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Establishment and characterization of a primary effusion (body cavity- based) lymphoma cell line (BC-3) harboring kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) in the absence of Epstein-Barr virus

L Arvanitakis, EA Mesri, RG Nador, JW Said, AS Asch, DM Knowles and E Cesarman

Department of Pathology, New York Hospital-Cornell Medical Center, New York 10021, USA.

The recently identified Kaposi's sarcoma-associated herpesvirus (KSHV), also known as human herpesvirus 8 (HHV-8), has been found to be consistently associated with an unusual subset of acquired immunodeficiency syndrome-related lymphomas, the so-called body cavity- based lymphomas (BCBL) or primary effusion lymphomas (PEL). These lymphomas are characterized by a unique spectrum of morphologic and molecular characteristics, and grow as lymphomatous effusions without an identifiable contiguous tumor mass. Until now, efforts to delineate the role of KSHV in the pathogenesis of PELs have been hampered by the lack of appropriate model systems and the concomitant presence of Epstein-Barr virus (EBV) in nearly all cases examined, and in all previously established cell lines. We now report the establishment and characterization of a novel PEL cell line, BC-3, which is KSHV+ by polymerase chain reaction (PCR) but EBV- as assessed by a variety of methods including PCR for EBER, EBNA-2, and EBNA-3C. This cell line was established from a lymphomatous effusion obtained from an HIV- patient, and has immunophenotypic and molecular features consistent with the diagnosis of PEL, including an indeterminate immunophenotype with a B- cell immunogenotype and lack of c-myc proto-oncogene rearrangements. Pulsed-field gel electrophoresis shows an intact KSHV genome of about 170 kb both in the cell line and in the viral isolate, whereas herpesvirus-like capsids are visible by electron microscopy. Consequently, the BC-3 cell line represents an invaluable tool as a source of KSHV, for both the evaluation of the pathogenic potential of this virus and the mechanistic characterization of its role in the development of Kaposi's sarcoma and malignant lymphoma.

Volume 88, Issue 7, pp. 2648-2654, 10/01/1996
Copyright © 1996 by The American Society of Hematology


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Human Herpesvirus 8 K1-Associated Nuclear Factor-kappa B-Dependent Promoter Activity: Role in Kaposi's Sarcoma Inflammation?
J Natl Cancer Inst Monographs, December 1, 2000; 2000(28): 15 - 23.
[Abstract] [Full Text] [PDF]


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J. S. Cannon, D. Ciufo, A. L. Hawkins, C. A. Griffin, M. J. Borowitz, G. S. Hayward, and R. F. Ambinder
A New Primary Effusion Lymphoma-Derived Cell Line Yields a Highly Infectious Kaposi's Sarcoma Herpesvirus-Containing Supernatant
J. Virol., November 1, 2000; 74(21): 10187 - 10193.
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J. Chen, K. Ueda, S. Sakakibara, T. Okuno, and K. Yamanishi
Transcriptional Regulation of the Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor Gene
J. Virol., September 15, 2000; 74(18): 8623 - 8634.
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J. Gen. Virol.Home page
Y.-J. Zhang, J.-H. Deng, C. Rabkin, and S.-J. Gao
Hot-spot variations of Kaposi's sarcoma-associated herpesvirus latent nuclear antigen and application in genotyping by PCR-RFLP
J. Gen. Virol., August 1, 2000; 81(8): 2049 - 2058.
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L. Gradoville, J. Gerlach, E. Grogan, D. Shedd, S. Nikiforow, C. Metroka, and G. Miller
Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 50/Rta Protein Activates the Entire Viral Lytic Cycle in the HH-B2 Primary Effusion Lymphoma Cell Line
J. Virol., July 1, 2000; 74(13): 6207 - 6212.
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J.-G. Judde, V. Lacoste, J. Briere, E. Kassa-Kelembho, E. Clyti, P. Couppie, C. Buchrieser, M. Tulliez, J. Morvan, and A. Gessain
Monoclonality or Oligoclonality of Human Herpesvirus 8 Terminal Repeat Sequences in Kaposi's Sarcoma and Other Diseases
J Natl Cancer Inst, May 3, 2000; 92(9): 729 - 736.
[Abstract] [Full Text] [PDF]


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F. Pica, A. Volpi, A. Serafino, M. Fraschetti, O. Franzese, and E. Garaci
Autocrine nerve growth factor is essential for cell survival and viral maturation in HHV-8-infected primary effusion lymphoma cells
Blood, May 1, 2000; 95(9): 2905 - 2912.
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J. Park, D. Lee, T. Seo, J. Chung, and J. Choe
Kaposi’s sarcoma-associated herpesvirus (human herpesvirus-8) open reading frame 36 protein is a serine protein kinase
J. Gen. Virol., April 1, 2000; 81(4): 1067 - 1071.
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A. Carbone, A. Gloghini, D. Bontempo, P. Monini, U. Tirelli, R. Volpe, P. J. Browning, and G. Gaidano
Proliferation in HHV-8-Positive Primary Effusion Lymphomas Is Associated with Expression of HHV-8 Cyclin but Independent of p27kip1
Am. J. Pathol., April 1, 2000; 156(4): 1209 - 1215.
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M. Haque, J. Chen, K. Ueda, Y. Mori, K. Nakano, Y. Hirata, S. Kanamori, Y. Uchiyama, R. Inagi, T. Okuno, et al.
Identification and Analysis of the K5 Gene of Kaposi's Sarcoma-Associated Herpesvirus
J. Virol., March 15, 2000; 74(6): 2867 - 2875.
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JEMHome page
T.-Y. Yang, S.-C. Chen, M. W. Leach, D. Manfra, B. Homey, M. Wiekowski, L. Sullivan, C.-H. Jenh, S. K. Narula, S. W. Chensue, et al.
Transgenic Expression of the Chemokine Receptor Encoded by Human Herpesvirus 8 Induces an Angioproliferative Disease Resembling Kaposi's Sarcoma
J. Exp. Med., February 7, 2000; 191(3): 445 - 454.
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A. J. Aguirre and E. S. Robertson
Epstein-Barr Virus Recombinants from BC-1 and BC-2 Can Immortalize Human Primary B Lymphocytes with Different Levels of Efficiency and in the Absence of Coinfection by Kaposi's Sarcoma-Associated Herpesvirus
J. Virol., January 15, 2000; 74(2): 735 - 743.
[Abstract] [Full Text] [PDF]


