Orcein staining of hepatitis B antigen in paraffin sections of liver biopsies.

Abstract

SYNOPSIS Liver biopsies from 97 hepatitis B antigen (HB%Ag)-positive patients were stained by a modified orcein method described by Shikata et al (1974) Although more than 10 years have elapsed since the discovery of hepatitis B antigen (HBsAg) (Blumberg, 1964), its localization in tissues has hitherto remained a complicated procedure, requiring immunofluorescence or electron microscopy. Recently, however, Shikata, Uzava, Yoshiwara, Akatsuka, and Yamazaki (1974) have developed simple histochemical methods for the demonstration of HB.Ag in conventional paraffin sections. The mechanism is apparently based on the presence of disulphide bonds in HBsAg (Vyas, Rao, and Ibrahim, 1972a;Sukeno, Shirachi, Yamaguchi, and Ishida, 1972). This paper reports the results of staining by one method described by Shikata et al (1974) in liver biopsies from patients with acute and chronic liver disease and from asymptomatic carriers of the antigen. Material and MethodsTen of the patients who underwent liver biopsy were from the Withington Hospital, Manchester, and the remainder from the Royal Free Hospital, London.The liver biopsies studied, 12 of them operative wedge biopsies and the remainder percutaneous needle biopsies, were from 148 patients whose overall clinico-pathological diagnoses are shown in table I.

Detection of HBSAG in fixed liver tissue - use of a modified immunofluorescent technique and comparison with histochemical methods.

“…Furthermore, positive hepatocytes were not clumped together, but were scattered in random fashion throughout each lobule, although frequently, where the total number of positive cells was small, some lobules were negative. We were unable to confirm the previous observation (Shikata et al, 1974;Deodhar et al, 1975) inverse relationship between cells containing HBsAg and areas of piecemeal necrosis (Fig. 3).…”

“…An inverse relationship at a morphological level in chronic liver disease between the presence of orcein-positive hepatocytes and areas of necrosis was reported both by Shikata et al (1974) and by Deodhar et al (1975). A similar distribution was noted previously using immunofluorescent techniques in two other studies (Hadziyannis et al, 1972;Shikata, 1973).…”

“…It now seems likely that the antisera employed in the earlier immunofluorescent studies also contained antibodies directed at the hepatitis B core-antigen (HBcAg) or against nuclear proteins (Gerber and Paronetto, 1974). Moreover, nuclear staining has never been observed with orcein (Shikata et al, 1974;Deodhar et al, 1975). The quantity of HBsAg in the cytoplasm varied somewhat from cell to cell and in some, was distributed diffusely, while in others, the pattern was more discrete.…”

Investigation of korean plant extracts for potential phytotherapeutic agents against B‐virus hepatitis

“…The precise mode(s) of viral pathogenesis remains unknown (Phillippe and Joseph, 1981 ;London and Joseph, 1985). Epidemiological studies predict that a large number of patients actively infected with HBV will develop liver cirrhosis, and approximately 35% of this group will progress to primary hepatocellular carcinoma (Deodhar et al, 1975;Sakurai and Miyaji, 1977).…”

New Insights into the Neuropathogenesis of Molybdenum Cofactor Deficiency

“…The mechanism of staining is related to the presence of disulphide (S-S) bonds. 9,10 We have used these stains on brain specimen at our institution. We did not find any published report in the literature on their use in brain sections hence an age-matched control case was used in our study.…”