Atsushi Shimoda

Expression of interferon alpha/beta receptor in the liver of chronic hepatitis C patients

Interferon (IFN) demonstrates antiviral activity by binding to receptors on the cell surface. Expression of the IFN receptor in hepatocytes may be directly associated with a hepatitis C virus (HCV) infection and the response to IFN therapy. A competitive PCR method was developed to measure IFN alpha/beta (α/β) receptor mRNA in liver samples obtained by needle biopsy. Thirty‐one patients with chronic hepatitis C (21 without cirrhosis, 10 with cirrhosis) and six normal subjects were used. Eighteen of the 21 patients without cirrhosis received the IFN therapy. Competitive PCR was carried out using IFN α/β receptor gene‐specific primers and a specific competitor. Expression of the receptor was detected in all liver samples. There was no association between the expression level and serum alanine aminotransferase level, serum (2′–5′) oligo (A) synthetase level, amount of serum HCV RNA, or HCV genotype. The expression level in patients with chronic hepatitis was significantly higher than that in normal livers (P < 0.05) and in cirrhotic livers (P < 0.01). Seven of the 18 patients treated with IFN demonstrated a sustained response to IFN (sustained responders), and the remaining 11 did not (nonsustained responders). The expression level of IFN α/β receptor mRNA in the sustained responders was significantly higher than that in the nonsustained responders (P < 0.01). Thus, the expression of IFN α/β receptor mRNA may be one of the host factors influencing the response to IFN therapy. J. Med. Virol. 56:217–223, 1998.

Gastrointestinal amyloidosis secondary to hypersensitivity vasculitis presenting with intestinal pseudoobstruction

Hypersensitivity vasculitis is characterized byinflammation and necrosis of small blood vesselssecondary to allergic or hypersensitivity mechanisms (1,2). Gastrointestinal involvement with edema and bleeding also has been reported (3-5).Long-standing inflammation, such as rheumatic disease,infectious disease, inflammatory bowel disease, familialMediterranean fever, and malignancy, may lead tosystemic amyloidosis (6). Gastrointestinal involvementmay induce anorexia, nausea and vomiting, diarrhea,constipation, bleeding, malabsorption, andpseudoobstruction (6, 10-12). In this report we discussa patient with hypersensitivity vasculitis with severeintestinal bleeding who developed systemic amyloidosiswith intestinal pseudoobstruction 29 months after onset.Secondary amyloidosis due to hypersensitivity vasculitis has not been previously reported,and the causal relationship is discussed in thisreport.

Interferon-alpha modulates resistance to cisplatin in three human hepatoma cell lines

Abstract: We investigated the expression of the drug resistance-related genes, multidrug resistance gene 1 (MDR1), multidrug resistance associated protein gene (MRP), and the DNA topoisomerase IIα, DNA topoisomerase IIβ, and glutathione-S-transferase π gene (GST-π) in three human hepatoma cell lines (HepG 2, HuH 7, SK-Hep-1) with or without drug treatment with interferon-α (IFN-α) and cisplatin (CDDP), by a reverse transcription-polymerase chain reaction (RT-PCR) method and a competitive PCR method. The signals of the MDR1, MRP, topoisomerase IIα, and topoisomerase IIβ genes in HepG2 were weakened when IFN-α was added to CDDP. In SK-Hep-1, the administration of CDDP alone increased the signals of MDR1 while the addition of IFN-α decreased the signals, and the signals of GST-π were decreased by IFN-α plus CDDP. In summary, our results concerning the expression of drug resistance-related genes in three human hepatoma cell lines demonstrate that IFN-α may modulate the mechanism of resistance to CDDP in liver cancer.

Hepadnavirus enhancer and its binding proteins

SummaryIn the hepadnavirus enhancer region, a 33 bp DNA sequence is strongly conserved among mammalian hepadnavirus genomes. To elucidate the role of the sequence, we tested enhancer activities and capability to form DNA-protein complex of several synthetic DNAs. Not only two tandem copies of a 46 bp DNA covering the sequence but also two tandem copies of a 23 bp in the sequence exhibit enhancer activity. Also the activity was augmented by treatment of a tumor promoter, TPA. DNA binding proteins complexes with the 23 bp DNA were augmented in extracts of HepG2 or HeLa cells stimulated with TPA. These results imply that the conserved sequence of hepadnavirus enhancer is a TPA-inducible enhancer which is transactivated by ubiquitous DNA-binding proteins. We presented results showing that DNA-protein complexes with a 23 bp DNA are similar to but distinct from those with a TPA-responsive element DNA, the recognition site for c-jun/fos products. We also presented results suggesting that hepadnavirus X protein may not directly or indirectly affect DNA-protein complex formation with the conserved sequence in the hepadnavirus enhancer.

