[Immunofluorescence of the kidney].
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Biomedical subjects
Publications and source records attributed to F Shimizu.
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A water-soluble nitrosourea, 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1-(2-chloroethyl)-1-nitrosourea hydrochloride (ACNU, NSC-245382), was tested for its antitumor activity against some kinds of transplantable mouse tumors. The compound was markedly active against myeloid leukemia C1498, plasmacytoma X5563, Ehrlich ascites carcinoma, and mammary tumor FM3A43, and moderately active against mammary tumor MM102 and meningeal sarcoma MS147. It appears that ACNU has a broad antitumor spectrum.
Antibodies to glomerular basement membrane (GBM) of the rat could be fractionated according to their avidity by elution from trypsin-digested GBM (bound to Sepharose) with increasing concentrations of KSCN. The percentage of kidney-fixing antibody in each fraction and the degree of proteinuria induced as determined 24 h after injection increased with the avidity of the antibody fraction when equal doses were administered.
Mouse nonimmune peripheral blood lymphocytes (PBL) plus antibody to herpes simplex virus inhibited virus dissemination in herpes simplex virus-infected 3T3 cell culture as revealed by development of viral plaque, size of immunofluorescent foci, appearance of polycaryocytes, and viral infectivity appearing in the culture. These nonimmune PBL did not act alone in inibiting virus dissemination, but did act synergistically in combination with antibody. The ratio of PBL to target monolayer cells needed to produce this effect was 20. Splenic lymphocytes had weak activity, whereas thymocytes were without effect, even in the presence of antibody. Neither interferon nor lymphotoxin was detected in this lymphocyte-mediated response. These findings support previous observations, based on cytotoxicity assays, that antibody-dependent cellular immune processes could be important in control of and recovery from herpesvirus infection.
Immunofluorescence studies revealed the age-associated presence of mouse IgG, with or without beta1 C in renal glomeruli of various strains of laboratory mice. These were classified into two types: (1) IgG demonstrated without beta1 C and the fluorescence of which was easily decreased by prewashing with PBS. (2) IgG demonstrated almost always with beta1 C and the fluorescence of which remained unchanged by prewashing with PBS. The deposition of immunoglobulin seems to be closely connected with the immune state of animals including the phagocytic function of the mesangium.
The infectivities of herpes simplex virus types 1 and 2 were inactivated by silver nitrate at concentrations of 30 muM or less, which did not affect at all the infectivities of hemagglutinating virus of Japan, vesicular stomatitis virus, poliovirus, vaccinia virus, and adenovirus. The inactivated virus retained the capability of adsorbing to the cell, with an adsorption kinetics quite similar to that of intact virus, and of inducing the concanavalin A agglutinability in the infected cells, whereas it lost completely the capability of producing viral antigens and other cytopathic changes.
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Surface markers and organ distributions of B lymphocytes bearing surface immunoglobulins (SIg B) and lymphocytes lacking SIg but carrying Fc receptors (SIg-Fc+), and the variations of both the cell populations in the autoimmune diseases were studied. Peripheral blood lymphocytes contained approximately 9% SIg B and 22% SIg-Fc+ cells. Both the populations of SIg B and SIg-Fc+ cells were included in the cells that formed no E-rosettes. When previously treated with neuraminidase, however, the SIg-Fc+ cells formed E-rosettes, whereas SIg B showed no ability to form rosettes. C receptors as detected by human complement were found in the large portions of SIg B but in a minority of the SIg-Fc+ cell population. The tissues that contained the highest concentrations of SIg B and SIg-Fc+ cells were the spleen and peripheral blood, respectively. In the patients with an autoimmune disease in active stages SIg B cells in the peripheral blood markedly increased and, reversed, SIg-Fc+ cells decreased; in contrast, in remission of the diseases SIg B cells dropped down and SIg-Fc+ increased up to their normal levels. A possible relationship between the SIg B and SIg-Fc+ cells in regard to the differentiation of B lymphocytes was presemed.
Some basic problems in immunofluorescence were reexamined, limiting the object to kidneys. As to thickness of sections, 4 mum was demonstrated to be the most reasonable also to kidney specimens. Treatment of PBS for a short time before staining was recommended not only for cleanup effect but also for the possibility to disclose covered antigenic determinants. It was also established that fluorescent antibody with adequate F/P molar ratio, as well as with strict specificity, should be used, in order to prevent "false positives".
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Immunoferritin conjugates consist of conjugates of different size. They can be separated in 4.5% polyacrylamide gel electrophoresis into at least 4 bands. The amount of the smallest 1:1 conjugate, in which one antibody molecule is linked to one ferritin molecule, is highest in coupling products with low overall yields of conjugated ferritin. Therefore relatively mild reaction conditions are recommended. With increasing size the antibody binding capacity of the conjugate is reduced.
A water-soluble nitrosourea, 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1-(2-chloroethyl)-1-nitrosourea hydrochloride (ACNU), was tested for antitumor activity against lymphoid leukemia L-1210 in BDF1 mice. The results obtained were as follows: (1) ip--ip system: By daily administrations optimal dose for 12 days, MED (ILS40), and therapeutic ratio were found to be 18 mg/kg, 1.3 mg/kg, and 14, respectively, and by a single injection, those were found to be 46 mg/kg, 4.2 mg/kg, and 11, respectively. The therapeutic ratio of ACNU was the highest of all the drugs tested. At the neighboring dose level to optimal dose most of leukemic mice survived more than 60 days. (2) ip--iv system: Remarkable prolongation of life-span was observed by a single injection. Majority of mice administered more than 32 mg/kg of ACNU survived more than 60 days. (3) ip--po system: Remarkable prolongation of life-span was shown by a single administration and most of leukemic mice given more than 46 mg/kg of ACNU survived over 60 days. (4) iv--ip system: By a single administration of ACNU remarkable prolongation of life-span was seen and majority of leukemic mice were rescused at doses of 32 mg/kg or more. (5) iv--iv system: Result almost similar to iv--ip system was obtained. (6) iv--po system: The result in this system was similar to that in ip--po system. Thus, the compound was highly effective in leukemic mice by either its parenteral or oral administration.
With four clinical cases of contracted kidney in cats, granular to linear depositions of IgG and beta1C were demonstrated mainly in the capillary walls and mesangium of the glomeruli by means of immunofluorescence technique. The tubular basement membrane was also positive for cat IgG. These findings indicated that some immunological mechanisms might have been involved in the feline advanced renal lesions, as suggested in human cases. However, attempts were failed to detect some pathogenic agents for cats by immunofluorescence studies.
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A method, by which the more precise localization of immunofluorescent deposits in the glomerulus is easily demonstrated with the fluorescence microscopy, was described. By means of this method, the deposits even in the Bowman's space could be identified, as well as the deposits in the subepithelial, subendothlial and mesangial regions. From this result, it was suggested that this method should be widely used for the immunofluorescence studies on human renal biopsied sections, in order to elucidate the relationship between the localization of deposits and renal diseases.
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