Anti-kidney Serum Research Articles

Glomerular basement membrane in experimental nephrosis: x-ray diffraction and electrophoretic studies.

Basement membrane obtained from glomeruli of normal and nephrotic (anti-kidney serum) rats has been examined by microscope electrophoresis and X-ray diffraction techniques. Normal membrane had a mobility at pH 7.35 of 0.93 ± 0.12 <i>µ</i>/s/V/cm; the X-ray diffraction pattern resembled that of collagen, with an additional ring corresponding to a spacing at 2.25 Å. The membrane from nephrotic rats had a mobility of 0.69 ± 0.027 <i>µ</i>/s/V/cm, significantly lower than normal; the mobility was also decreased at pH of 2.25 and 8.75. The X-ray diffraction pattern showed several reflections corresponding to short spacings, not seen in the normal pattern, some of which disappeared on treatment of the membrane with collagenase. It is suggested that the changes in mobility and X-ray diffraction may be the result of loss of phospholipid and that the changes in diffraction pattern result from an increased ordering within the crystal lattice of both collagen and non-collagenous components.

Glomerular Basement Membrane in Experimental Nephrosis: Chemical Composition

Basement membrane prepared by ultrasonic vibration of isolated glomeruli was obtained from normal rats and from rats with anti-kidney serum nephrosis. The membrane was found to contain collagen, lipid

A comparative study of kidney and muscle membrane antigens in the human and rat

Argyrophile, PAS-positive, membranous and fibrillar structures of the diverse organs share a number of common antigens, referred to as “membrane antigens.” The nephrotoxic serum antigens belong to this group. The present communication reports on a comparative study of muscular and renal membrane antigens in the human and rat Antisera obtained by heteroimmunization with organ homogenates contain a variety of antibodies; the homologous antigens are partly common to many organs and partly organ-specific. The number of membrane antigens is unknown, since they cannot be isolated in pure form. Striated muscle contains fewer membrane antigens than the kidney, but at least one nephrotoxic serum antigen is present. The injection of rabbit anti-kidney and anti-skeletal muscle serum in the rat causes glomerulonephritis. Whereas glomerular localization of rabbit antibody globulin is readily demonstrable in both cases; a host response, i.e., glomerular fixation of the rat's own gammaglobulin, is found only in animals given anti-kidney serum. Furthermore, the nephrotoxicity of anti-kidney serum cannot be neutralized by absorption with muscle material. It is postulated that renal and muscular nephrotoxic serum antigens differ in some as yet unknown qualitative or quantitative properties.

Electron Microscopic Studies on Antigen-Antibody Reaction in the Kidney and its Similar Reaction : 1) Glomerular Changes Caused by Anti-Kidney Serum in Pregnant Rabbits

Electron Microscopic Studies on Antigen-Antibody Reaction in the Kidney and its Similar Reaction : 1) Glomerular Changes Caused by Anti-Kidney Serum in Pregnant Rabbits

ADIPOSE TISSUE IN EXPERIMENTAL NEPHROTIC SYNDROME.

Epididymal adipose tissue of aminonucleoside-treated rats, investigated 3 to 6 days after induction of the nephrotic syndrome, had low glycogen levels and showed impaired esterification of free fatty acids and assimilation of lipoprotein triglyceride and markedly reduced liberation of lipoprotein lipase. These results were found to be influenced by the inadequate food intake of the acutely nephrotic animals and comparable to the values of control rats fasted for 2 days. On return to adequate nutrition, which occurred 12–20 days after aminonucleoside treatment, adipose tissue glycogen and free fatty acid assimilation returned toward normal levels but lipoprotein-lipase liberation remained below normal. In rats rendered nephrotic by antikidney serum, the assimilation of free fatty acids and lipoprotein-triglyceride by adipose tissue was impaired in spite of only minor reduction in food consumption. The results indicate that the defective metabolism of adipose tissue in nephrotic animals may be contributory to the nephrotic hypertriglyceridemia.

Lung localizing antibodies in anti-lung and anti-kidney serum.

SummaryLocalization of antibodies in lungs was examined by fluorescent antibody technic after injection of rabbit anti-rat lung and anti-rat kidney serum into rats. Although both sera gave positive staining in lungs, anti-lung serum contained significantly higher amount of lung localizing antibodies than anti-kidney serum. The localizing components in both sera seemed to be fixed in the same region of alveolar walls.

