Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0019163 (hepatitis B)
 38,309 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Optimal conditions were sought for the radiolabeling of microgram quantities of hepatitis B surface antigen (HBs Ag) employing the chloramine-T or lactoperoxidase iodination procedures. Preparations of HBsAg labeled by these procedures are referred to as chloramine-T preparations and lactoperoxidase preparations, respectively. Labeled HBsAg having specific activities between 10-20 muCi/mug were found to display the greatest degree of sensitivity for unlabeled HBsAg and for anti-HBs using a double-antibody radioimmunoassay (RIA-DA). Increasing the specific activity above this level redulted in a decreased affinity of labeled 1251-HBs Ag for anti-HBs, indicating that soluble antigenic alterations had developed. At equivalent specific activities, chloramine-T preparations competed less effectively for unlabeled HBs Ag than lactoperoxidase preparations, and anti-HBs endpoint titers were slightly reduced, especially among preparations of high specific activity (greater than or equal to 65 muCi/mug). Chloramine-T preparations of HBs Ag (sp. act. 15--30 muCi/mug) showed essentially no antigenic deterioration over a 2-month period at minus 196 degrees C or minus 70 degrees C. Utilization of optimally labeled 1251-HBs Ag has increased the sensitivity of the RIA-DA for unlabeled HBs Ag 30-fold to a level below 1 ng/ml and enhanced antiamine-T method revealed that only the most acidic population was labeled (pH 3.75+/-0.5). In contrast, six antigenic components with distinct pI values ranging from 3.7 to 5.2 were detected by RIA-DA in both unlabeled HBs ag and in the chloramine-T preparation. This indicated that the chloramine-T method did not radically change the relative number or charge of each of the pI populations present in purified preparations of HBs Ag. Analysis of HBs Ag iodinated by the lactoperoxidase procedure revealed the presence of three of four populations of particles with pI values ranging from 3.9 to 4.5, suggesting that this procedure labels HBs Ag more uniformly.
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PMID:Immunological and biophysical properties of hepatitis B antigen labeled by the chloramine-T and by the lactoperoxidase methods. 5 Mar 82

Hepatitis B antigen (HBAg) has been demonstrated in conventional formalin-fixed paraffin-embedded liver tissue by peroxidase and fluorescent immunostaining as well as by orcein. Complete locational and morphologic identity is seen between material stained by specific immunologic methods and by orcein. The antigen is restricted to the cytoplasm and is generally observed in the hepatocyte; it is present in three morphologic forms. Certain morphologic forms can even be identified in hematoxylin and eosin-stained tissue. Results of immunostaining procedures indicate that the antigen demonstrated in this study consists entirely of surface coat of hepatitis B virus (HBsAg). This seems to be the only component revealed by orcein staining. The latter is considered to be a good marker of the surface antigen and to have certain advantages over immunostaining. It is suggested that suitability of conventional paraffin sections for the detection of HBAg has wide and important implications.
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PMID:Localization of hepatitis B surface antigen in conventional paraffin sections of the liver. Comparison of immunofluorescence, immunoperoxidase, and orcein staining methods with regard to their specificity and reliability as antigen marker. 5 76

A sensitive method for demonstrating the site of hepatitis B surface antigen (HBsAg) in fixed tissues embedded in either paraffin or araldite is described. The method employs the peroxidase-rabbit antiperoxidase linkage through goat antirabbit to rabbit anti-HBsAg. In staining hepatitis antigen in agar, comparison of fixation (using three common fixatives) with unfixed precipitation arcs revealed no recognizable differences in antigenicity induced by fixation. The method allows confirmation of positive reaction by appropriate blocking controls. The technic is compared with the orcein stain of Shikata and found to be somewhat more sensitive but slightly more time-consuming.
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PMID:An immunoperoxidase technic for the demonstration of the hepatitis B surface antigen in human livers. 5 16

In recent studies extrahepatic manifestations of viral hepatitis have been recognized as immune complex diseases. Hepatitis B surface antigen (HBsAg) has been successfully identified in immune complexes, but the pathogenic role of HBsAg-containing immune complexes (IC) remains questionable. The subject of the present study was the antigen-specific determination of IC in the course of hepatitis B virus infection using a new HBsAg-specific IC test (Pernice & Sedlacek, 1978). This test is based on the following principle: rabbit anti-HBs-coated polystyrole test tubes are incubated with the IC-containing test sample. The HBsAg-containing IC bind to the solid phase by their free antigenic determinants. There they can be quantified using a peroxidase-labelled anti-human IgG antibody. A good correlation was found between the level of HBsAg-containing immune complexes and the clinical state of six patients in a follow-up study. IC could be detected simultaneously with HBsAg and either decreased or disappeared before the occurrence of free anti-HBs. In the sera of an additional twenty-eight patient suffering from chronic active hepatitis, HBsAg-containing immune complexes were detected in 85% of cases. One patient suffering from polyarteritis nodosa was also positive. Occasionally, extremely high levels of IC were found in the course of these diseases.
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PMID:Antigen-specific detection of HBsAG-containing immune complexes in the course of hepatitis B virus infection. 9 65

