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Detection of Epstein-Barr virus in Hodgkin-Reed-Sernberg cells -: No evidence for the persistence of integrated viral fragments in latent membrane protein-1 (LMP-1)-negative classical Hodgkin's disease

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AMERICAN JOURNAL OF PATHOLOGY 2000年第1期156卷 209-216页

作者： Staratschek-Jox, A Kotkowski, S Belge, G Rüdiger, T Bullerdiek, J Diehl, V Wolf, J Univ Cologne Dept Internal Med 1 D-50924 Cologne Germany Univ Bremen Ctr Human Genet & Genet Counseling Bremen Germany Univ Wurzburg Inst Pathol Wurzburg Germany

Classical Hodgkin's disease (HD) is associated with Epstein-Barr virus (EBV) infection. Although in developing countries EBV can be demonstrated in Hodgkin-Reed-Sternberg (H-RS) cells in up to 95% of HD cases, in industrialized countries only about 50% of HD cases are associated with EBV. An open question remains whether EBV in the EBV-negative cases has escaped detection by standard screening procedures due to deletions in the viral genome associated with integration of viral fragments into the host cell genome. We, among others, recently described this phenomenon in Burkitt's lymphoma cells. To investigate whether H-RS cells in latent membrane protein-1 (LMP-1)-negative HD cases harbor fragments of the EBV genome, we combined fluorescence in situ hybridization (FISH) using a set of six overlapping DNA probes spanning the whole EBV genome with immunophenotyping of fresh frozen lymphoma sections. Results in the eight cases analyzed were as follows: in three LMP-1-positive cases, FISH analysis yielded specific signals for each EBV DNA probe in H-RS cells, which had been identified by morphology and CD30 staining. In contrast, none of the EBV DNA probes hybridized to the H-RS cells in the five LMP-1-negative cases, Thus, there is no evidence for the presence of fragments of the viral genome integrated into the host cell genome in the LMP-1-negative cases. Furthermore, in the LMP-1-positive cases analyzed, no large deletions in the viral genome were detected. These results show that, in classical HD, LMP-1-negative cases do not harbor EBV DNA within the H-RS cells. Whether, in these cases, a still unknown virus contributes to the transformation and maintenance of the malignant phenotype remains to be established.

关键词： DNA 病毒/分析基因组病毒疱疹病毒4型人/遗传学疱疹病毒4型人/分离和提纯霍奇金病/代谢霍奇金病/病理学霍奇金病/病毒学免疫表型分型原位杂交荧光里德-斯特恩伯格细胞/病毒学病毒基质蛋白质类/代谢病毒粒子/分离和提纯青少年成年人女(雌)性人类男(雄)性中年人

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Two types of HTLV-1 particles are released from MT-2 cells

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VIROLOGY 1999年第2期255卷 279-284页

作者： Morozov, VA Weiss, RA Inst Canc Res Chester Beatty Labs London SW3 6JB England Russian Acad Med Sci Canc Res Ctr Inst Carcinogenesis Moscow 115478 Russia

The MT-2 cell line transformed by human T-cell leukemia virus type 1 (HTLV-1) contains one complete provirus and seven defective proviruses. Four defective genomes have an identical structure (LTR-MA-delta CA-pX-LTR) with an open reading frame that spans from MA to pX, giving rise to a 3.4-kb (24S) RNA transcript encoding a chimeric Gag-pX protein, p28. MT-2 cells release two distinct types of virions. The major "classic" type of particle has a buoyant density of 1.155-1.16 g/cm(3) and contains the standard HTLV-I structural proteins and reverse transcriptase (RT). In addition, about 5% of particles are "light" approximately 1.12 g/cm(3), and contain p28, RT activity, and the 3.4-kb RNA transcript. RT-PCR and in vitro translation indicate that some of the classic HTLV-I particles package 3.4-kb RNA as well as full-length 8.5-kb RNA. In addition to matrix features, the p28 protein has a motif resembling a zinc finger at the C-terminal, pX0 region, which may play a role in the assembly of the defective light virions. (C) 1999 Academic Press.

