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Expression of K13/v-FLIP gene of human herpesvirus 8 and apoptosis in Kaposi's sarcoma spindle cells

JOURNAL OF THE NATIONAL CANCER INSTITUTE 1999年第20期91卷 1725-1733页

作者： Stürzl, M Hohenadl, C Zietz, C Castanos-Velez, E Wunderlich, A Ascherl, G Biberfeld, P Monini, P Browning, PJ Ensoli, B GSF Forschungszentrum Umwelt & Gesundheit GMBH Inst Mol Virol D-85764 Neuherberg Germany Max Planck Inst Biochem Dept Virus Res D-82152 Martinsried Germany Bavarian Nord Res Inst AS Martinsried Germany Univ Munich Inst Pathol D-8000 Munich Germany Karolinska Inst Immunopathol Lab Stockholm Sweden Tech Univ Munich Inst Virol D-8000 Munich Germany Ist Super Sanita Virol Lab I-00161 Rome Italy Vanderbilt Univ Nashville TN USA

Background: Human herpesvirus 8 (HHV8) infection is associated with all forms of Kaposi's sarcoma (KS), The HHV8 genome locus ORFK13-72-73 (ORF = open reading frame) encodes proteins that may be important in HHV8-mediated pathogenesis, i.e., the latency-associated nuclear antigen (encoded by ORF73), viral-cyc-D (v-cyc-D), a viral homologue of cellular cyclin D (encoded by ORF72), and viral-FLIP (v-FLIP), a homologue of the cellular FLICE (Fas-associated death domain-like interleukin 1 beta-converting enzyme) inhibitory protein (encoded by ORFK13;is an inhibitor of apoptosis [programmed cell death]), Through differential splicing events, this locus expresses individual RNA transcripts that encode all three proteins (tricistronic transcripts) or just two of them (v-FLIP and v-cyc-D;bicistronic transcripts). We examined expression of these transcripts in KS tissues. Methods: We collected tissues from patients with KS of different stages. By use of an optimized in situ hybridization procedure, we examined different ORFK13-72-73 locus transcripts in HHV8-infected cells in skin lesions and in one adjacent lymph node. Apoptosis in KS lesions was determined by use of an in situ assay. Results and Conclusions: Our results indicate the following: 1) Transcripts from the ORFK13-72-73 locus appear to be spliced differentially in latently infected KS cells in skin lesions and in HHV8-infected cells in lymph nodes;specifically, ORFK13-ORF72 bicistronic transcripts were expressed abundantly in KS cells, whereas ORFK13-ORF72-ORF73 tricistronic transcripts were detected only in lymph node cells. 2) Sequences encoding the antiapoptotic protein v-FLIP are expressed at very low levels in early KS lesions, but expression increases dramatically in late-stage lesions. 3) The increase in expression of v-FLIP-encoding transcripts is associated with a reduction in apoptosis in KS lesions. Implications: These data suggest that functional v-FLIP is produced in vivo and that antiapoptotic mecha

关键词：抗原病毒/分析抗原病毒/遗传学细胞凋亡 CASP8和FADD样细胞凋亡调节蛋白质载体蛋白质类/分析载体蛋白质类/遗传学减量调节基因表达基因病毒疱疹病毒8型人/遗传学免疫组织化学原位杂交原位缺口末端标记细胞内信号肽和蛋白质类淋巴结/代谢淋巴结/病毒学肿瘤分期核蛋白质类/分析核蛋白质类/遗传学开放读码框架 RNA探针 RNA 信使/分析 RNA 肿瘤/分析 RNA 病毒/分析肉瘤卡波西/遗传学肉瘤卡波西/病理学肉瘤卡波西/病毒学转录遗传增量调节病毒蛋白质类/遗传学人类

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Comparison of human sera reactivities in immunoblots with recombinant human herpesvirus (HHV)-8 proteins associated with the latent (ORF73) and lytic (ORFs 65, K8.1A, and K8.1B) replicative cycles and in immunofluorescence assays with HHV-8-infected BCBL-1 cells

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VIROLOGY 1999年第2期256卷 381-392页

