Kiyoshi Ishihara

Autoantibodies against a 210kDa glycoprotein of the nuclear pore complex as a prognostic marker in patients with primary biliary cirrhosis

It has been reported that the presence of anti‐nuclear antibody against a 210kDa glycoprotein of nuclear pore complex (anti‐gp210) is highly specific for primary biliary cirrhosis (PBC). The aim of the present study was to investigate the significance of anti‐gp210, especially as a prognostic marker. The presence of anti‐gp210 was ascertained in 113 patients with PBC and 162 controls by indirect immunofluorescence assay using HepG2 cells and immunoblotting analysis using nuclear extracts from HeLa cells. Anti‐gp210 was detected in 25 of the 113 (22.1%) patients. None of the 162 controls was positive for anti‐gp210. The appearance and titre of anti‐gp210 in the patients with PBC did not vary from the time of diagnosis and through their clinical course. Anti‐mitochondrial antibodies (AMA), including antibodies against pyruvate dehydrogenase complex, branched chain α‐ketoacid dehydrogenase complex and α‐ketoglutarate dehydrogenase complex, were not detected by enzyme‐linked immunosorbent assay in five of the 113 (4.4%) patients with PBC. However, anti‐gp210 alone was positive in one of these five patients. The difference in prognosis was statistically significant; patients with PBC positive for anti‐gp210 died from hepatic failure more frequently than those who were negative (P < 0.01), although there were no statistically significant differences in the frequency of jaundice and the histological stage at the time of diagnosis between the two groups. We suggest that the presence of anti‐gp210 is one of the independent prognostic markers able to predict, at the time of diagnosis, a poor outcome in patients with PBC.

Hepatic lesions induced by graft-versus-host reaction across MHC class II antigens: An implication for animal model of primary biliary cirrhosis

To induce graft-versus-host reaction (GVHR), C57B1/6 (B6) spleen cells were injected into (B6 x bm1)F1, (B6 x bm12)F1, and (bm1 x bm12)F1 mice. Since the strains bm1 and bm12 are mutant at the H-2Kb and I-Ab regions of major histocompatibility complex (MHC), respectively, we can assess MHC class I- or class II-different GVHR. As reported earlier, immunological perturbations assessed by the number of immunoglobulin-producing cells and immune complex deposition in renal glomeruli were demonstrated in MHC class II-different GVHR. A conspicuous finding in this report is that epithelioid granuloma formation was observed in the portal area and around the central vein of liver of (B6 x bm12)F1 mice injected with B6 spleen cells. The epithelioid granuloma formation was not observed in (B6 x bm1)F1 nor (bm1 x bm12)F1 recipient mice. Degenerative changes resembling chronic nonsuppurative destructive cholangitis in primary biliary cirrhosis were also observed in the bile duct epithelium in (B6 x bm12)F1 and (bm1 x bm12)F1 mice. These lesions were already obvious at the 2 week postinjection of donor cells and were continuously observed up to 10 weeks when immunological perturbations subsided. Thus, class II-disparate GVHR in this experimental system might provide a novel animal model of protracted disease, primary biliary cirrhosis.

Remote development of hepatocellular carcinoma in patients with liver cirrhosis type B serologically cured for HBs antigenemia with long-standing normalization of ALT values.

In this report, we examine two patients with chronic hepatitis B virus (HBV) infection that had been diagnosed as precirrhosis or liver cirrhosis more than a decade previously. These patients had been cleared of HBsAg and had developed anti-HBs at a later time, yet hepatocellular carcinoma (HCC) eventually occurred. Both patients had been found negative for HBV DNA, using sensitive methods. Interestingly, a nontumor specimen of the liver obtained at surgical resection showed a marked reduction of fibrosis when compared to the histology observed when the patient was diagnosed as precirrhosis. Our findings suggest that the fibrosis from liver cirrhosis had been absorbed to a large extent during the long-term absence of active viremia and the normalization of alanine aminotransferase (ALT) levels. However, the cancer-prone biological characteristics of liver cirrhosis remained. Thus, patients with liver cirrhosis due to past chronic hepatitis B should be monitored carefully for the development of HCC even if HBV infection has been serologically resolved.

