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Results for “Immunoenzyme Techniques”

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At least 19 recordsLinked to original sources
PubMed1978

Immunohistochemical studies on human gastric mucosa. Procedures for routine demonstration of gastric proteins by immunoenzyme techniques.

Two different fixatives were applied to human gastric mucosa for the study of antigenic marker substances. The first consists of 96% ethanol and 1% acetic acid (EA method), the second of 4% formaldehyde, 0.5% picric acid and 0.25% glutaraldehyde (FPG method). Samples of resected gastric specimens were fixed, dehydrated and cleared in benzene and embedded in paraplast. The morphology of gastric tissue was well preserved by both methods and permitted the simultaneous application of classical staining procedures and the immunoenzyme peroxidase technique for the demonstration of antigenic substances. The following marker substances could be demonstrated: Pepsinogen I and II group, surface epithelial antigen, parietal cell antigen, chief cell antigen, antral mucous cell antigen, carcinoembryonic antigen, goblet cell antigen and common site antigen of leucocytes. Various factors responsible for nonspecific reactions, such as endogeneous peroxidase activity and protein interactions were studied. The latter were circumvented by the use of highly purified antibodies or immunoglobulin fractions. The EA method proved to be the method of choice for future routine application of combined classical histology and immunoenzyme histology in gastric and intestinal diseases.

Antigens↗
PubMed1977

Staining of human pituitary glands with lead hematoxylin in comparison with other histochemical procedures, including the immunoenzyme technique.

Human pituitary glands and pituitary adenomas, obtained by surgery or at autopsy, have been stained with lead hematoxylin and the results were compared with those of other histochemical procedures, including the immunoperoxidase technique. Lead hematoxylin positivity was found in the cytoplasm of melanocorticotroph, thyrotroph as well as gonadotroph cells, indicating that lead hematoxylin has no selective affinity for melanocorticotroph cells. Lead hematoxylin positive cells also stained with the PAS technique. Lead hematoxylin positivity was not revealed in growth hormone cells or in prolactin cells. Basophilic adenomas, composed of melanocorticotroph cells and associated with Cushing's disease or with Nelson's syndrome or unaccompanied by hormonal hypersecretion, showed intense lead hematoxylin positivity. Thus, no correlation was evident between lead hematoxylin positivity and secretory activity. Acidophilic or chromophobic adenomas consisting of growth hormone cells, prolactin cells, stem cells, undifferentiated cells or oncocytes, however, were negative. It can be concluded that the lead hematoxylin staining techniques a reliable, easy and reproducible histochemical method, and its use can be recommended in studies related to adenohypophysial cytology.

Adenoma↗
PubMed1977

An immunoenzyme histochemical technique for the detection of platelet antibodies from the serum of patients with idiopathic (autoimmune) thrombocytopenic purpura (ITP).

The resent report describes a simple, reproducible, semi-quantitative immunohistochemical assay for the detection of antiplatelet antibody. Evaluation of the technique in 10 children with active idiopathic (autoimmune) thrombocytopenic purpura (ITP), seven children with thrombocytopenia due to other causes, and 12 normal individuals revealed that the assay consistently and reliably distinguished children with ITP from the other groups. Individuals who had had multiple platelet transfusions and individuals with systemic lupus erythematosus (SLE) also had antiplatelet antibodies detectable using this technique although the levels were less than those of individuals with ITP. The method can be used effectively to monitor the course of ITP and the effects of therapy.

Adolescent↗
PubMed1979

Bunyavirus development in arctic and Aedes aegypti mosquitoes as revealed by glucose oxidase staining and immunofluorescence.

Northway virus replication has been detected in salivary glands of wild-caught Culiseta inornata and Aedes communis mosquitoes from the western Canadian Arctic after incubation at 4 degrees C for 9 to 11 months, and after incubation at 13 degrees C for 3 to 4 months after they received virus by oral ingestion or intrathoracic injection. Aedes hexodontus supported Northway virus replication after incubation at 13 degrees C for one month after intrathoracic injection. Aedes aegypti supported Northway virus replication after incubation at 13 degrees C or 23 degrees C for 6 to 28 days following intrathoracic injection. A larval isolate of California encephalitis virus (snowshoe hare subtype) multiplied in all 3 species of arctic mosquito after incubation at 13 degrees C for 1 to 3 months after virus was administered by oral ingestion or intrathoracic injection. Virus was detected in salivary glands of Cs. inornata after 329 days incubation at 4 degrees C after intrathoracic injection. Bunyavirus antigens in salivary glands of arctic and domestic mosquitoes were detected by the glucose oxidase immunoenzyme technique somewhat less frequently than by assay for virus infectivity.

