Basement Membrane Preparation Research Articles

Editorial: Anti-tubular basement-membrane (TBM) nephritis.

Despite a distinctive immunofluorescence picture, anti-TBM antibody accumulation in human renal disease was recorded only after descriptions of experimental models of anti-TBM disease. Immunization of guinea pigs or certain strains of rats with appropriate renal tubular basement-membrane (TBM) preparations in adjuvant results in severe renal disease, characterized by tubular damage and interstitial inflammation.1 2 3 The immunofluorescence findings are highly characteristic: completely continuous (linear) accumulation of IgG, and usually of C3, is seen along the TBM (both proximal and distal). (Immunization of rabbits or rats with soluble extracts of kidney or exogenous antigens may also lead to immune-complex deposition along the TBM; . . .

Enzymes in lacrimal secretion

1.1. Pairs of glomerular and tubular basement membrane preparations were isolated from kidney cortex of cattle, pig, sheep, horse, man and rat. Tubular basement membranes were isolated from rabbit kidney cortex.2.2. Purity of preparations was established with light- and electron microscopy and by estimating total phosphorus content.3.3. Marked species dependent variations were found for the following components and ratios: with glomerular basement membranes for the contents of aspartic acid, threonine, serine, glycine, tyrosine and hexosamines; with tubular basement membranes for the contents of alanine, isoleucine, fucose and mannose; with both types of basement membranes for the contents of histidine, glucose and galactose and the ratios of hydroxylated to nonhydroxylated proline and lysine residues, but not for the ratio of 4-hydroxyproline to hydroxylysine.4.4. Comparison of glomerular and tubular basement membranes as pairs from individuals shows differences in all species. As a common phenomenon, the glomerular basement membrane preparations of the six species contain about twice as much neuraminic acids and more mannose and hexosamines than their tubular counterparts.5.5. Observed differences may be related to variations in antigenic properties of the different types of renal basement membranes.

Insulin-binding impairment of plasma membrane receptors in heriditary diabetic mice

1.1. Pairs of glomerular and tubular basement membrane preparations were isolated from kidney cortex of cattle, pig, sheep, horse, man and rat. Tubular basement membranes were isolated from rabbit kidney cortex.2.2. Purity of preparations was established with light- and electron microscopy and by estimating total phosphorus content.3.3. Marked species dependent variations were found for the following components and ratios: with glomerular basement membranes for the contents of aspartic acid, threonine, serine, glycine, tyrosine and hexosamines; with tubular basement membranes for the contents of alanine, isoleucine, fucose and mannose; with both types of basement membranes for the contents of histidine, glucose and galactose and the ratios of hydroxylated to nonhydroxylated proline and lysine residues, but not for the ratio of 4-hydroxyproline to hydroxylysine.4.4. Comparison of glomerular and tubular basement membranes as pairs from individuals shows differences in all species. As a common phenomenon, the glomerular basement membrane preparations of the six species contain about twice as much neuraminic acids and more mannose and hexosamines than their tubular counterparts.5.5. Observed differences may be related to variations in antigenic properties of the different types of renal basement membranes.

Studies on the nature and significance of connective tissue antibodies in adult coeliac disease and Crohn's disease.

Sera from 92 patients with adult coeliac disease, 20 patients with Crohn's disease, and 89 control patients were tested for the presence and immunoglobulin class of anticonnective tissue antibodies. In adult coeliac disease patients taking a normal diet there was a high frequency of sera positive for connective tissue antibodies and the majority were IgA in class. Following gluten withdrawal, IgA class connective tissue antibody activity was strikingly reduced. No such antibody activity was detected in concentrated jejunal fluid collected from 15 patients with adult coeliac disease, seven of whose sera were positive for connective tissue antibody. Connective tissue antibody activity was not absorbed by fractions of gluten. Similarly, preparations of human reticulin and glomerular basement membrane together with elastin and collagen failed to absorb connective tissue antibody.

