scholarly journals Characterization of Entry Mechanisms of Human Herpesvirus 8 by Using an Rta-Dependent Reporter Cell Line

2003 ◽  
Vol 77 (14) ◽  
pp. 8147-8152 ◽  
Author(s):  
Naoki Inoue ◽  
Jörn Winter ◽  
Renu B. Lal ◽  
Margaret K. Offermann ◽  
Shin Koyano
Keyword(s):  
Cell Line ◽  
Primary Effusion Lymphoma ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Dependent Manner ◽  
Reporter Cell Line ◽  
Reporter Cell ◽  
Herpesvirus 8 ◽  
Intracellular Expression ◽  
Infected Cells

ABSTRACT To analyze the mechanisms of entry of human herpesvirus 8 (HHV-8), we established a reporter cell line T1H6 that contains the lacZ gene under the control of the polyadenylated nuclear RNA promoter, known to be strongly activated by a viral transactivator, Rta. We found that infection with cell-free virus, as well as cocultivation with HHV-8-positive primary effusion lymphoma cell lines, activated the lacZ gene of T1H6 in a sensitive and dose-dependent manner. Addition of Polybrene and centrifugation enhanced, but polysulfonate compounds inhibited, the HHV-8 infectivity. RGD-motif-containing polypeptides and integrins did not decrease the infectivity, suggesting the presence of an additional cellular receptor other than the reported one. The entry was dependent on pH acidification but not on the clathrin pathway. Although conditioned media obtained from human immunodeficiency virus (HIV)-infected cells did not have any effect on the early steps of HHV-8 infection, intracellular expression of a proviral HIV type 1, but not of Tat alone, increased the HHV-8-dependent reporter activation slightly, suggesting a potential of HIV-mediated enhancement of an early step of HHV-8 infection.

scholarly journals Characterization of Entry Mechanisms of Human Herpesvirus 8 by Using an Rta-Dependent Reporter Cell Line

2003 ◽  
Vol 77 (18) ◽  
pp. 10177-10177 ◽  
Author(s):  
Naoki Inoue ◽  
Jörn Winter ◽  
Renu B. Lal ◽  
Margaret K. Offermann ◽  
Shin Koyano
Keyword(s):  
Cell Line ◽  
Human Herpesvirus ◽  
Human Herpesvirus 8 ◽  
Reporter Cell Line ◽  
Reporter Cell ◽  
Herpesvirus 8

scholarly journals A Systems Biology Approach To Identify the Combination Effects of Human Herpesvirus 8 Genes on NF-κB Activation

2009 ◽  
Vol 83 (6) ◽  
pp. 2563-2574 ◽  
Author(s):  
Andreas Konrad ◽  
Effi Wies ◽  
Mathias Thurau ◽  
Gaby Marquardt ◽  
Elisabeth Naschberger ◽  
...  
Keyword(s):  
Systems Biology ◽  
Primary Effusion Lymphoma ◽  
Human Herpesvirus ◽  
Etiologic Agent ◽  
Lytic Replication ◽  
Fine Tuning ◽  
Human Herpesvirus 8 ◽  
Systematic Analysis ◽  
Herpesvirus 8 ◽  
Infected Cells

ABSTRACT Human herpesvirus 8 (HHV-8) is the etiologic agent of Kaposi's sarcoma and primary effusion lymphoma. Activation of the cellular transcription factor nuclear factor-kappa B (NF-κB) is essential for latent persistence of HHV-8, survival of HHV-8-infected cells, and disease progression. We used reverse-transfected cell microarrays (RTCM) as an unbiased systems biology approach to systematically analyze the effects of HHV-8 genes on the NF-κB signaling pathway. All HHV-8 genes individually (n = 86) and, additionally, all K and latent genes in pairwise combinations (n = 231) were investigated. Statistical analyses of more than 14,000 transfections identified ORF75 as a novel and confirmed K13 as a known HHV-8 activator of NF-κB. K13 and ORF75 showed cooperative NF-κB activation. Small interfering RNA-mediated knockdown of ORF75 expression demonstrated that this gene contributes significantly to NF-κB activation in HHV-8-infected cells. Furthermore, our approach confirmed K10.5 as an NF-κB inhibitor and newly identified K1 as an inhibitor of both K13- and ORF75-mediated NF-κB activation. All results obtained with RTCM were confirmed with classical transfection experiments. Our work describes the first successful application of RTCM for the systematic analysis of pathofunctions of genes of an infectious agent. With this approach, ORF75 and K1 were identified as novel HHV-8 regulatory molecules on the NF-κB signal transduction pathway. The genes identified may be involved in fine-tuning of the balance between latency and lytic replication, since this depends critically on the state of NF-κB activity.

scholarly journals Kaposi sarcoma–associated herpesvirus/human herpesvirus 8–associated lymphoproliferative disorders

Blood ◽  
2019 ◽  
Vol 133 (11) ◽  
pp. 1186-1190 ◽  
Author(s):  
Eric Oksenhendler ◽  
David Boutboul ◽  
Lionel Galicier
Keyword(s):  
B Cells ◽  
Primary Effusion Lymphoma ◽  
Human Herpesvirus ◽  
Kaposi Sarcoma ◽  
Castleman Disease ◽  
Lymphoproliferative Disorders ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Multicentric Castleman Disease ◽  
Infected Cells