关键词：细胞系转化离心法梯密度化学分馏法基因产物 gag/分离和提纯人类嗜T淋巴细胞病毒1型/遗传学人类嗜T淋巴细胞病毒1型/分离和提纯人类嗜T淋巴细胞病毒1型/代谢 RNA指导的DNA聚合酶/代谢逆转录病毒科蛋白质类致癌/分离和提纯病毒粒子/分离和提纯人类

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Generation of transmissible hepatitis C virions from a molecular clone in chimpanzees

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VIROLOGY 1999年第1期256卷 36-44页

作者： Hong, Z Beaudet-Miller, M Lanford, RE Guerra, B Wright-Minogue, J Skelton, A Baroudy, BM Reyes, GR Lau, JYN Schering Plough Corp Inst Res Antiviral Therapy Kenilworth NJ 07033 USA SW Fdn Biomed Res Dept Virol & Immunol San Antonio TX 78227 USA

Multiple alignments of hepatitis C virus (HCV) polyproteins from six different genotypes identified a total of 22 nonconsensus mutations in a clone derived from the Hutchinson (H77) isolate. These mutations, collectively, may have contributed to the failure in generating a "functionally correct" or "infectious" clone in earlier attempts. A consensus clone was constructed after systematic repair of these mutations, which yielded infectious virions in a chimpanzee after direct intrahepatic inoculation of in vitro transcribed RNAs. This RNA-infected chimpanzee has developed hepatitis and remained HCV positive for more than 11 months. To further verify this RNA-derived infectivity, a second naive chimpanzee was injected intravenously with serum collected from the first chimpanzee. Infectivity analysis of the second chimpanzee demonstrated that the HCV infection was successfully transmitted, which validated unequivocally the infectivity of our repaired molecular clone. Amino acid sequence comparisons revealed that our repaired infectious clone had 4 mismatches with the isogenic clone reported by Kolykhalov et al. (1997 Science 277, 570-574) and 8 mismatches with that reported by Yanagi et ai (1997, Proc. Natl. Acad. Sci. USA 94, 8738-8743). At the RNA level, more mismatches (43 and 67, respectively) were identified;most of them were synonymous substitutions. Further comparisons with 16 isolates from different genotypes demonstrated that our repaired clone shares greater consensus than the reported isogenic clones. This approach of generating infectious HCV RNA validates the importance of amino acid sequence consensus in relation to the biology of HCV. (C) 1999 Academic Press.

关键词：氨基酸序列氨基酸取代克隆分子 DNA修复肝炎病毒属/遗传学肝炎病毒属/分离和提纯肝炎丙型/病理学肝/病理学肝/病毒学分子序列数据突变黑猩猩属/病毒学 RNA 病毒/遗传学逆转录聚合酶链反应序列比对转录遗传转染病毒核心蛋白质类/化学病毒核心蛋白质类/遗传学病毒非结构蛋白质类/化学病毒非结构蛋白质类/遗传学病毒粒子/遗传学病毒粒子/分离和提纯动物

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Infectious hypodermal and hematopoietic necrosis virus of shrimp is related to mosquito Brevidensoviruses

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VIROLOGY 2000年第1期277卷 167-177页

作者： Shike, H Dhar, AK Burns, JC Shimizu, C Jousset, FX Klimpel, KR Bergoin, M Univ Calif San Diego Sch Med Dept Pediat La Jolla CA 92093 USA Super Shrimp Grp Inc National City CA 91950 USA Univ Montpellier 2 Mol Virol Unit UMR 5087 F-34095 Montpellier France

We purified and sequenced infectious hypodermal and hematopoietic necrosis virus (IHHNV), a small DNA virus of shrimp, from wild Penaeus stylirostris. The virion has a buoyant density of 1.45 as determined by cesium chloride gradient. Analysis of 3873 nucleotides of the viral genome revealed three large open reading frames (ORFs) and parts of the noncoding termini of the viral genome. The left, mid, and right ORFs on the complementary (plus) strand have potential coding capacities of 666 amino acids (aa) (75.77 kDa), 363 aa (42.11 kDa), and 329 aa (37.48 kDa), respectively. The overall genomic organization is similar to that of the mosquito brevidensoviruses. The left ORF most likely encodes the major nonstructural (NS) protein (NS-1) since it contains conserved replication initiator motifs and NTP-binding and helicase domains similar to those in NS-1 from all other parvoviruses. The IHHNV putative NS-1 shares the highest aa sequence homology with the NS-1 of mosquito brevidensoviruses, Aedes densovirus and Aedes albopictus parvovirus. A search for putative splicing sites revealed that the N-terminal region of NS-1 is very likely located in a small ORF upstream of the left ORF. The right ORF is presumed to encode structural polypeptides (VPs), as in other parvoviruses. Two putative promoters, located upstream of the left and right ORFs, are presumed to regulate expression of NS and VP genes, respectively. Thus, IHHNV is closely related to densoviruses of the genus Brevidensovirus in the family Parvoviridae, and we therefore propose to rename this virus Penaeus stylirostris densovirus(PstDNV). (C) 2000 Academic Press.