作者： Zhu, LJ Wang, R Sweat, A Goldstein, E Horvat, R Chandran, B Univ Kansas Med Ctr Dept Microbiol Mol Genet & Immunol Kansas City KS 66160 USA Univ Kansas Med Ctr Dept Med Div Infect Dis Kansas City KS 66160 USA Univ Kansas Med Ctr Dept Pathol Div Infect Dis Kansas City KS 66160 USA Univ Kansas Med Ctr Clin Labs Kansas City KS 66160 USA

The development of reliable, sensitive, and specific serological methods for the detection of human herpesvirus-8 (HHV-8) antibodies is critical for a thorough understanding of HHV-8 prevalence and pathogenesis. To evaluate the potential usefulness of HHV-8 proteins in measuring the responses against both latent and lytic antigens, we selected 1 latent [open reading frame (ORF) 73] antigen and 3 HHV-8 lytic antigens (ORFs 65, K8.1A, and K8.1B) previously identified as immunogenic [Virology (1998) 243, 208-217]. Full-length genomic ORF 73 and full-length ORFs 65, K8.1A, and K8.1B from the cDNA clones were cloned, expressed in bacterial and baculovirus-insect cell expression systems, and purified as GST fusion proteins. These recombinant proteins were used in Western blot reactions to test sera from 104 human immunodeficiency virus (HIV)(+)/Kaposi's sarcoma (KS)(+) homosexual men, 77 HIV+/KS- homosexual men, and 84 age-matched HIV-/KS- men. These Sera were also tested in immunofluorescence assays (IFAs) with uninduced and 12-O-tetradecanoylphorbol-13-acetate-induced B cell lymphoma-1 cells to detect antibodies against latency-associated nuclear antigens (LANA) and antibodies against lytic antigens (cytoplasmic fluorescence). These sera exhibited differential reactivities reflecting different titers of antibodies against HHV-8 proteins, and Variable reactivities were seen more commonly with the sera from HIV-/KS- adult men. In the Western blot assay, 89% (93 of 104) of HIV+/KS (+) sera, 60% (46 of 77) of HIV+/KS- sera, and 7% (6 of 84) HIV+/KS- sera were reactive with both latent and lytic recombinant antigens. Western blot reactions with ORF 73 protein were more sensitive than LANA-IFA results. The lytic IFA and lytic Western blot (ORFs 65 and K8.1A) assays were more sensitive than the ORF 73 Western blots and LANA-IFA. With an exception of 2 sera from the HIV-/KS- group, all sera positive for lytic IFA antibodies and ORF 65 and K8.1A antibodies were also positive for

关键词：抗体病毒/血液抗体病毒/免疫学抗原病毒/遗传学抗原病毒/免疫学印迹法蛋白质细胞系荧光抗体技术间接基因表达谷胱甘肽转移酶/遗传学谷胱甘肽转移酶/免疫学糖蛋白类/遗传学糖蛋白类/免疫学 HIV感染/免疫学疱疹病毒8型人/遗传学疱疹病毒8型人/免疫学免疫印迹法核蛋白质类/遗传学核蛋白质类/免疫学开放读码框架磷蛋白类重组融合蛋白质类/遗传学重组融合蛋白质类/免疫学重组遗传肉瘤卡波西/血液肉瘤卡波西/免疫学肉瘤卡波西/病毒学病毒蛋白质类/遗传学病毒蛋白质类/免疫学病毒潜伏期病毒复制青少年成年人人类男(雄)性中年人

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Molecular evidence of organ-related transmission of Kaposi sarcoma-associated herpesvirus or human herpesvirus-8 in transplant patients

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BLOOD 2000年第9期96卷 3279-3281页

作者： Luppi, M Barozzi, P Santagostino, G Trovato, R Schulz, TF Marasca, R Bottalico, D Bignardi, L Torelli, G Univ Modena Dept Med Sci Sect Hematol I-41100 Modena Italy Univ Parma Div Nephrol I-43100 Parma Italy Univ Liverpool Dept Med Microbiol & Genitourinary Med Liverpool L69 3BX Merseyside England

