Immunohistochemical investigation of hepatitis B virus associated antigens,HLA antigens and lymphocyte subsets in type B chronic hepatitis

HLA antigens, hepatitis B virus (HBV)-assodated antigens and lymphocyte subsets in liver tissue from 35 patients with HBs antigenemia were studied using an immunoperoxidase double staining method arid immunoelectron microscopy in order to clarify the immune mechanism of hepatocyte lysis in type B hepatitis. Immune light and electron microscopy using monoclonal antibodies to lymphocyte subsets revealed that infiltrating lymphocytes in the areas of piecemeal necrosis and focal necrosis were predominantly CD8-positive, showing direct contact with hepatocytes. In contrast, CD4(+) cells were infrequently observed in necrotizing inflammatory lesions. HLA-A,B,C antigens were mainly found on hepatocytes in areas of piecemeal necrosis and focal necrosis, in association with CD8(+) lymphocyte infiltration. HLA-DR antigens were demonstrated on a few hepatocytes in the same lesions. In cases of CAH with serum HBeAg positive, HLA-A,B,C, antigens and HBV antigens simultaneously demonstrated on the same hepatocytes. Especially, hepatocytes expressing both HLA-A,B,C antigen and HBsAg on the plasma membrane showed direct contact with CD8(+) lymphocytes. This finding fullfilled the morphological requirements for HBsAg as a target antigen. On the other hand, HBcAg was hardly demonstrated in the liver cell membrane but was demonstrated mainly in the cytoplasm. Compared with the nuclear localization of HBcAg in cases of NSR, cytoplasmic localization of this antigen may be associated with membranous expression of new antigens induced by HBV infection.     

Distamycin A modulates the sequence specificity of DNA alkylation by duocarmycin A

Duocarmycin A (Duo) normally alkylates adenine N3 at the 3′ end of A+T-rich sequences in DNA. The efficient adenine alkylation by Duo is achieved by its monomeric binding to the DNA minor groove. The addition of another minor groove binder, distamycin A (Dist), dramatically modulates the site of DNA alkylation by Duo, and the alkylation switches preferentially to G residues in G+C-rich sequences. HPLC product analysis using oligonucleotides revealed a highly efficient G–N3 alkylation via the cooperative binding of a heterodimer between Duo and Dist to the minor groove. The three-dimensional structure of the ternary alkylated complex of Duo/Dist/d(CAGGTGGT)·d(ACCACCTG) has been determined by nuclear Overhauser effect (NOE)-restrained refinement using 750 MHz two-dimensional NOE spectroscopy data. The refined NMR structure fully explains the sequence requirement of such modulated alkylations. This is the first demonstration of Duo DNA alkylation through cooperative binding with another structurally different natural product, and it suggests a promising new way to alter or modify the DNA alkylation selectivity in a predictable manner.     

Correction

The online version of the original article can be found at     

A New Platelet-Specific Antigen, Naka, Involved in the Refractoriness of HLA-Matched Platelet Transfusion

Serum from a thrombocytopenic patient who was refractory to the transfusions of HLA-matched platelets contained a platelet-specific alloantibody, anti-Naka. Immunofluorescence analyses revealed that the Naka antigen defined by the serum was expressed exclusively on platelets and its distribution was different from P1A1, Baka, Yuka or Yukb. Analysis by Dr. von dem Borne's group revealed the Naka was also different from Koa, Kob or Zwb. Family studies showed that the Naka antigen was inherited as an autosomal codominant trait. Its antigen frequency in the Japanese population was over 97%. The results of the enzyme immunoassay using monoclonal antibodies for antigen immobilization showed that the Naka epitope did not appear to reside on GPIIb/IIIa or Ib. The transfusions of Naka-compatible platelets improved the patient's thrombocytopenia.     