Hepatitis B core antigen (HBcAg) and hepatitis B surface antigen (HBsAg) were localized in human liver tissues by the peroxidase-labeled antibody method at the light and electron microscopic levels. Several methods of fixation, staining, and inhibition of endogenous peroxidase activity were studied. The periodate-lysine-paraformaldehyde fixative effectively preserved the tissue structure and the antigenicity of both antigens, and the peroxidase-labeled Fab' fraction of IgG penetrated well into hepatocytes. HBcAg was present in nuclei, or cytoplasm of hepatic cells, or both. In nuclei, the antigen was found both in virus-like particles of approximately 20 nm. diameter and in nuclear ground substance. In the cytoplasm, the antigen was found on membrane-bound ribosomes and free polysomes, and also in the ground substance of the cytosol near ribosomes and around nuclear membranes, especially near nuclear pores. HBcAg-positive virus-like particles were also demonstrated sparsely or in clusters in the cytoplasm. HBsAg was not present in nuclei but was found in the perinuclear space and in cisternae of endoplasmic reticulum, and on nuclear, endoplasmic reticulum, and cell membranes of hepatic cells. HBsAg-positive 25- to 30-nm. wide tubular forms, round particles (probably cross-sections of tubular forms), and a few large particles of 40 to 50 nm. diameter were seen in cisternae. Such HBsAg-positive particles were also present in the intercellular space and in Disse's space. These findings suggest that HBcAg produced on the cytoplasmic ribosomes migrates through nuclear pores to the nucleus and is assembled into core particles there. These particles may then move through nuclear pores to the cytoplasm where they are invested with HBsAg-positive membrane in cisternae of endoplasmic reticulum or as they enter the endoplasmic reticulum. These virus particles are then released together with other HBsAg-positive forms into the intercellular space by reversed phagocytosis.
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PMID:Hepatitis B core and surface antigens in liver tissue. Light and electron microscopic localization by the peroxidase-labeled antibody method. 32 96

A solid-phase enzyme immunoassay for the qualitative detection of hepatitis B surface antigenemia is presented and the results compared to the counter-immunoelectrophoresis and radioimmunoassay methods for detection of the same antigen. The enzyme-antibody conjugate was prepared from horseradish peroxidase coupled with antibodies to three sub-types of hepatitis B virus. Polystyrene plastic tubes were used as the solid-phase support. The results of the enzyme immunoassay compare favorably with radioimmunoassay and exceed counterimmunoelectrophoresis in sensitivity.
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PMID:Enzyme immunoassay for hepatitis B and its comparison to other methods. 33 44

IgM antibody against core antigen of the hepatitis B virus (anti-HBc IgM) was selectively determined by a new enzyme immunoassay (EIA). Microtiter plates were coated with anti-human micro chain immunoglobulin. On addition of serum IgM is bound by a factor of about 4,000 more than IgG. After removing the sample, HBcAg is added to the IgM-coated surface. Binding takes place if the IgM contained anti-HBc and was demonstrated by the aid of a conjugate made from anti-HBc IgG and horse radish peroxidase. Quantitation may be achieved without testing a dilution series. The assay was not disturbed by a large excess of anti-HBc IgG in the sample and rheumatoid factor did not produce false-positive results, provided the sample was diluted in an excess of aggregated IgG. The diagnostic relevance of the assay was demonstrated in selected cases of acute hepatitis B. Rapid diagnosis of acute hepatitis B infection is therefore now possible in those cases whihc are HBsAg-negative but anti-HBc-positive.
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PMID:Selective detection of IgM-antibody against core antigen of the hepatitis B virus by a modified enzyme immune assay. 39 75

The present report describes a sensitive and reliable method for potentiometric determination of hepatitis B surface antigen in biological fluids, as an application of enzyme-linked immunoassay technique by a "sandwich" procedure. Specific antibodies are immobilized on a gelatine membrane which covers the sensor of an iodide-sensitive electrode. After immersion of the modified electrode in a dilute solution of antigen, the enzymatic activity is evaluated in a solution of peroxidase-labelled antibodies, in the presence of substrate and iodides. The electrode potential is a function of antigen concentration in the solution. This new procedure should find its application in the determination of substances present in very low concentration in biological fluids.
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PMID:[Potentiometric determination of hepatitis B surface antigen in biological fluids (author's transl)]. 69 28

Purified 22-nm forms of hepatitis B surface antigen (Hbsag) representing the three major antigenic subtypes (adw, ayw, and adr) were analyzed for their constituent polypeptides by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. No consistent difference in either the number or relative distributions of the polypeptides was observed for the various subtypes. Seven polypeptides were designated as P-1 through P-7 in order of their decreasing mobilities. By comparison with protein standards, their molecular weights were estimated as 23, 29.5, 36, 41.5, 53.5, 72, and 97 thousand. The P-1 and P-2 components represented the major polypeptides; P-2 and P-5 might by glycoproteins, based on their reaction with periodic acid-Shiff reagent. Each polypeptide contains cysteine residues. HBSAg was radiolabeled with 3H or 14C by reductive methylation or iodinated with 125I by the chloramine-T or lactoperoxidase procedures. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of labeled HBSAg yielded patterns identical to those obtained with protein stain. Comparison of HBSAg labeled by the chloramine-T and lactoperoxide procedures indicated that there was no distinction between internal or external components within the 22-nm structure.
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PMID:Proteins of hepatitis B surface antigen. 83 27

Peritoneal liver biopsy specimens from eight patients with hepatitis B associated cirrhosis, complicated by hepatocellular carcinoma, were studied for identification and localisation of myofibroblasts. The avidin-biotin peroxidase complex technique was used on paraffin wax sections, using monoclonal antibodies for actin and desmin, and ultrastructural examination was performed. Myofibroblasts were found in seven of the eight cirrhotic specimens and in all eight tumour specimens. They were identified in the fibrotic areas by the immunohistochemical technique, but ultrastructural examination disclosed their presence in the perisinusoidal space and between tumour cells.
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PMID:Myofibroblasts in hepatitis B related cirrhosis and hepatocellular carcinoma. 131 87


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