ELECTRON MICROSCOPIC STUDIES OF EXPERIMENTAL NEPHRITIS WITH FERRITIN-CONJUGATED ANTIBODY

Acute, subacute, and chronic glomerulonephritis, similar in certain features to human glomerulonephritis, has been produced in rabbits by repeated injections of bovine serum albumin. The ratio of antigen to antibody was the factor determining the development and type of glomerulonephritis. This is in confirmation of the observations of Dixon, Feldman, and Vazquez. With the aid of the ferritin antibody technique it was shown that antigen aggregates (probably antigen-antibody complexes) are present in the blood, cross the endothelium and the basement membrane, and accumulate as dense deposits between the basement membrane and the epithelial cytoplasm. In the deposits electron-dense aggregates formed by antigen or by antigen-antibody complexes and material which might be other endogenous proteins may be identified. In rabbits dead of anaphylactic shock following injection of bovine serum albumin, dense material was found within glomerular capillaries, presumably formed by the embolic deposition of antigen-antibody complexes, since the immunofluorescein and immunoferritin techniques demonstrated the presence of both BSA and rabbit globulin.

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Albumin reduces basement membrane hydraulic conductance in part due to arginyl side groups

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1995.269.5.h1514 ◽

1995 ◽

Vol 269 (5) ◽

pp. H1514-H1521 ◽

Cited By ~ 1

Author(s):

M. A. Katz ◽

M. L. La Marche

Keyword(s):

Extracellular Matrix ◽

Bovine Serum Albumin ◽

Basement Membrane ◽

Serum Albumin ◽

Protective Effect ◽

Binding Sites ◽

Bovine Serum ◽

Hydraulic Conductance ◽

Experimental Control ◽

Low Concentrations

Albumin reduces capillary hydraulic conductance (Lp) even at low concentrations. To determine if part of this barrier protective effect might be extracellular, we studied the effects of bovine serum albumin (BSA) on Lp of self-assembled basement membrane (Matrigel). Lp with tris(hydroxymethyl)aminomethane (Tris) buffer superfusate was stable at 1.77 +/- 0.22 x 10(-5) (SE) cm.s-1.cmH2O-1 over several hours. At 0.1 g/dl BSA, experimental/control (Tris) Lp fell to 83.1 +/- 6.0% (2P < 0.025), with decreases to 72.4 +/- 3.7% at 1 g/dl (2P < 0.005), 45.3 +/- 5.1% at 2.5 g/dl (2P < 0.001), and 45.0 +/- 4.8% at 4.0 g/dl (2P < 0.001). In separate experiments, BSA arginine groups were neutralized by 1,2-cyclohexanedione (CHD), and experimental/control Lp values were measured. At 2.5 g/dl, CHD-BSA depressed Lp to 54.4 +/- 4.8%, while unmodified BSA reduced Lp to 40.8 +/- 3.5% of Tris control (2P = 0.05). Finally, soluble arginine at three- and sixfold the arginine in BSA was added to BSA superfusate. For threefold, Lp rose to 120 +/- 8% of BSA level and for sixfold to 129 +/- 9% (2P < 0.05). We conclude that some part of the albumin protective effect is very likely due to consequences on extracellular matrix and that at least 18-22% of this effect is related to arginine groups on albumin when computed from Lp, and up to 34% when viscosity is taken into account. Membrane-saturable arginine-binding sites can be unbound with arginine, thus nullifying part of the barrier protective effect of BSA.

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Clogging of the glomerular basement membrane.

The Journal of Cell Biology ◽

10.1083/jcb.93.2.489 ◽

1982 ◽

Vol 93 (2) ◽

pp. 489-494 ◽

Cited By ~ 70

Author(s):

Y S Kanwar ◽

L J Rosenzweig

Keyword(s):

Glomerular Filtration Rate ◽

Bovine Serum Albumin ◽

Basement Membrane ◽

Serum Albumin ◽

Glomerular Filtration ◽

Glomerular Basement Membrane ◽

Bovine Serum ◽

Filtration Rate ◽

Inulin Clearance ◽

Selective Filter

The negative charges of the sulfated glycosaminoglycans (GAGs) of the glomerular basement membrane (GBM) were differentially neutralized by perfusin with high molarity buffers in order to determine whether or not these charges protect the GBM from being clogged by circulating plasma macromolecules. Progressive elimination of the negative charges resulted in clogging of the GBM by perfused native ferritin (NF) and bovine serum albumin as evidenced ultrastructurally by the increase in accumulation of NF in the GBM. In addition, the permeability of the GBM to 125I-insulin, a macromolecule which is normally freely permeable, and the glomerular filtration rate (as determined by [3H]inulin clearance) were markedly reduced after the GBM had been clogged with NF in the presence of high molarity buffer, thereby indicating that clogging severely reduces the ability of the GMB to act as a selective filter. These findings are consistent with the idea that the sulfated GAGs of the GBM serve as anticlogging agents.