THE PATHOLOGIC EFFECTS OF INTRAVENOUSLY ADMINISTERED SOLUBLE ANTIGEN-ANTIBODY COMPLEXES

The intravenous administration to rats of soluble antigen-antibody complexes in antigen excess resulted in acute glomerulonephritis. This occurred with both rabbit antiovalbumin and rabbit anti-BSA systems. The rats so treated regularly showed proteinuria and elevation of blood urea nitrogen. These findings are compared with those reported in rats injected with anti-kidney serum.

Lipoprotein lipase metabolism in experimentally nephrotic rats.

SummaryHeparin-activated lipolytic mechanisms were studied in vitro in rats with nephrosis experimentally induced by injection of antikidney serum. The nephrotic rat was found normally capable of releasing lipoprotein lipase following heparin administration. The nephrotic rat's plasma also was found normally capable of supporting lipoprotein lipase-induced hydrolysis of a triglyceride substrate.

Renal glomerular lesions induced by rabbit antirat collagen serum in rats prepared with adjuvant.

Renal glomerular lesions were induced by rabbit serum containing antibody to rat collagen injected intravenously into rats prepared with subcutaneously administered Freund adjuvant. Neither the anti-collagen serum nor the adjuvant alone induced the lesion. The lesions were characterized by diffuse glomerular injury with swelling, shredding, and fusion of the basement membranes, crescent formation, cellular proliferation, numerous multinuclear giant cells, and capillary hyaline thrombi. Various rabbit antisera, including those against fish collagen or rat serum failed to induce the renal lesion when substituted for anti-rat collagen serum. Also, anti-rat collagen serum absorbed with its homologous antigen, native rat collagen, failed to induce the lesion. Although complete adjuvant, i.e. with mycobacteria, in which normal serum was incorporated enhanced the glomerular lesion which resulted from intravenous injection of anti-collagen serum, the incomplete adjuvant without serum was sufficient. Comparison of the renal lesions induced by anti-collagen serum with nephrotoxic nephritis induced in rats by rabbit anti-kidney serum showed that they differ histologically. Also the antisera used to produce these two renal lesions differ immunologically. Antibodies to normal rabbit serum developed in rats injected intravenously with rabbit anti-rat collagen serum after preparation with adjuvant, but not when adjuvant was omitted. The pathogenesis of the renal injury is discussed as a manifestation of an antigen-antibody reaction, with nephritis occurring only after the adjuvant-stimulated antibody to the rabbit globulin has been formed in the rat and has reacted with the rabbit anti-rat collagen already fixed by its homologous antigen in the kidney.

Adenosinetriphosphatase Activity in the Cell Membranes of Kidney Tubule Cells

This cytochemical study demonstrates high levels of apparent ATPase activity in the infolded cell membranes of the proximal tubules (dog, rat, human, mouse, monkey, and opossum) and ascending loops of Henle (dog, rat, human and, to a variable degree, mouse). Electron microscopy has shown (see Rhodin (1)) that these membranes separate adjacent cells where they interlock with one another by multiple cytoplasmic lamellae containing oriented mitochondria. The significance of the high ATPase activity is considered in relation to possible movements of the membranes and to "active transport" believed to occur there. In the rat, enzyme activity in the proximal tubule membranes does not survive formol-calcium fixation, and it is therefore necessary to use unfixed sections for its demonstration. However, in edematous rats with experimental nephrosis (induced by the injection of aminonucleoside or with antikidney serum) marked ATPase activity is exhibited in these membranes even after formol-calcium fixation. When proximal tubule or Henle loop cells of the dog acquire an altered metabolism, as indicated by accumulated lipide spheres or by "droplets," the infolded ATPase-rich membranes are no longer demonstrable.

Relationship between concentrations of albumin and lipids in plasma of experimentally nephrotic rats.

Correlative studies were made of the concentrations of albumin and lipids in the plasma of rats made nephrotic by injection of antikidney serum. It was found that the increased concentrations of plasma triglycerides, cholesterol and phospholipid in such rats were inversely correlated with the magnitude of their lowered plasma albumin content, rather than with the amount of antikidney serum that was injected. The results are consistent with the belief that a causal relationship exists between the deficiency of albumin and the increased lipid content of nephrotic plasma.

Localizing Properties of Anti-Placenta Serum

Abstract Antisera prepared against placenta are known to be cytotoxic (1–5). Dobrowolski (1) in 1903 showed that specific anti-placenta sera interrupted pregnancy in guinea pigs and rabbits. Seegal et al. (2–3) found that antisera prepared in rabbits against rat placenta when injected into rats cause interruption of pregnancy and also nephritis. Rabbit anti-dog-placenta serum produced nephritis in dogs but not abortion (4–5). Assuming that cytotoxic antibodies must localize in the tissue affected, one would expect accumulation of anti-placenta antibody in the kidney and placenta of rats injected with antiplacenta antiserum. Therefore, we made a study in pregnant and nonpregnant rats of the localizing properties of a known cytotoxic rabbit anti-rat-placenta sera and the results are reported here. We also compared the localization of antibodies from anti-rat-kidney serum with those of anti-placenta serum. It was found that the anti-placenta serum and the anti-kidney serum each contained antibodies capable of localizing in both the kidneys and the placenta.