In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus or human herpesvirus-8 (HHV-8) infection is associated with the development of KS, primary effusion lymphoma and Castleman disease. Whether HHV-8 is either reactivated in the recipient or transmitted by the donor has been investigated so far only by serologic studies. Thus, we addressed the issue of HHV-8 transmission in the transplantation setting by molecular methods. We exploited the high level variability of the orf-K1 gene and the polymorphism of the orf-73 gene of the HHV-8 genome to assess the genetic relatedness of the HHV-8 strains identified in the posttransplant KS lesions that developed, simultaneously, 20 months after transplantation, in 2 recipients of twin kidneys from the same cadaver donor. The 100% identity of nucleotide sequence of the most variable viral region and the presence of the same, single orf-73 type in both patients provides strong molecular evidence of organ-related transmission of HHV-8 in the setting of transplantation. (C) 2000 by The American Society of Hematology.

关键词：碱基序列基因型疱疹病毒8型人/遗传学疱疹病毒8型人/分离和提纯疱疹病毒8型人/生理学肾移植分子序列数据手术后并发症肉瘤卡波西/病毒学序列比对病毒激活女(雌)性人类中年人

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KS enters Y2K still riddled with many questions

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JOURNAL OF THE NATIONAL CANCER INSTITUTE 1999年第19期91卷 1612-1614页

作者： Miller, M

Discusses the mysteries surrounding Kaposi's sarcoma (KS), an AIDS-related malignancies. How Robert Gallo, director of the Institute of Human Virology in Baltimore, Maryland, described KS; Puzzling aspects of KS; ... 详细信息

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Differential viral protein expression in Kaposi's sarcoma-associated herpesvirus-infected diseases - Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease

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AMERICAN JOURNAL OF PATHOLOGY 2000年第3期156卷 743-749页

作者： Parravicini, C Chandran, B Corbellino, M Berti, E Paulli, M Moore, PS Chang, Y Columbia Univ Coll Phys & Surg Dept Pathol New York NY 10032 USA Lugio Sacco Hosp Dept Pathol Milan Italy Univ Kansas Med Ctr Dept Microbiol Mol Genet & Immunol Kansas City KS 66103 USA Univ Milan Luigi Sacco Hosp Inst Infect Dis & Trop Med Milan Italy Univ Pavia IRCCS Osped San Matteo Dept Pathol I-27100 Pavia Italy Columbia Univ Div Epidemiol New York NY USA

Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is Linked to KS, primary effusion lymphomas (PEL), and a subset of multicentric Castleman's disease (MCD), Transcript mapping studies using PEL cell lines have allowed preliminary classification of viral gene expression into constitutive (class I) and inducible (class II/III) categories. To determine whether viral gene expression differs in vivo, we examined tissue sections of KSHV-infected disorders, using specific antibodies against proteins that are representative of the different expression classes of KSHV genes. ORF73/LANA appears to be a surrogate marker for KSHV infection because it is constitutively expressed in vitro and in vivo in all KSHV-infected tells. Expression of vIRF1, vIL6, and PF-8 proteins in the infected B cells of MCD lymph nodes reproduces the expression pattern observed in TPA-stimulated KSHV-infected B-cell lines. In contrast, the protein. expression of the inducible viral genes that me tested in KS and PEL biopsies is restricted to PF-8 and vIL6, respectively. The tightly restricted expression of KSHV proteins in vivo differs from the dysregulated expression of inducible KSHV genes in vitro and suggests that viral gene expression in KSHV-infected cell lines does not accurately reflect what occurs in diseased tissues. These differences may be related to either cell-specific or immune restriction of viral replication.

关键词：荧光抗体技术间接基因表达基因病毒巨淋巴结增生/代谢巨淋巴结增生/病理学巨淋巴结增生/病毒学疱疹病毒8型人/遗传学疱疹病毒8型人/分离和提纯淋巴结/病理学淋巴结/病毒学淋巴瘤 B细胞/代谢淋巴瘤 B细胞/病理学淋巴瘤 B细胞/病毒学肉瘤卡波西/代谢肉瘤卡波西/病理学肉瘤卡波西/病毒学皮肤肿瘤/代谢皮肤肿瘤/病理学皮肤肿瘤/病毒学肿瘤细胞培养的病毒蛋白质类/代谢人类

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Human herpesvirus 8 seropositivity and risk of Kaposi's sarcoma and other acquired immunodeficiency syndrome-related diseases

