Proteinuria and structural alterations in rat glomerular basement membranes induced by intravenously injected anti-laminin immunoglobulin G.

Abstract

Antibodies against laminin, which is a defined glycoprotein of basement membranes, were produced in sheep and affinity purified by immunoadsorption on laminin-Sepharose (S alpha L). When injected intravenously into rats, S alpha L rapidly bound in a linear pattern to the glomerular basement membrane (GBM) in the peripheral and mesangial regions of all glomeruli, and, when greater than 0.5 mg S alpha L was injected, to some tubular BM as well. 1-2 h after the injection of conjugates of horseradish peroxidase (HRP) and S alpha L, HRP reaction product was present throughout the full thickness of the GBM and mesangial matrix. [125I]S alpha L binding to the kidney in vivo increased linearly over the dose range of 40-950 micrograms of IgG and accounted for approximately 2% of the injected dose/g kidney. When 4 mg of [125I]S alpha L was injected, 1.5% or 62 micrograms/g kidney was bound. Proteinuria did not develop within 7 wk of injection in rats that received 0.5-1.6 mg of S alpha L. In contrast, all animals that received injections of 4 mg of S alpha L gradually became proteinuric within 3-6 wk. Thickening, reduplication, and flocculent subendothelial deposits were observed in the GBM of these animals. In addition, mononuclear cells adhered to the GBM and infiltrated beneath the endothelium. However, the deposition of rat C3 was infrequently observed, and rat IgG was not seen in the glomeruli of any rat that received S alpha L. 10 wk after injection, much greater amounts of S alpha L appeared within the mesangium than the peripheral GBM. These results demonstrate that the interaction of S alpha L with the GBM, possibly in concert with infiltrating mononuclear cells, gradually altered the structure and permeability characteristics of the glomerulus independent of a host anti-S alpha L humoral response.

Epithelial basement membrane of mouse jejunum. Evidence for laminin turnover along the entire crypt-villus axis.

“…The presence ofother extracellular matrix components in this laminin preparation, including fibronectin, proteoglycans, or entactin/nidogen was not detected immunochemically (25), on polyacrylamide gels (24,25,27) or by rotary shadow electron microscopy (27). Anti-laminin IgG from previously immunized sheep was affinity-purified as described earlier (24,25,28) using laminin-Sepharose columns and was highly specific for laminin with no cross-reactivity with type IV collagen by an enzyme-labeled immunosorbent assay (28) or by Western blotting (data not shown). Sheep anti-laminin IgG and, for control studies, chromatographically pure sheep IgG (Organon Teknika-Cappel Laboratories, West Chester, PA) were conjugated directly (29) to activated HRP (type VI, Sigma Chemical Co., St. Louis, MO) as before (24,25,28).…”

“…Laminin from murine Englebreth-HolmSwarm tumor was purified (25,26). The presence ofother extracellular matrix components in this laminin preparation, including fibronectin, proteoglycans, or entactin/nidogen was not detected immunochemically (25), on polyacrylamide gels (24,25,27) or by rotary shadow electron microscopy (27).…”

“…Anti-laminin IgG from previously immunized sheep was affinity-purified as described earlier (24,25,28) using laminin-Sepharose columns and was highly specific for laminin with no cross-reactivity with type IV collagen by an enzyme-labeled immunosorbent assay (28) or by Western blotting (data not shown). Sheep anti-laminin IgG and, for control studies, chromatographically pure sheep IgG (Organon Teknika-Cappel Laboratories, West Chester, PA) were conjugated directly (29) to activated HRP (type VI, Sigma Chemical Co., St. Louis, MO) as before (24,25,28). Rhodamine-conjugated rabbit anti-sheep IgG was obtained from Organon TeknikaCappel Laboratories.…”

“…Laminin from murine Englebreth-HolmSwarm tumor was purified (25,26). The presence ofother extracellular matrix components in this laminin preparation, including fibronectin, proteoglycans, or entactin/nidogen was not detected immunochemically (25), on polyacrylamide gels (24,25,27) or by rotary shadow electron microscopy (27). Anti-laminin IgG from previously immunized sheep was affinity-purified as described earlier (24,25,28) using laminin-Sepharose columns and was highly specific for laminin with no cross-reactivity with type IV collagen by an enzyme-labeled immunosorbent assay (28) or by Western blotting (data not shown).…”

Severe necrotizing pancreatitis following combined hepatitis A and B vaccination

“…10 Vaccines have been shown to induce the production of autoantibodies in several animal models. [11][12][13] In this theory, immunologic injury may be caused by a cytotoxic antibody system that has a heterophilic reactivity to acinar cells. 9 There are also suggestions in the literature of several additional mechanisms of immunologic injury such as polyclonal activation (adjuvant reaction) of lymphocytes, "bystander activation" of self-reactive lymphocytes or somatic mutation of immunoglobulin variable genes.…”