Aedes↗
PubMed1978

Liver-specific and shared cell membrane antigens. Studies by light- and electron microscopy.

Liver-specific and shared saline-insoluble cell surface antigens were localized by immunofluorescence as well as by light- and electron microscopic immunoenzyme techniques. Antisera against purified mouse liver cell membranes were surface membrane but not organ-specific. Variable quantities of shared antigens were present in endoderm- and mesoderm-derived organs but not in ectodermal nerve tissue. Species crossreactivity was observed for the rat. Repeated absorption produced liver-specific antisera that reacted with antigenic sites distributed along the entire hepatocyte and sinusoidal cell surfaces. For the precise localization as well as the detection of low concentrations of both liver-specific and nonspecific antigens, the ultrastructural visualization of reactive sites proved essential.

Animals↗
PubMed1977

Localization of lipase-like immunoreactivity in porcine adipose, aortic and myocardial tissue.

Lipase has been purified from pig adipose tissue and antibodies have been produced in rabbit. By indirect immunofluorescence and immunoenzyme techniques lipase-like immunoreactivity was demonstrated in the intima of pig aorta, in the endothelial cells of the myocardium and in plasma membranes of adipocytes and skeletal muscle cells. Lipase activity in extracts of some of these tissues was inhibited by the addition of anti-lipase antibodies. At least part of the immunoreactivity in the examined tissues is due to active lipases.

Adipose Tissue↗
PubMed1976

Trophoblastic pseudotumor of the uterus: an exaggerated form of "syncytial endometritis" simulating a malignant tumor.

Twelve cases of a hitherto unrecognized pseudotumorous trophoblastic invasion of the myometrium are analyzed. Human chorionic gonadotropin (HCG) was identified by an immunoenzyme technique in the cytoplasm of the invasive cells. The lesion may be localized and only superficially invasive or deeply invasive and have a gross appearance suggesting a neoplasm in the excised uterus. In either case, this process has been confused with various types of malignant tumors, most often choriocarcinoma, from which it may be distinguished by an absence of the characteristic dimorphic population of cytotrophoblast and cyncytiotrophoblast. Most of the patients, who ranged in age from 19 to 41 years, presented with amenorrhea uterine enlargement and were thought to be pregnant, although only four of them had positive pregnancy tests. The most serious complication was uterine perforation, which occurred spontaneously in one patient and during curettage in five others. One patient died as a result, but the other 11 are alive without evidence of disase 6 months to 12 years after the diagnosis. Of the 11 surviving patients, four received no treatment other than curettage. Because this trophoblastic lesion is likely benign and simulates a malignant tumor on pathologic examination, the term "trophoblastic pseudotumor" has been chosen to designate it.

Adult↗
PubMed1978

Presence of common surface antigens(s) on endodermal tumors and embryonal tissues of rats, hamsters and mice.

Antiserum against yolk-sac carcinoma of rat was prepared in rabbits. After appropriate absorption in vitro or in vivo this antiserum was examined on different tumors and normal tissues or rat, hamster and mouse. The methods used were indirect immunofluorescence and indirect immunoperoxidase staining and cytotoxicity tests. The immune serum was found to react with the cell membrane of different rat and hamster yolk-sac carcinomas. It reacted also with the cell surface of rat hepatoma cells. By absorption on hyalin and blocking with amniotic fluid it was shown that the antigen was neither a basement membrane component nor alpha-fetoprotein. The antiserum was cytotoxic to yolk-sac carcinoma and hepatoma cells. The immune reaction was limited to the cell membrane, as observed in immunofluorescence and in immunoperoxidase staining. The specificity of the antiserum was proved by cross-absorptions with various tumor lines and by removing its activity with the soluble fraction of yolk-sac carcinoma cells. Non-endodermal rat and hamster tumor lines did not react with the anti-yolk-sac carcinoma immune serum. Most normal adult tissues, including spermatozoa, were negative, but a positive reaction was observed in ovaries and on glandular cells of the uterus. In embryonal tissues this surface antigen(s) was detected in the endoderm of 8-day-old rat embryos 7-day-old mouse embryos and in yolk-sac endoderm of both species. The data indicate that the antigen(s) is associated with endodermal differentiation.