Specificity of Autoantibodies to Tubular and Glomerular Basement Membranes Induced in Guinea Pigs

Abstract All twenty-eight guinea pigs immunized with a variety of bovine renal basement membrane (RBM) preparations developed similar interstitial nephritides, with minimal glomerular abnormalities. Direct immunofluorescence revealed linear deposition of IgG along the tubular basement membranes (TBM) of all guinea pigs and along the glomerular basement membranes (GBM) of 13 of them. Elution of renal-bound IgG demonstrated that all guinea pigs had antibodies against both TBM and GBM, although anti-GBM activity was usually not detectable at low concentrations of IgG in the eluates. Absorption of pooled eluates with a variety of guinea pig RBM immunoabsorbents revealed the following: 1) approximately three-fourths of the eluted IgG comprised antibody to RBM; 2) all of the eluted autoantibody reacted with TBM, while only 36 to 46% crossreacted with GBM; 3) TBM appeared to have antigenicities both separate from and shared with GBM; 4) guinea pigs immunized with whole bovine TBM appeared to form antibodies to both the collagenous and noncollagenous portions of RBM.

Action of snake venom hemorrhagic principles on isolated glomerular basement membrane

1. 1. The reaction of venom hemorrhagic principles (HR1, HR2a and HR2b) with isolated glomerular basement membrane was studied in vitro. For comparison, experiments were also done with a bacterial collagenase which is known to induce the hemorrhage. Upon incubation of the basement membrane preparation with either of the four agents, its supernatant fluid gave positive reactions for proteins (or peptides) and carbohydrates. The optimal pH of the protein-liberating activities of HR1, HR2a and HR2b was around 8.0. 2. 2. The approximate ratio of the proteins or peptides liberated by HR2a, HR2b, HR1 and collagenase, under standard conditions, was 1:1:2:4; and for the carbohydrates liberated 0.5:0.5:2:4, thus suggesting that HR2a or HR2b acted on the basement membrane in a different manner from HR1. This interpretation is supported by comparison between the split-products of the membrane by these principles in gel filtration experiments. 3. 3. Liberation of both proteins and carbohydrates from basement membrane by HR1, HR2a and HR2b was inhibited by EDTA, cysteine, antivenin and antihemorrhagic factor from snake serum, all of which inhibit the hemorrhagic activity of these principles. The results suggest that one and the same entity is responsible for both the hemorrhagic and liberating activities in each venom principle. 4. 4. Purified HR1 was resolved partially into four components by polyacrylamide gel electrophoresis. Both liberating and hemorrhagic potencies, as expressed by their specific activities, however, were constant throughout, which further supports the idea that the same factor is responsible for both activities. 5. 5. The hemorrhagic effect of HR1, HR2a, HR2b and collagenase is attributed to enzymatic destruction of the basement membrane with consequent lowering of the stability of the vessel wall.

Collagen crosslinks: Occurrence in basement membrane collagens

Basement membrane preparations from anterior lens capsule, Descemet's membrane, and renal glomerulus were reduced with NaB 3H 4 in order to label carbonyl-derived crosslinks. Quantitative incorporation of tritium into the basement membranes was found, similar to the levels observed in fibrous collagens. A considerable proportion of the label was chromatographically identical with the reduced aldehydes and crosslinks of collagen, suggesting that is is the collagenous portions of basement membrane which contain these compounds. This was substantiated by showing that the isolated collagenous proteins contained substantial amounts of reduced aldehydes and crosslinks.

Phospholipid of the rat glomerular basement membrane in experimental nephrosis.

Phospholipids were found to be a constant component of rat glomerular basement-membrane preparations. The concentration fell during preparation of basement membrane by sonication of whole glomeruli, but then remained constant despite continued sonication. The proportions of the individual phospholipids were different from those of whole renal tissue or of isolated glomeruli. The basement-membrane preparations had no (Na(+)+K(+))-activated adenosine triphosphatase activity, an enzyme that is bound to plasma membranes. The concentration of lipid P was decreased on exposure in vivo or in vitro to antiserum against basement membrane; 7 days after injection of antiserum there was a change in the phospholipid composition, with a relative increase in phosphatidylcholine and a decrease in sphingomyelin content. The metabolic turnover rate of the lipid P remaining in the membrane was normal, as determined by (32)P incorporation. The loss of phospholipid was associated with decreases in the relative concentrations of hydroxyproline, hydroxylysine and glycine, and relative increases in proline, lysine, serine, threonine and valine. Administration of aminonucleoside and daunomycin produced proteinuria but did not cause a decrease in lipid P. Anticollagen and anti-lymphocyte sera that attached to the basement membrane but failed to produce proteinuria, also failed to affect the phospholipid content.