Abstract Kaposi sarcoma–associated herpesvirus/human herpesvirus 8 is associated with multicentric Castleman disease (MCD) and primary effusion lymphoma (PEL). In MCD, infected B cells, although polyclonal, express a monotypic immunoglobulin Mλ phenotype, probably through editing toward λ light chain in mature B cells. They are considered to originate from pre–germinal center (GC) naive B cells. Both viral and human interleukin-6 contribute to the plasmacytic differentiation of these cells, and viral replication can be observed in some infected cells. PEL cells are clonal B cells considered as GC/post-GC B cells. One can also hypothesize that they originate from the same infected naive B cells and that additional factors could be responsible for their peculiar phenotype.

scholarly journals Generation of a Reporter Cell Line for Detection of Infectious Varicella-Zoster Virus and Its Application to Antiviral Studies

2006 ◽  
Vol 50 (9) ◽  
pp. 3142-3145 ◽  
Author(s):  
Guan-Qing Wang ◽  
Tatsuo Suzutani ◽  
Yumiko Yamamoto ◽  
Yoshiko Fukui ◽  
Naoki Nozawa ◽  
...  
Keyword(s):  
Varicella Zoster Virus ◽  
Cell Line ◽  
Dependent Manner ◽  
Reporter Cell Line ◽  
Reporter Cell ◽  
Varicella Zoster ◽  
Infected Cells ◽  
Dose Dependent

ABSTRACT To simplify the titration of infectious varicella-zoster virus (VZV), we generated a reporter cell line that produced luciferase in a dose-dependent manner upon infection with cell-free VZV. A few VZV-infected cells were detectable by coculturing with the cell line. We demonstrated the usefulness of the cell line for antiviral studies.

Human Herpesvirus 8 K1 Blocks Fas in Cells and Mice by Binding and Blocking Fas.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 515-515
Author(s):  
Hoyoung Maeng ◽  
Suizhao Wang ◽  
Zuzuna Berkova ◽  
Shu Wang ◽  
Jaehoon Jung ◽  
...  
Keyword(s):  
Kaposi's Sarcoma ◽  
Primary Effusion Lymphoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Dependent Manner ◽  
Intact Cells ◽  
Herpesvirus 8 ◽  
In Cells

Abstract Background: Kaposi’s sarcoma and primary effusion lymphoma are linked to human herpesvirus 8 (HHV-8 or Kaposi’s sarcoma-associated virus) infection. Key to the pathogenesis of these cancers is the HHV-8 transmembrane oncoprotein K1. This viral protein, which immortalizes cells, produces lymphoproliferation and lymphomas through unknown mechanisms when expressed in mice. Results: We show here that K1 blocks Fas (CD95)-mediated apoptosis by binding the ectodomain of K1 to Fas in hematopoietic cancer cells (BJAB, U937 and THP-1). K1 bound to Fas in K1-transfected BJAB cells. To assess whether K1 and Fas formed complexes in cells, we chemically crosslinked proteins in transfected cells using 3,3′-dithiobissulfosuccinimidyl propionate in the presence of N-ethyl maleimide. Stabled K1-Fas complexes were present in intact cells despite blocking of free reactive cysteines. The selective ability of K1 to antagonize Fas-mediated apoptosis was strictly dependent on having its immunoreceptor tyrosine-based activation motif (ITAM). Using activating anti-Fas (CH-11) and nonactivating anti-Fas (B10) antibodies in a serial manner, we showed that K1 associated with Fas that was not activated, strongly suggesting that K1 prevents Fas from being activated. Mice transfected with K1 were protected from the apoptosis inducing effects of agonistic Jo2 anti-Fas antibody and tissues from these mice contained K1-Fas complexes. K1-transfected compared to vector-transfected mice had fewer terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive cells (13 ± 3% vs. 60 ± 7%, P=0.008), fewer activated caspase 3 positive cells (24 ± 5% vs. 88 ± 6%, P=0.006). Tissues had less hemorrhaging, and necrosis. Conclusion: These data support a mechanism of K1 direct binding to and blockage by K1 to Fas in suppression of apoptosis in an ITAM-dependent manner, which may be a key step in lymphoma development.

Association of Primary Pleural Effusion Lymphoma of T-Cell Origin and Human Herpesvirus 8 in a Human Immunodeficiency Virus–Seronegative Man

2001 ◽  
Vol 125 (9) ◽  
pp. 1246-1248 ◽  
Author(s):  
Emmanuèle Lechapt-Zalcman ◽  
Dominique Challine ◽  
Marie-Hélène Delfau-Larue ◽  
Corinne Haioun ◽  
Dominique Desvaux ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cell ◽  
Dna Sequences ◽  
Primary Effusion Lymphoma ◽  
Polymerase Chain Reaction Amplification ◽  
Human Herpesvirus ◽  
Body Cavity ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Immunodeficiency Virus

Abstract We describe a case of an 87-year-old human immunodeficiency virus (HIV)–negative man who developed a primary pleural lymphoma without any identifiable tumor mass associated with human herpesvirus 8 (HHV-8) infection. A large T-cell lymphoma was diagnosed based on morphologic, immunophenotypic, and molecular findings. The HHV-8 DNA sequences were detected using specific polymerase chain reaction amplification in the lymphomatous effusion. Study of the patient's serum confirmed the HHV-8 infection. This case report displays the characteristic features of HHV-8–related body cavity-based lymphoma/primary effusion lymphoma previously reported in HIV-seronegative patients, except that it is of T-cell origin. Whether this case may be included or not within the primary effusion lymphoma entity, the association of a pleural T-cell non-Hodgkin lymphoma with HHV-8 infection raises the question of the possible occurrence of T cells as the target of malignant transformation associated with HHV-8 infection.

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