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Effects of weak bases on the degradation of endogenous and exogenous proteins by rat yolk sacs

Biochemical Journal ◽

10.1042/bj1880895 ◽

1980 ◽

Vol 188 (3) ◽

pp. 895-903 ◽

Cited By ~ 29

Author(s):

G Livesey ◽

K E Williams ◽

S E Knowles ◽

F J Ballard

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Normal Values ◽

Free Medium ◽

Weak Base ◽

Weak Bases ◽

Endogenous Proteins ◽

Hydrolysis Of

In rat yolk sacs incubated in vitro, the rates of degradation of endogenous [3H]leucine-labelled proteins and of pinocytically ingested 125I-labelled bovine serum albumin were both decreased in the presence of either ammonium, methylammonium or ethylammonium ions (0-20 mM) or much lower concentrations of chloroquine (0-500 microM). These effects were also accompanied by an inhibition of pinocytosis, as measured by the rate of uptake of 125I-labelled polyvinylpyrrolidone, and by a fall in the [ATP]/[ADP] ratio within the tissue. Re-incubation in inhibitor-free medium of yolk sacs previously exposed to a weak base restored pinocytic and proteolytic capacities, except for tissues exposed to chloroquine at concentrations above 0.1 mM (these appeared to be cytotoxic); an attendent rise in [ATP]/[ADP] ratios to near normal values was also observed. Weak bases, at concentrations that fully arrested the breakdown of 125I-labelled albumin, failed to inhibit by more than 45% the degradation of [3H]leucine-labelled endogenous proteins. Since 125I-labelled bovine serum albumin has been shown to be degraded entirely intralysosomally by yolk sacs, this suggests either that the hydrolysis of endogenous proteins is shared between lysosomes and some other site or that, unlike 125I-labelled albumin, some endogenous proteins can be degraded within lysosomes at abnormally high pH.

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THE NATURE OF ANTIGEN-ANTIBODY COMPLEXES FORMED IN RABBITS DURING AN IMMUNE RESPONSE TO BOVINE SERUM ALBUMIN

Journal of Experimental Medicine ◽

10.1084/jem.107.5.653 ◽

1958 ◽

Vol 107 (5) ◽

pp. 653-663 ◽

Cited By ~ 4

Author(s):

William O. Weigle

Keyword(s):

Immune Response ◽

Bovine Serum Albumin ◽

Serum Albumin ◽

Ammonium Sulfate ◽

Bovine Serum ◽

Ammonium Sulfate Precipitation ◽

Antibody Synthesis ◽

Circulating Antigen ◽

Antigen Antibody

The immune elimination of soluble BSA, following an intravenous injection, is accompanied by the appearance of circulating antigen-antibody complexes. The pattern of the appearance of circulating antigen-antibody complexes and the immune elimination of antigen probably depends on the amount of antigen injected, the rate of antibody synthesis, and perhaps, the quality of antibody produced. There is no relationship between the I* antigen-antibody complexes detected during the immune response in rabbits by ammonium sulfate precipitation and the material precipitated from immune sera as a result of treatment with alkali. Alkali-precipitable material present in the serum of rabbits at a time when I* antigen is also present contain at most only traces of the antigen.

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STUDIES ON THE IMMUNOLOGICAL SPECIFICITY OF AUTOLOGOUS GAMMA-GLOBULIN IN THE GLOMERULI OF RABBITS WITH EXPERIMENTALLY INDUCED GLOMERULONEPHRITIS

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o63-102 ◽

1963 ◽

Vol 41 (4) ◽

pp. 897-904

Author(s):

A. L. Sherwin ◽

A. Leznoff ◽

M. Richter ◽

B. Rose

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Specific Antibody ◽

Bovine Serum ◽

Gamma Globulin ◽

High Titer ◽

Fluorescent Antibody ◽

Fluorescent Antibody Technique ◽

Antibody Technique ◽

Immunological Specificity

Experimental glomerulonephritis was induced in rabbits by the intravenous administration of chicken antirabbit glomerulus serum (nephrotoxic serum). These rabbits were simultaneously immunized, either actively or passively, with bovine serum albumin or ovalbumin. Autologous gamma-globulin was identified in the glomeruli of these nephritic rabbits by means of the fluorescent antibody technique. It was not possible to demonstrate the presence of an anti-BSA or anti-OA component in the gamma-globulin even though these antibodies were present in high titer in the blood. This suggests that the autologous gamma-globulin present in the lesions is entirely specific antibody rather than an accumulation of serum gamma-globulin.