Variations in the sialic acid concentration of glomerular basement membrane preparations obtained by ultrasonic treatment.

Several authors (10, 15, 6, 1, 20) have suggested that sialic acid is a component of the glomerular basement membrane (BM) . Investigations by Spiro (20) have shown that BM preparations obtained by ultrasonic treatment of whole glomeruli contain two distinct carbohydrate moieties, in approximately equal proportions : one is a disaccharide containing glucose and galactose, believed to be intrinsically associated with collagen, and the other is a heteropolysaccharide consisting of galactose, mannose, hexosamines, sialic acid, and fucose . We have presented histochemical evidence that sialic acid is present in the glomerulus and that it is located not in the basement membrane proper but in the cell membranes lining the basement membrane (18, 14). Preceding us, Rambourg and Leblond (17) demonstrated a carbohydrate-rich cell coat, acidic in nature, at the surface of glomerular cells, and recently Gronowski et al . (3) and Jones (4) published results similar to ours . The present study was undertaken in an attempt to demonstrate that basement membrane preparations obtained by ultrasonic treatment, a customary procedure for biochemical analysis, are contaminated by cell membranes and that the sialic acid content of such preparations increases with the rising centrifugal force at which they are collected .

Chemical structure of tubular and glomerular basement membranes of human kidney. Isolation, purification, carbohydrate and amino acid composition.

A method involving the use of sieves and differential centrifugation for the preparation and separation of basement membranes from human renal tubules and glomeruli is described. Electron microscopic examination of the material obtained shows the glomerular membrane as a single dense structure, while the tubular membrane is composed of two dense layers separated by a space filled with loose fibrils exhibiting the same periodicity as collagen. The carbohydrate composition of both types of membranes is the same but tubular membranes have a lower content of some specific sugars. Differences also exist in amino acid composition. The content in hydroxylysine is higher in the glomerular membrane while the content in hydroxyproline is higher in the tubular membrane.Gel electrophoresis after solubilization of the material demonstrates the presence of two, may be three, major proteins.

Effect of injecting heterologous glomerular basement membrane preparations in pregnant sheep.

Summary(1) Three sheep developed a fatal nephritis in 45 to 54 days when immunized with heterologous GBM and Freund's adjuvant in the last third of their pregnancy. (2) Immunization of the mothers or suckling their colustrum (in 2 of the 4 lambs) did not produce a renal lesion in the lambs. (3) It appears that the agent which produces a fatal nephritis in the mother was either not transmitted in utero or in the colustrum or it was incapable of affecting the fetal or neonatal kidney. (4) The above observations are consistent with the hypothesis that this sheep nephritis is produced by an autoimmune mechanism mediated by sensitized cells and/or autoantibody.

Some Immunologic Properties of Human and Dog Glomerular Basement Membranes

Summary A consistently useful technique for producing human glomeruli and human glomerular basement membrane preparations has been demonstrated, based on the Krakower-Greenspon method. One human glomerular preparation was microscopically free of any capsular or tubular contaminants. Human and dog glomerular basement membrane preparations were found to produce CF antibodies in the rabbit. An in vitro cross reaction between human and dog glomerular basement membrane preparations was demonstrated by means of a CF test. The homologous titer was greater in all cases. The supernatant of the low speed centrifugate of the oscillated glomeruli, largely cellular, was found to be a good antigen in the in vitro CF test. It is not clear whether this is due to solubilized and fragmented basement membrane or shorn away cellular cytoplasm antigens or both. The most likely explanation for the above facts is that immunologically similar if not identical antigen(s) exist in dog and human glomerular basement membrane preparations of the purity herein described.

Secondary rejection phenomenon elicited by primary homograft in pretreated rats.

SummaryIntraperitoneal injections of pooled rat glomerular basement membrane preparations into rats subsequently homografted with a primary orthotopic skin graft facilitated a secondary rejection phenomenon on the part of such treated animals. The low nucleic acid content of the preparations limited correlation with the hypothesis of nucleo-proteins as the transplantation antigens.