Animals↗
PubMed1976

The thyrotropic cells of the guinea pig pituitary. Electron microscopic study after characterization by immunocytochemical means.

Three different immunocytoenzymatic techniques were used to identify and characterize the thyrotropic cells in the pituitary of normal guinea pigs at the ultrastructural level (superimposition technique, immunocytochemical technique using P.A.P. and indirect immunohistoenzymatic method before embedding). These cells are characterized by a dark cytoplasm with granules ranging from 1500 to 2000 A in diameter. The appearance of these granules is very variable: some display a marked electron density and are homogeneous but some have a less marked electron density with a more electron dense peripherally situated region. The TSH molecules are essentially confined to the granules but when the immunocytochemical reactions are carried out before embedding, positive staining is also seen in the cytoplasm and the outer surface of most of the rough endoplasmic reticulum membranes. These results are discussed.

Animals↗
PubMed1977

California encephalitis virus transmission by arctic and domestic mosquitoes.

A zero passage arctic mosquito isolate of California encephalitis (CE) virus (showshoe hare subtype) was transmitted by wild-caught Aedes communis mosquitoes after 13 days incubation at 13 degrees and 23 degrees C, AFTER 20 days incubation at 13 degrees C, when mosquitoes imbibed 1 mouse LD50 in a blood meal. Transmission occurred after 20 days incubation at 13 degrees and 23 degrees C when mosquitoes were injected intrathoracially with 1 or 0.1 mouse LD50. Virus was also transmitted by A. aegypti 13 days after infection with 100 mouse LD50 by feeding or intrathoracic injection, and incubation at 13 degrees C. Virus antigen was detected in salivary glands of 42 percent virus-positive A communis mosquitoes by direct immunofluorescence, and in 50 percent or less of A aegypti mosquitoes by immunoperoxidase and immunofluorescence, with somewhat greater regularity by the indirect than the direct technique.

Aedes↗
PubMed1978

Evaluation of an indirect fluorescent antibody test, enzyme-linked immunosorbent assay and quantification of immunoglobulins in the diagnosis of bovine trypanosomiasis.

An indirect fluorescent antibody test (IFAT), a microscale version of the enzyme-linked immunosorbent assay (microELISA) and determination of IgM levels in serum were assessed for their comparative diagnostic value in the detection of bovine trypanosomiasis. Serum samples from drug-treated N'dama cattle and untreated N'dama and Zebu cattle from Liberia were examined for the presence fo antibodies to trypanosomes. In the untreated Zebu cattle, infections with T. vivax predominated and the prevalence of infection was higher than that found in untreated N'damas in which infections with T. congolense predominated. The proportion of animals which showed serological evidence of trypanosomiasis in the untreated Zebus was slightly higher than that found in the untreated N'damas. The prevalence of infection was low in N'dama cattle which had been treated with diminazene aceturate and homidium chloride but 50% of the animals showed serological evidence of trypanosomiasis. More serologically positive animals were detected by microELISA than IFAT, but both tests were equally sensitive in detecting antibodies in cattle in which trypanosomes were demonstrated by examination of peripheral blood. With both IFAT and microELISA it was necessary to carry out tests using antigens prepared from T. brucei, T. vivax and T. congolense in order to detect all serologically positive animals. Increases in serum IgM occurred in both N'dama and Zebu cattle but the levels were raised in only approximately half of the known infected animals. Overall, more animals gave positive reactions with IFAT and microELISA than showed raised IgM levels.

Animals↗
PubMed1979

Enzyme immunoassay for feline oncornavirus-associated cell membrane antigen (FOCMA) and detection of FOCMA in cell extract by enzyme immunoassay inhibition test.

An enzyme immunoassay (EIA) for FOCMA has been developed. The assay uses alkaline phosphatase-conjugated rabbit anti-cat IgG as the second antibody and p-nitrophenyl phosphate as the substrate for the enzyme to detect cat FOCMA antibody bound to the target cells. In comparison with the indirect immunofluorescence (IIF) test, which was originally used for FOCMA assay, our results showed a good correlation between the two methods. The EIA gives a more objective measure of FOCMA reactivity than does IIF. FOCMA was successfully extracted from FOCMA-positive cell membranes by 0.5% Triton X-100 and further fractionated by ammonium sulfate. The FOCMA activity was assayed by IIF and EIA inhibition test. Most of the FOCMA activity was found in the fractions precipitated by 30% and 50% ammonium sulfate saturation.

Animals↗