The production of glomerulonephritis by immunologic methods

1. 1. Specific antikidney, antiglomerular, or antiplacenta serum, when injected in the rat or dog, produces acute glomerulonephritis, which may be fatal within a few days, may heal, or may progress to chronic nephritis resulting in death from renal failure months or years later. Duck antirabbit-kidney serum injected in rabbits produces a similar result. 2. 2. The clinical course and pathologic lesions of this experimental disease resemble those of human nephritis. 3. 3. In the rat, high-titered specific antisera may produce a nephrotic syndrome characterized by edema, hypercholesterolemia, hypoproteinemia, and massive proteinuria. 4. 4. Between the clinical picture of acute nephritis and the onset of chronic nephritis in the rat, there may be an interval of several months during which time the animal appears normal. 5. 5. Nephritis produced in rats, rabbits, or dogs by the injection of specific duck or chicken antikidney serum often has a latent period of several days. 6. 6. Evidence has been presented that, at least in the rabbit, the onset of nephritis after a latent period is the result of a reaction between the injected antikidney foreign protein (fowl gamma globulin) attached to kidney parenchyma and the antibodies produced by the rabbit to this antigen. 7. 7. The lesions of delayed nephritis in the rabbit, rat, or dog are similar to those which occur in nephritis with no latent period. 8. 8. Evidence has been developed from chemical and histochemical studies of the glomerulus of the dog and rat which indicates that the basement membrane of the glomerulus may be the source of the antigen inciting production of the nephrotoxic antibody. 9. 9. The new immunologic and histochemical techniques which have been used in the study of experimental nephrotoxic nephritis may in the future give data on the mechanism of human nephritis.

Histologic changes in jaws and teeth of rats following nephritis, adrenalectomy, and cortisone treatment

We have described the effects of certain systemic alterations on the jaws and teeth of the rat. A loss of supporting bone was observed in the jaws of rats subjected to cortisone alone or adrenalectomy alone. A similar loss of supporting bone was observed in the jaws of rats subjected to a combination of factors—adrenalectomy, antikidney serum, and cortisone. There was also a marked change in the bone pattern of these rats. Certain vascular changes in the pulps of the molar teeth of these rats were described. After adrenalectomy, the pulpal blood vessels were dilated and there was hemorrhage. Rats given cortisone showed dilated blood vessels in the molar pulps. Similar changes were observed in rats which were adrenalectomized and then given cortisone plus antikidney serum. Rats given antikidney serum alone did not show a severe dilation of the pulpal blood vessels. The factors involved in these changes were discussed.

Lipemia clearing by hyaluronidase, hyaluronate, and desoxycorticosterone, and its inhibition by cortisone, stress, and nephrosis.

Summary1. Partially depolymerized hyaluronic acid released a lipemia clearing factor following intravenous, subcutaneous and oral administration to rats. Hyaluronidase and DC A which were administered only to rats hypodermically were also effective. 2. Plasma from rats treated with cortisone prevented the lipemia clearing by heparin, DCA and partially depolymerized hyaluronic acid. The effect of exposure to cold was tested only against heparin lipemia clearing and was also found to inhibit it. 3. Release of a lipemia clearing inhibitor appears to be another manifestation of the general adaptation syndrome. 4. Plasma from rats made nephrotic by administration of low doses of anti-kidney serum also had an inhibitor of the clearing factor, and those given larger doses of anti-kidney serum had in addition gross lipemia suggesting that pathological lipemia may be preceded by the appearance of the inhibitor. 5. The lipemia clearing factor was inactivated by incubation with hyaluronidase in vitro suggesting that...

Arresting effect of heparin upon experimental nephrosis in rats.

SummaryHeparin administration decreases the hyperlipemia and hypercholesteremia of rats with established nephrosis, induced by injection of rabbit antirat kidney serum. Heparinization during the induction phase of the nephrotic state appears to suppress or prevent ascites and edema, and to reduce markedly the degree of lipemia, hypercholesteremia and lipemic-turbidity of the plasma. Possible mechanisms of this arresting effect of heparin are discussed briefly and it is suggested that heparin suppresses the causal mechanism whereby nephrosis is induced in rats by injection of anti-kidney serum.