A case of Mycobacterium avium pulmonary disease detected as multiple small nodular shadows and diagnosed by CT guided transbronchial biopsy with ultrathin bronchoscopy]

A 65-year-old man was admitted to our hospital because multiple small nodular shadows measuring five to 12 mm in diameter in both lung fields were noticed on chest radiographs and CT scans during a regular checkup for pulmonary emphysema. The levels of ESR and CRP were within normal limits, but those of CEA and CYFRA were slightly elevated. Mycobacterium avium (M. avium) was detected in sputum cultures and in a sputum examination using PCR. Non-necrotizing granulomas were detected in the specimens obtained from a lesion of the right S6b by CT-guided transbronchial biopsy using an ultrathin bronchoscope after navigation with virtual bronchoscopy. On the basis of these findings, M. avium pulmonary disease was diagnosed. The patient is being followed up, but because he has no symptoms, without medication. To our knowledge, there have been no previous reports of M. avium-intracellulare complex pulmonary disease, which is characterized by multiple small nodular shadows. It is speculated that multiple centrilobular lesions developed simultaneously without extension to the bronchial walls, leading in this case to multiple small nodules in both lung fields.     

Histone deacetylase inhibitors stimulate cell migration in human endometrial adenocarcinoma cells through up-regulation of glycodelin

Histone deacetylase inhibitors (HDACIs) have recently emerged as promising anticancer drugs to induce cell cycle arrest, cytodifferentiation, and apoptosis. It is suggested, however, that HDACIs promote cell migration and invasion depending on the cell type. We have reported previously that treatment with HDACIs, including trichostatin A and suberoylanilide hydroxamic acid (SAHA) or progesterone in combination with estrogen, can induce cytodifferentiation of endometrial adenocarcinoma Ishikawa cells through up-regulation of glycodelin, a progesterone-induced endometrial glycoprotein. Given the reported role of glycodelin in cell motility and the migration-modulating potential of HDACIs, we investigated using wound healing assay and transwell migration assay whether ovarian steroid hormones, trichostatin A, or SAHA affects cell migration in endometrial cancer cell lines, Ishikawa and RL95-2. Treatment with ovarian steroid hormones, trichostatin A, and SAHA enhanced cell migration together with up-regulation of glycodelin. SAHA-augmented cell migration was almost completely blocked by gene silencing of glycodelin. Furthermore, overexpression of gycodelin alone resulted in increased cell motility in Ishikawa cells. Our results collectively indicate that glycodelin positively regulates cell motility acting as a mediator of HDACI-enhanced endometrial cell migration, suggesting the involvement of glycodelin in the dynamic endometrial gland morphogenesis during menstrual cycle. Our results raise a possibility that the use of HDACIs in the therapy for glycodelin-inducible endometrial and presumably other gynecological cancers may enhance invasion in cases in which the HDACIs fail to exert differentiation-inducing and/or antiproliferative effects.     

Clinical features of hepatitis C virus-related hepatocellular carcinoma and their association with alpha-fetoprotein and protein induced by vitamin K absence or antagonist-II.

PURPOSE: We investigated the differences in clinical features between alpha-fetoprotein (AFP)-predominant hepatocellular carcinoma (HCC) and protein induced by vitamin K absence or antagonist-II (PIVKA-II)-predominant HCC, especially regarding host factors thought to contribute to hepatocarcinogenesis in chronic hepatitis C virus (HCV) infection. METHODS: HCV-related HCC patients (n=306) were divided into four groups according to median AFP (48.1 ng/ml) and PIVKA-II (60 mAU/ml). Host factors, tumor factors, survival, and risk factors affecting survival were compared. RESULTS: Aspartate aminotransferase (AST; IU/L), alanine aminotransferase (ALT; IU/L), and platelet count (x 10(4)/ml) were, respectively, 81, 67, and 8.2 in AFP-predominant HCC (group A; n=66) vs. 50, 42, and 11.4 in PIVKA-II-predominant HCC (group P; n=52). Tumor sizes (mm) in groups A and P were 20 and 37, respectively. Significant differences were evident. Survival was identical between the two groups. Factors affecting survival were total bilirubin, portal tumor thrombus and number of nodule in group A, and albumin and tumor distribution in group P. CONCLUSIONS: PIVKA-II-predominant HCC had a milder hepatitis and a better-preserved platelet count compared with AFP-predominant HCC. Considering the strong relation between hepatocarcinogenesis and hepatic inflammation with chronic HCV infection, these differences indicate that hepatocarcinogenic mechanisms in PIVKA-II-predominant HCC may differ from those in AFP-predominant HCC.