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ANTI-BOVINE SERUM ALBUMIN AND ANTI-ALPHA LACTALBUMIN IN THE SERUM OF CHILDREN AND ADULTS

PEDIATRICS ◽

10.1542/peds.35.4.571 ◽

1965 ◽

Vol 35 (4) ◽

pp. 571-588

Author(s):

Richard M. Rothberg ◽

Richard S. Farr

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Gamma Globulin ◽

Age Distribution ◽

Skin Tests ◽

Positive Skin ◽

Antigen Antibody ◽

Alpha Lactalbumin

1. Because precipitin, hemagglutination, and complement-fixation tests measure secondary manifestations of antigen-antibody interactions and are sometimes negative even after a primary antigen-antibody reaction has occurred in vitro, the incidence and amount of anti-bovine serum albumin (BSA) was evaluated in the sera from 900 children and adults by means of precipitating I131-labeled BSA-antibody complexes with 50% saturated ammonium sulfate. A similar study using I131-labeled alpha lactalbumin (ALA) was performed on 718 of this same group of sera. 2. Antibody to BSA was detected more frequently among children (75%) than among young adults 16 to 40 years of age (25%), or among older age groups (8%). 3. The incidence of detectable antibody to ALA had the same age distribution, but only half the frequency as anti-BSA. 4. In contrast to the near absence of antibody in the cord serum as measured by hemagglutination titers using red cells coated with milk proteins, most of the antibody detected in maternal sera in the present study was able to cross the placental barrier and was present in the cord sera. 5. The incidence of both anti-BSA or anti-ALA was the same in males and females. 6. If a given sera bound both IBSA and IALA, the anti-BSA activity was usually, but not always, greater than the anti-ALA activity. 7. No shared antigenicity was detected between IBSA and ovalbumin, insulin, protamine, diptheria, and tetanus toxoid, pertussis vaccine, poliomyelitis vaccine, and influenza vaccine. The apparent inhibiting effects of unlabeled bovine gamma-globulin and ALA on IBSA binding were probably due to trace amounts of BSA in these protein preparations. 8. BSA, ovalbumin, and bovine gamma-globulin had no detectable shared anti-genicity with IALA. 9. Positive skin tests to milk or BSA did not correlate with the anti-BSA levels measured in the serum. 10. The incidence of persons with anti-BSA and anti-ALA was comparable among the "patient" and "well" populations. Ten of the 31 sera with the greatest capacity to bind IBSA were from the "well" population, the remaining 21 sera were from children with a variety of disease states.

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HISTOLOGICAL AND SEROLOGICAL SEQUENCES IN EXPERIMENTAL HYPERSENSITIVITY

Journal of Experimental Medicine ◽

10.1084/jem.85.6.571 ◽

1947 ◽

Vol 85 (6) ◽

pp. 571-590 ◽

Cited By ~ 130

Author(s):

Clinton Van Zandt Hawn ◽

Charles A. Janeway

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Gamma Globulin ◽

Horse Serum ◽

Protein Solutions ◽

Pathological Lesions ◽

Intravenous Injections ◽

Foreign Proteins ◽

Antigen Antibody

1. Groups of normal rabbits were given, single intravenous injections of foreign proteins in doses of 1 gm. per kilo, bled at regular intervals for serologic studies, and sacrificed after varying lengths of time for pathological studies. The protein solutions used were of crystallized bovine serum albumin, bovine serum gamma globulin, and bovine serum. The experiments were planned, first, to correlate the sequence of pathological and immunological changes, and second, to compare the responses to two chemically and immunologically distinct plasma protein fractions and to the whole serum of the same species. 2. (a) The principal pathological lesions in rabbits given bovine serum were similar to those which have been previously observed following, the injection of horse serum and were characterized by widely dispersed but segmental acute inflammatory lesions of the arteries. These lesions were at their height 2 weeks after injection and showed marked repair at 4 weeks. (b) Crystallized bovine serum albumin produced lesions almost exclusively confined to the arteries which were at their height at 2 weeks, were healing at 3, and healed by 4 weeks. The lesions were less numerous and less intense than in animals given whole serum and were only found in some of the animals. (c) Bovine serum gamma globulin elicited quite different histologic sequences. The most striking lesions involved the glomeruli of the kidneys, and to a lesser degree, the heart. Lesions in the liver and joints were present but less conspicuous, and arterial lesions were rare and slight in degree. The lesions not only differed from those in rabbits given albumin in distribution but in timing, since they were most widespread and acute at 1 week and were healing at 2 weeks after injection. Moreover, lesions were observed in almost every animal. 3. Results of immunological studies were consistent with the interpretation that the pathological lesions were due to an antigen-antibody reaction in the tissues, as shown by the following: (a) Acute lesions were only observed when antigen was present and before antibody appeared in the circulation. (b) Healing of lesions was only observed (with one exception) when antigen had almost or completely disappeared from the circulation, usually with the appearance of antibody. (c) There was a correlation between the rapidity of evolution of the lesions and the rapidity with which the antigen disappeared from the circulation. (d) There was a rough correlation between the proportion of animals showing lesions and the proportion developing antibodies after the injection of a particular protein solution.

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