The In-Vitro Purification of Tissue Localizing Antibodies

Summary The purification of antibodies in vitro was carried out by adsorbing radioiodinated anti-liver, anti-kidney, anti-lung and anti-spleen sera with a blood vessel fraction or cellular fraction of an organ and then eluting the adsorbed radio-activity by heat at 60°C. It was found that the eluates obtained from blood vessel fractions of liver, kidney and lung had different localizing properties from the eluates of the cellular fractions of the same organs. It was also found that eluates could be prepared which would localize preferentially in either liver, kidney, lung or spleen by purifying the proper anti-tissue serum with a homologous tissue fraction. Antibodies from anti-lung, anti-liver and anti-spleen sera localized to the greatest concentration in the homologous organ when the purification was carried out with the cellular fractions of lung, liver and spleen respectively. With the anti-kidney serum the best results were obtained with the kidney blood vessel fraction.

The Zone of Localization of Antibodies IX.

Summary Rats injected with the radioiodinated globulin fraction of anti-lung serum exhibited specific localization of radioactivity in their lungs, and also in their kidneys. Although the localization in kidney of anti-lung serum was quite similar in extent to the localization of anti-kidney serum, the localization in lung was quite different for the two antisera. Absorption by acetone extracted kidney sediment removed more kidney-localizing than lung-localizing activity, suggesting that there are substances in lung which are different serologically from any components in kidney. The kidney-localizing component of anti-lung serum was found to be similar to that of anti-kidney serum: both localized clearly in glomeruli (radioautography), decayed in kidney tissue with a “half-life” of about 25 days, and were absorbed in vitro by a fraction of kidney homogenate which was insoluble in saline, alcohol, acetone and ether. It is concluded that there are substances in lung and kidney which are similar, or perhaps identical, and which are the antigens responsible for the development of kidney-localizing antibody. The findings are discussed in relation to glomerulonephritis.

The Zone of Localization of Antibodies. IV

Summary Antisera were prepared in rabbits against kidney tissue and plasma from the Akm strain of mice and against ovalbumin. The globulin fractions of these sera were radioiodinated and injected into mice of several strains and into rats. It was found that radioactivity localized in the kidneys of the animals receiving the antikidney serum, but not in the kidneys of those receiving the antiplasma serum or antiovalbumin serum. Radioiodide also showed no such localization. This demonstrated that there was some material, presumably antibody molecules, in the antikidney serum and not in the control sera, that localized in the kidney. The localization of antikidney serum was not strain specific since it took place in mice of other strains. It also took place on the injection of rats with the anti-mouse-kidney preparation. The duration of the radioactivity in the kidneys of the mice receiving the antikidney serum was measured over a period of 54 days and the radioactivity carrier, presumably the radioiodinated antibody, had a half life of 20 days. Evidence was also found for the presence of a factor in the antikidney serum, presumably antibody to common antigens, which cross reacted with and localized in liver and spleen, but to a much lower extent than in kidney. In liver there was an initially rapid decrease in radioactivity followed by a subsequent slow decrease with a half life of 20 days. The long half life of the antibody in the kidney and liver was probably due to the inability of this tissue to metabolize the antibody. The half lives of the radioactive carriers in other tissues were blood, 2.3 days; spleen, 2.6 days; and thyroid 6.8 days. Acknowledgement. This work was carried out with the assistance of Miss Beila Sherman, Miss Rhoda Michaels, Mr. Arthur Silverstein and Mr. Richard Clark.

Desoxycorticosterone acetate studies on the reversibility of its effect on blood pressure and renal damage in rats.

In previous studies it has been reported that the administration of desoxycorticosterone acetate (DCA) to rats, when accompanied by a liberal intake of sodium chloride, leads to cardiac and renal enlargement (Selye, 1940; Green, Coleman and McCabe, 1948; Knowlton, Stoerk, Seegal and Loeb, 1946), renal damage (Selye 1943) and to the development of hypertension (Grollman, Harrison and Williams 1940; Selye and Hall, 1943). The alterations produced by this steroid are much intensified in the presence of pre-existing renal damage. With the experimental conditions employed in this laboratory, hypertension following DCA injections has been observed only in rats previously rendered nephritic by a cytotoxic serum (a rabbit anti-rat kidney serum). In these hypertensive animals, a more rapid progression in the course of the pre-existing nephritic process has been found. The present study was undertaken in an attempt to determine whether the hypertension in rats given anti-kidney serum and DCA was a phenomenon depend...