scholarly journals ELECTRON MICROSCOPIC LOCALIZATION OF THE NEPHROTOXIC ANTIBODY IN THE GLOMERULI OF THE RAT AFTER INTRAVENOUS APPLICATION OF PURIFIED NEPHRITOGENIC ANTIBODY-FERRITIN CONJUGATES

1968 ◽  
Vol 127 (5) ◽  
pp. 867-878 ◽  
Author(s):  
Arnold Vogt ◽  
Hermann Bockhorn ◽  
Keniti Kozima ◽  
Masamichi Sasaki

Nephritis in rats was induced by intravenous injection of purified ferritin-conjugated rabbit and duck nephrotoxic globulin. Using the fluorescent antibody technique, the same capillary pattern was found as that in glomeruli of rats receiving uncoupled nephrotoxic globulin. Electron microscopy revealed a heavy accumulation of the basement membrane-fixed antibody almost exclusively at the endothelial side. A higher concentration of ferritin was demonstrable in the peripheral basement membrane. The once-fixed antibody remained at the site of reaction though decreasing with time. The half-disappearance time seemed to be shorter than that of the uncoupled nephrotoxic globulin. No difference in localization was observed between rabbit and duck antibody. At least 40 basement membrane-fixed antibody molecules from the rabbit per 3000 mµ2 of filtration surface were needed to cause immediate nephritis. To induce nephritis using duck antibody, a greater number of basement membrane-fixed antibody seemed to be necessary. No evidence of specific reaction with constituents of glomerular cells was obtained.

1962 ◽  
Vol 115 (5) ◽  
pp. 929-936 ◽  
Author(s):  
Giuseppe A. Andres ◽  
Councilman Morgan ◽  
Konrad C. Hsu ◽  
Richard A. Rifkind ◽  
Beatrice C. Seegal

Ferritin-conjugated antibody has been used to identify by electron microscopy the sites at which nephrotoxic globulins localize in rat kidney during acute experimental glomerulonephritis. Antibody was concentrated in the glomerular basement membrane and in basement membrane-like material contained in distended cisternae of the endoplasmic reticulum. These data confirm and amplify, at the ultrastructural level, the results of studies obtained with the fluorescent antibody technique, and are consistent with the hypothesis that the cisternae and capillary basement membrane possess common proteins.


1967 ◽  
Vol 33 (2) ◽  
pp. 307-318 ◽  
Author(s):  
Paul K. Nakane ◽  
G. Barry Pierce

Enzymes, either acid phosphatase or horseradish peroxidase, were conjugated to antibodies with bifunctional reagents. The conjugates, enzymatically and immunologically active, were employed in the immunohistochemical localization of tissue antigens utilizing the reaction product of the enzymatic reaction as the marker. Tissues reacted with acid phosphatase-labeled antibodies directed against basement membrane were stained for the enzyme with Gomori's method, and those reacted with peroxidase-labeled antibody were stained with Karnovsky's method. The reaction products of the enzymes localized in the basement membrane. Unlike the preparations of the fluorescent antibody technique, enzyme-labeled antibody preparations were permanent, could be observed with an ordinary microscope, and could be examined with the electron microscope. In the latter, specific localization of antibody occurred in the basement membrane and in the endoplasmic reticulum of cells known to synthesize basement membrane antigens. The method is sensitive because of the amplifying effect of the enzymatic activity. The ultrastructural preservation and localization were better with acid phosphatase-labeled antibody than with peroxidase-labeled antibody, but acid phosphatase conjugated antibody was unstable and difficult to prepare. Peroxidase-antibody conjugates were stable and could be stored for several months at 4°C, or indefiniely in a frozen state.


1981 ◽  
Vol 18 (5) ◽  
pp. 672-683 ◽  
Author(s):  
W. C. Hall ◽  
J. D. White ◽  
R. A. Kishimoto ◽  
R. E. Whitmire

Athymic nude mice and euthymic littermate controls were exposed to 104 Coxiella burnetii organisms by small-particle aerosol. Antibody response with and without 2-mercaptoethanol treatment of serum was determined at various intervals after infection and serial kills were done to determine morphologic changes in both mouse phenotypes. Total antibody titers determined by the indirect fluorescent antibody technique to phase I and phase II C. burnetii were identical for both groups of mice. Microagglutinin titers determined on days 28 and 33 were abolished by 2-mercaptoethanol treatment of serum from both phenotypes, indicating that the antibody probably resided in the IgM fraction. Microscopically, the reaction to C. burnetii infection was similar in nude and euthymic mice on days 7 and 14. Later, the number and size of lesions attributable to Q fever diminished in euthymic mice. Infection was progressive in nude mice, with macrophage infiltration of most tissues, especially spleen and liver. Numerous rickettsiae were seen by immunofluorescence and electron microscopy in phagocytic vesicles of macrophages, many of which were dilated, giving the macrophage a vacuolated appearance. Results suggest that clearance of C. burnetii infection in mice is dependent upon thymus-derived lymphocytes.


1952 ◽  
Vol 95 (1) ◽  
pp. 105-118 ◽  
Author(s):  
Willard C. Schmidt

A method was developed for the extraction of the group A streptococcus polysaccharide employing pepsin digestion of ground streptococcal cells. This method did not result in the isolation of polysaccharide with chemical and physical chemical properties different from those exhibited by preparations extracted with hot formamide. Studies of the chemical composition of this polysaccharide demonstrated it to be composed chiefly of rhamnose and glucosamine monosaccharide units in the approximate ratio of five moles of rhamnose to two moles of glucosamine. The fate of the polysaccharide after intravenous injection into mice was studied using the fluorescent antibody technique. It was found to be rapidly eliminated by the kidney. The presence of the polysaccharide in the renal tubular epithelial cells during the excretory phase was the only evidence of its cellular localization that could be detected under the conditions of these experiments.


Author(s):  
Veronika Burmeister ◽  
R. Swaminathan

Porphyria cutanea tarda (PCT) is a disorder of porphyrin metabolism which occurs most often during middle age. The disease is characterized by excessive production of uroporphyrin which causes photosensitivity and skin eruptions on hands and arms, due to minor trauma and exposure to sunlight. The pathology of the blister is well known, being subepidermal with epidermodermal separation, it is not always absolutely clear, whether the basal lamina is attached to the epidermis or the dermis. The purpose of our investigation was to study the attachment of the basement membrane in the blister by comparing scanning with transmission electron microscopy.


2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.


Science ◽  
1964 ◽  
Vol 145 (3635) ◽  
pp. 943-945 ◽  
Author(s):  
G. C. Brown ◽  
H. F. Maassab ◽  
J. A. Veronelli ◽  
T. J. Francis

1965 ◽  
Vol 13 (3) ◽  
pp. 155-160 ◽  
Author(s):  
H. J. KEUTEL

Fluorescent labeled antibodies were used for the demonstration of uromucoid. This urine specific mucoprotein is demonstrably present only in the epithelial cells of the proximal segments of the normal human renal tubules and in the matrix of human kidney stones of all the common crystalline compositions.


Sign in / Sign up

Export Citation Format

Share Document