to isolate the GBM-rich fraction in 0.1 M Tris-HCl buffer containing 0.02 M CaCl 2 (pH 8.2), and this mixture was incubated while stirring at 37 • C for 18 h. After centrifugation at 4 • C and 10,000 rpm for 60 min, a supernatant containing the crude solubilized GBM antigen (GBM-Ag) was obtained (Wakashin et al, 1980).…”

Infusion of mesenchymal stem cells overexpressing GDNF ameliorates renal function in nephrotoxic serum nephritis

“…Three rabbits were subcutaneously immunized with 4 mg of GBM antigen four times at 10‐day intervals. Antisera collected from the animals 7 days after the final antigen injection were pooled and used as NTS throughout the study . Five days later, rats housed in cages with a high‐efficiency particulate air filter and fed a standard rat diet were intravenously injected with 200 µl of NTS.…”

“…Antisera collected from the animals 7 days after the final antigen injection were pooled and used as NTS throughout the study. 13 Five days later, rats housed in cages with a highefficiency particulate air filter and fed a standard rat diet were intravenously injected with 200 ml of NTS. Control rats were injected with 200 ml of normal rabbit serum.…”