关键词：氨基酸序列碱基序列蚊科/病毒学 DNA病毒/分类 DNA病毒/遗传学 DNA病毒/生理学十足目(甲壳纲)/病毒学基因组病毒分子序列数据开放读码框架种系发生聚合酶链反应序列比对序列同源性氨基酸病毒非结构蛋白质类/化学病毒非结构蛋白质类/遗传学病毒粒子/分类病毒粒子/遗传学病毒粒子/分离和提纯病毒复制动物

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Purification, crystallization and preliminary X-ray analysis of immunogenic viruslike particles formed by infectious bursal disease virus (IBDV) structural protein VP2

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ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY 2003年第7期59卷 1234-1237页

作者： Lee, CC Ko, TP Lee, MS Chou, CC Lai, SY Wang, AHJ Wang, MY Natl Chung Hsing Univ Grad Inst Biotechnol Taichung 40227 Taiwan Acad Sinica Inst Biol Chem Taipei 11529 Taiwan Chungtai Inst Hlth Sci & Technol Dept Food Sci Taichung 40605 Taiwan

Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry VP2 protein, which consists of 452 amino-acid residues, is the primary immunogen of IBDV and contains the epitopes responsible for eliciting neutralizing antibodies. When the chimeric VP2 protein (rVP2H) of a local IBDV strain P3009 was expressed alone using the baculovirus system, virus-like particles of approximately 23 nm in diameter formed spontaneously. Highly pure rVP2H particles, obtained using ammonium sulfate precipitation, immobilized metal-ion affinity chromatography and gel-filtration chromatography, were successfully crystallized using the vapour-diffusion method. These crystals, with a maximum dimension of 0.4 mm, diffracted X-rays to 4.5 Angstrom resolution, but data were only collected to 6 Angstrom. Preliminary analysis of the diffraction data showed that the rVP2H crystals belong to the cubic space group P2(1)3, with unit-cell parameter 323.1 Angstrom. The icosahedral symmetry of the particles is clearly seen in the self-rotation function maps, with dyads and triads coincident with the crystallographic axes. Each asymmetric unit contains 1/3 of the particle, or 20 rVP2H subunits, and there are four particles in a unit cell, probably in a tetrahedral arrangement.

关键词：杆状病毒科/遗传学克隆分子结晶结晶学 X线传染性腔上囊病病毒/化学重组蛋白质类病毒结构蛋白质类/化学病毒结构蛋白质类/免疫学病毒粒子/化学病毒粒子/免疫学病毒粒子/分离和提纯

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PAPOVAVIRAL SIALOADENITIS IN ATHYMIC NUDE RATS

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LABORATORY ANIMALS 1984年第1期18卷 84-89页

作者： WARD, JM LOCK, A COLLINS, MJ GONDA, MA REYNOLDS, CW NCI BIOL RESPONSE MODIFIERS PROGRAM FREDERICK MD 21701 USA NIH DIV RES SERV VET RESOURCES BRANCH COMPARAT PATHOL SECT BETHESDA MD 20205 USA NCI FREDERICK CANC RES FACIL PROGRAM RESOURCES INC ANIM HLTH DIAGNOST LAB FREDERICK MD 21701 USA NCI FREDERICK CANC RES FACIL PROGRAM RESOURCES INC ELECTRON MICROSCOPY LAB FREDERICK MD 21701 USA

A wasting disease was found in 32 athymic nude rats. The rats had parotid sialoadenitis with intranuclear inclusion bodies in ductal and acinar epithelial cells. Other common lesions included bronchitis, bronchiolitis and secondary bacterial pneumonia. Less commonly, rhinitis and Harderian adenitis were seen. Intranuclear inclusions were also seen in bronchial epithelium of 1 rat, Harderian gland acini of 1 rat and laryngeal glands of 2 rats. Viral particles, averaging 45 nm in diameter, sometimes in crystalline arrays, were found in the nucleus of parotid epithelial cells. By the use of the avidin-biotin-peroxidase complex (ABC) immunoperoxidase technique, antibodies to disrupted SV40 virus (the group specific antigen of the polyoma virus (miopapova virus) genus of the papova virus family) reacted with intranuclear inclusions and cytoplasm of parotid epithelium and inclusions in lung and Harderian gland. The viral antigen did not cross-react with antibodies to mouse polyoma, mouse K or disrupted bovine papillomaviruses.

关键词：包涵体病毒/超微结构包涵体病毒/病毒学腮腺/病毒学多瘤病毒属/分类多瘤病毒属/分离和提纯多瘤病毒属/超微结构多瘤病毒感染/病理学多瘤病毒感染/兽医学多瘤病毒感染/病毒学大鼠裸啮齿动物疾病/病理学啮齿动物疾病/病毒学涎腺炎/病理学涎腺炎/兽医学涎腺炎/病毒学病毒粒子/分类病毒粒子/分离和提纯病毒粒子/超微结构动物女(雌)性男(雄)性大鼠

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