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JOURNAL OF THE NATIONAL CANCER INSTITUTE 1999年第17期91卷 1468-1474页

作者： Rezza, G Andreoni, M Dorrucci, M Pezzotti, P Monini, P Zerboni, R Salassa, B Colangeli, V Sarmati, L Nicastri, E Barbanera, M Pristerà, R Aiuti, F Ortona, L Ensoli, B Ist Super Sanita Ctr Operat AIDS Epidemiol & Biostat Lab I-00161 Rome Italy Ist Super Sanita Virol Lab I-00161 Rome Italy Univ Roma Tor Vergata Dept Infect Dis Rome Italy Univ Milan Sexually Transmitted Dis Clin I-20122 Milan Italy Osped Amedeo di Savoia Div Infect Dis Turin Italy Univ Bologna Infect Dis Clin I-40126 Bologna Italy Osped Riuniti Bergamo Div Infect Dis Livorno Italy Osped Reg Div Infect Dis Bolzano Italy Univ Roma La Sapienza Rome Italy Univ Cattolica Sacro Cuore I-00168 Rome Italy

Background: The incidence of Kaposi's sarcoma (KS) is increased severalfold in individuals infected with human immunodeficiency virus-1 (HIV). Human herpesvirus 8 (HHVS) has also been implicated in KS. We investigated several factors that may determine the onset of KS, particularly HHVS infection in individuals after becoming seropositive for HIV. Methods: We studied 366 individuals belonging to different HIV-exposure categories (i.e., homosexual activity, intravenous drug use, and heterosexual contact) for whom a negative HIV serologic test and then a positive HIV serologic test were available within a 2-year period. HHV8 antibody testing was performed by use of an immunofluorescence assay on the first serum sample available after the first positive HIV test. Actuarial rates of progression of KS and of other acquired immunodeficiency syndrome (AIDS)-defining diseases were estimated by use of time-to-event statistical methods. All statistical tests were two-sided. Results: Twenty-one of the 366 study participants developed AIDS-related KS, and 83 developed AIDS without KS. One hundred forty (38.3%) participants had detectable anti-HHV8 antibodies. The actuarial progression rate to KS among persons co-infected with HIV/HHV8 was nearly 30% by 10 years after HIV seroconversion. Increasing HHV8 antibody titers increased the risk of developing KS (for seronegative versus highest titer [1 :125 serum dilution], adjusted relative hazard [RH] = 51.82;95% confidence interval [CI] = 6.08-441.33) but not of other AIDS-defining diseases (adjusted RH = 1.14;95% CI = 0.72-1.80). HHV8-seropositive homosexual men compared with HHV8-seropositive participants from other HIV-exposure categories showed an increased risk of KS that approached statistical significance (adjusted RH = 6.93;95% CI = 0.88-54.84). Conclusions: Approximately one third of individuals coinfected with HIV/HHV8 developed KS within 10 years after HIV seroconversion. Progression to KS increased with time after HIV se

关键词：艾滋病相关机会致病菌感染/病毒学保险统计分析抗体病毒/血液 CD4淋巴细胞计数疾病恶化 HIV感染/并发症疱疹病毒科感染/并发症疱疹病毒4型人/免疫学疱疹病毒8型人/免疫学意大利危险肉瘤卡波西/病毒学青少年成年人老年人女(雌)性人类男(雄)性中年人

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Human herpesvirus 8 and cancer

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JOURNAL OF THE NATIONAL CANCER INSTITUTE 1999年第17期91卷 1440-1441页

作者： Beral, V Newton, R Sitas, F Univ Oxford Imperial Canc Res Fund Canc Epidemiol Unit Oxford England S African Inst Med Res Natl Canc Registry Canc Epidemiol Unit ZA-2000 Johannesburg South Africa

Beral et al comment on some research articles on a new herpesvirus in Kaposi's sarcoma tumor known as human herpesvirus 8 (HHV8).