A chimeric CYP11B1/CYP11B2 gene in glucocorticoid-insuppressible familial hyperaldosteronism

Although a chimeric gene combining the 11β‐hydroxylase gene (CYP11B1) and the aldosterone synthase gene (CYP11B2) explains the pathophysiology of familial hyperaldosteronism (FH) type I, the contribution of this abnormality to FH type II has not been tested. We screened genomic DNA from a Japanese family with FH type II for the CYP11B1/CYP11B2 gene. The index patient was a 27‐year‐old woman with hypertension. Hypokalaemia, elevated plasma aldosterone and suppressed plasma renin activity suggested primary aldosteronism. Though computed tomography failed to reveal an adrenal tumour, left adrenalectomy was indicated due to a high aldosterone concentration in left adrenal venous blood. The resected adrenal gland contained an adenoma. As her mother had also been diagnosed with primary aldosteronism due to an adenoma, we administered oral dexamethasone to our patient before the operation and observed the response of the blood pressure and plasma aldosterone concentration for 2 weeks. Both parameters remained elevated during the treatment period, confirming the diagnosis of FH type II. Total DNA was isolated from blood cells of the index patient, her mother, and an unaffected brother. Samples were amplified by polymerase chain reaction using specific primers from CYP11B1 and CYP11B2. Unique DNA fragments of 1·4 kb were obtained from the index patient and her mother, but not from the healthy subject. The CYP11B1/CYP11B2 chimeric gene was found in a Japanese family with FH type II.

A case of fulminant hepatitis subacute form detected variant hepatitis G virus.

症例は67歳, 女性. 平成8年9月初旬より全身倦怠感が出現し, 肝障害を指摘され増悪したため平成8年10月23日当科入院. 非A非B非C型劇症肝炎亜急性型と診断され, 血漿交換, インターフェロン投与などを行うも効なく11月19日肝不全にて死亡. 剖検では亜広範性肝壊死の所見であった. 本例の10月27日の血清を用いてG型肝炎ウイルスRNAの検索を4種類の異なるprimerによるPCR法で行ったところ2種類で陰性, 2種類で陽性と検出結果が異なった. そこで本ウイルスの全塩基配列を決定したところプロトタイプウイルスと大きく異なりホモロジーは約80%であることが判明した. G型肝炎ウイルスRNAが検出されなかった測定法のprimer部分で塩基配列が異なっており, PCR反応が起こらずRNAが検出されなかったものと思われた. 本例で検出されたG型肝炎ウイルスはこれまでのタイプとは異なる第4のサブタイプである可能性が考えられた.

Hepatitis G virus infection in Japanese patients with chronic liver disease

The clinical significance of anti-G in alloimmunized pregnant women We read with interest the recent article by Shirey et al.’ in which they describe three women whose sera seemed to contain anti-C and anti-D but which actually contained anti-C and anti-G. We agree with the authors’ contention that it can be important to identify the actual specificities of antibodies when a history of Rh prophylaxis or the presence of relatively high levels of anti-C suggests that anti-D may not actually be present. The authors point out that women with anti-C and anti-G (rather than anti-D) should continue to receive Rh prophylaxis. However, we are less comfortable with their statement that anti-G poses “less of a threat to the fetus than would anti-D or anti-C.” The rarity with which anti-G has been associated with severe hemolytic disease of the newborn presumably relates in’large part to the difficulty in distinguishing the relative contributions of anti-D and anti-G to the hemolytic processes in D-positive fetuses. The clinical significance of antiG may become apparent only when the fetus is D-negative. We have recently reported a case of severe hemolytic disease of the newborn due to anti-G in a D-negative (fr) fetus.2 Using the chemiluminescence test to indicate potential clinical significance: we showed that 2 of 28 maternal sera containing over 5 IU per mL of anti-C and anti-D actually contained levels of anti-G that were consistent with moderate to severe hemolytic disease of the newborn. Thus, we would add to the authors’ list of instances in which anti-G should be distinguished from anti-D those cases in which the fetus is D-negative and the mother apparently has antiC and anti-D. Andrew Hadley, DPhil Joyce Poole, FIBMS International Blood Group Referencekboratory Geoff Poole, BSc, MSc, FIBMS National Blood Service Southmead Road Bristol BS10 5ND, UK