Treatment of symptomatic primary biliary cirrhosis with lymphocytapheresis: a preliminary clinical trial

Abstract Lymphocytapheresis was carried out as a preliminary clinical trial for four patients with symptomatic primary biliary cirrhosis (PBC). After lymphocytapheresis, severe itching disappeared in two patients and jaundice improved or disappeared in all cases. Serum levels of total bilirubin, alkaline phosphatase, γ -glutamyl transpeptidase and transaminase were decreased after the treatment. In liver tissue, the number of infiltrated lymphocytes was slightly decreased in both the parenchyma and portal tract. Increased CD3 − CD56 + lymphocytes (NK cells) in the liver were dramatically diminished after the treatment. CD3 + CD56 − lymphocytes (T cells), especially CD8 + lymphocytes and CD56 + T lymphocytes, were increased after the treatment in two cases. Although these results are preliminary and based on only a small number of patients and over short observation periods, lymphocytapheresis might be a useful therapeutic tool for patients with advanced PBC, giving excellent improvement of symptoms and results of liver function tests.

Amplification of BCR Protein Associated with Oncogenesis in Human Hepatocellular Carcinoma

The BCR gene is located on human chromosome 22.The normal cellular BCR gene encodes a 160,000-daltonphosphoprotein associated with a serine/threonine kinaseactivity. The BCR protein is involved in signal transduction. We investigated the expression ofthe BCR protein in hepatocellular carcinoma (HCC),surrounding noncancerous liver tissue, liver cirrhosis(LC), chronic hepatitis (CH), and normal liver with immunohistochemistry and a western blotanalysis. BCR immunoreactivity was detected using amonoclonal antibody. In normal liver, and both CH and LCwithout association of HCC, the immunoreactivity of the BCR protein was minimal. In contrast, 73% (22of 30) of noncancerous liver tissue adjacent to the HCCand 40% (12 of 30) of HCC expressed BCR protein; thisdifference was statistically significant (P < 0.01). The expression of the BCR protein expressioncorrelated with the degree of histologicaldifferentiation of HCC (P < 0.05). In addition, theamplification of BCR protein in noncancerous cells wassupported by the detection of specific protein using awestern blot analysis. In two cases, the expression ofBCR protein occurred only in overtly malignant HCCcells. As a result, the expression of the BCR protein may be associated with oncogenesis in humanHCC.

Abnormality of serum immunoglobulins in chronic liver disease-immune complex like substances, abnormaly basic γ-globulin, secretory IgA and anti-DNA antibody

1) Serum secretory IgA was reduced in chronic liver disease while it was increased in obstructive jaundice. 2) Serum anti-dsDNA antibody was slightly increased in chronic liver disease, especially it was significantly increased in lupoid hepatitis in which its titer paralelled with the course of disease activity. 3) Serum Clq binding activity, Clq binding inhibition activity and polyclonal rheumatoid factor binding inhibition activity were increased in chronic liver disease and their disease activity was correlated with concentration of macromolecular immune complexes which were fractionated with sucrose density gradient ultracentrifugation. 4) Abnormally basic y-globulin which was dominantly found in chronic hepatitis B sera was determined to be monomeric IgG. I t was increased in aggravation of the disease but has no correlation with Clq binding monomeric IgG. 5) Liver membrane specific lipoprotein (LP-1), Espinosas liver specific antigen (LSA), Nerenbergs hepatorenal antigen (HRA) and Tamm-Horsfall glycoprotein (THGP) had positive charge, and LP-2 and F-antigen did negative charge. 6) Human liver cell membrane fraction could not be obtained by the method of Ray or aqueous two phase polymer system which have been used for rat liver.