关键词：抗体病毒/血液 HIV感染/并发症 HIV感染/免疫学 HIV感染/病毒学疱疹病毒8型人/免疫学发病率意大利危险因素肉瘤卡波西/并发症肉瘤卡波西/免疫学肉瘤卡波西/病毒学人类

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Perspective on Kaposi's sarcoma: Facts, concepts, and conjectures

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JOURNAL OF THE NATIONAL CANCER INSTITUTE 1999年第17期91卷 1453-1458页

作者： Reitz, MS Nerurkar, LS Gallo, RC Univ Maryland Inst Human Virol Baltimore MD 21201 USA NCI Int Canc Informat Ctr Bethesda MD 20892 USA

Kaposi's sarcoma (KS) is the most common tumor associated with HIV-1 infection. Information on KS, including the role of the HIV-1 Tat protein and KS cell types, is provided.

关键词：基因产物 tat/代谢生长物质/生物合成 HIV感染/并发症疱疹病毒8型人/代谢肉瘤卡波西/代谢肉瘤卡波西/病理学肉瘤卡波西/病毒学人类

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Human herpesvirus 8 infections in the Amsterdam Cohort Studies (1984-1997): Analysis of seroconversions to ORF65 and ORF73

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第9期97卷 4838-4843页

作者： Goudsmit, J Renwick, N Dukers, NHTM Coutinho, RA Heisterkamp, S Bakker, M Schulz, TF Cornelissen, M Weverling, GJ Univ Amsterdam Acad Med Ctr Dept Human Retrovirol NL-1105 AZ Amsterdam Netherlands Municipal Hlth Serv Dept Publ Hlth & Environm Amsterdam Netherlands Univ Amsterdam Acad Med Ctr Dept Clin Epidemiol & Biostat NL-1105 AZ Amsterdam Netherlands Univ Liverpool Dept Med Microbiol & Genitourinary Med Liverpool L69 3BX Merseyside England

We have shown previously that human herpesvirus 8 (HHV8) seroconversion for antibodies to the latency-associated nuclear antigen encoded by ORF73 and/or the lytic capsid antigen (vp19) encoded by ORF65 is associated with orogenital contact and is strongly linked to the development of Kaposi's sarcoma among HIV-infected individuals in the Amsterdam Cohort Studies. Here, we investigate the relationship between seroconversion to these antigens and primary HHV8 infection. Between 1984 and 1997, 215 HHV8 seroconversions to ORF73 (106 cases or 49%) and/or to ORF65 (159 cases or 74%) were recorded in the cohort of homosexual men. The HHV8 seroconversion rate among HIV-infected homosexual men (6.2 per 100 person years) was consistently higher than among HIV-uninfected men (2.6 per 100 person years). In HIV-infected but not in uninfected individuals, seroconversion to ORF73/latency-associated nuclear antigen precedes that to ORF65/vp19. Antibody levels to both ORF65- and ORF73-encoded antigens were higher in HIV-infected than in HIV-uninfected men, and among HIV-seropositives, antibody levels to ORF65/vp19 rise even higher with declining CD4 cell counts and peak with Kaposi's sarcoma development, suggesting continuing and increasing viral replication. In 10.3% of HHV8 seroconversions, transient serum viremia could be demonstrated before or at seroconversion. Together with the previously reported link between unprotected orogenital sex and HHV8 seroconversion, our observations suggest that HHV8 seroconversions result from primary infections.

关键词：抗体病毒/血液抗体生成抗原病毒/免疫学队列研究 HIV感染/并发症 HIV感染/流行病学疱疹病毒科感染/流行病学疱疹病毒科感染/免疫学疱疹病毒科感染/病毒学疱疹病毒8型人/遗传学疱疹病毒8型人/分离和提纯同性恋男性荷兰/流行病学核蛋白质类/免疫学肉瘤卡波西/流行病学肉瘤卡波西/免疫学肉瘤卡波西/病毒学病毒蛋白质类/免疫学人类男(雄)性

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Pretransplantation human herpes virus 8 seropositivity as a risk factor for Kaposi's sarcoma in kidney transplant recipients

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TRANSPLANTATION PROCEEDINGS 2000年第3期32卷 526-527页

作者： Cattani, P Nanni, G Graffeo, R Capuano, M Cerimele, F La Parola, IL Diociaiuti, A Pozzetto, U Cerimele, D Fadda, G Castagneto, M Univ Cattolica Sacro Cuore Inst Microbiol Transplant Unit Inst Dermatol Rome Italy

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