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BloodHome page
Y. Aoki and G. Tosato
Role of Vascular Endothelial Growth Factor/Vascular Permeability Factor in the Pathogenesis of Kaposi's Sarcoma-Associated Herpesvirus-Infected Primary Effusion Lymphomas
Blood, December 15, 1999; 94(12): 4247 - 4254.
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L. J. Bello, A. J. Davison, M. A. Glenn, A. Whitehouse, N. Rethmeier, T. F. Schulz, and J. Barklie Clements
The human herpesvirus-8 ORF 57 gene and its properties
J. Gen. Virol., December 1, 1999; 80(12): 3207 - 3215.
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V. Varthakavi, P. J. Browning, and P. Spearman
Human Immunodeficiency Virus Replication in a Primary Effusion Lymphoma Cell Line Stimulates Lytic-Phase Replication of Kaposi's Sarcoma-Associated Herpesvirus
J. Virol., December 1, 1999; 73(12): 10329 - 10338.
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A. Ibrahimbacha, M. Farah, and J. Saluja
An HIV-Infected Patient With Pleural Effusion
Chest, October 1, 1999; 116(4): 1113 - 1115.
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A. V. Moses, K. N. Fish, R. Ruhl, P. P. Smith, J. G. Strussenberg, L. Zhu, B. Chandran, and J. A. Nelson
Long-Term Infection and Transformation of Dermal Microvascular Endothelial Cells by Human Herpesvirus 8
J. Virol., August 1, 1999; 73(8): 6892 - 6902.
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R. Sadler, L. Wu, B. Forghani, R. Renne, W. Zhong, B. Herndier, and D. Ganem
A Complex Translational Program Generates Multiple Novel Proteins from the Latently Expressed Kaposin (K12) Locus of Kaposi's Sarcoma-Associated Herpesvirus
J. Virol., July 1, 1999; 73(7): 5722 - 5730.
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J. P. Zoeteweij, S. T. Eyes, J. M. Orenstein, T. Kawamura, L. Wu, B. Chandran, B. Forghani, and A. Blauvelt
Identification and Rapid Quantification of Early- and Late-Lytic Human Herpesvirus 8 Infection in Single Cells by Flow Cytometric Analysis: Characterization of Antiherpesvirus Agents
J. Virol., July 1, 1999; 73(7): 5894 - 5902.
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H. Katano, Y. Sato, T. Kurata, S. Mori, and T. Sata
High Expression of HHV-8-Encoded ORF73 Protein in Spindle-Shaped Cells of Kaposi's Sarcoma
Am. J. Pathol., July 1, 1999; 155(1): 47 - 52.
[Abstract] [Full Text] [PDF]


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P. Kellam, D. Bourboulia, N. Dupin, C. Shotton, C. Fisher, S. Talbot, C. Boshoff, and R. A. Weiss
Characterization of Monoclonal Antibodies Raised against the Latent Nuclear Antigen of Human Herpesvirus 8
J. Virol., June 1, 1999; 73(6): 5149 - 5155.
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J.-C. Zong, D. M. Ciufo, D. J. Alcendor, X. Wan, J. Nicholas, P. J. Browning, P. L. Rady, S. K. Tyring, J. M. Orenstein, C. S. Rabkin, et al.
High-Level Variability in the ORF-K1 Membrane Protein Gene at the Left End of the Kaposi's Sarcoma-Associated Herpesvirus Genome Defines Four Major Virus Subtypes and Multiple Variants or Clades in Different Human Populations
J. Virol., May 1, 1999; 73(5): 4156 - 4170.
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H Gruffat, S Portes-Sentis, A Sergeant, and E Manet
Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8) encodes a homologue of the Epstein-Barr virus bZip protein EB1
J. Gen. Virol., March 1, 1999; 80(3): 557 - 561.
[Abstract]


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R. Sun, S.-F. Lin, K. Staskus, L. Gradoville, E. Grogan, A. Haase, and G. Miller
Kinetics of Kaposi's Sarcoma-Associated Herpesvirus Gene Expression
J. Virol., March 1, 1999; 73(3): 2232 - 2242.
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NEJMHome page
J. M. Orenstein, B. Herndier, J. C. Aster, and D. Jones
Appearance of Human Herpesvirus 8 on Electron Microscopy
N. Engl. J. Med., January 7, 1999; 340(1): 62 - 64.
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Y Yu, J. Black, C. Goldsmith, P. Browning, K Bhalla, and M. Offermann
Induction of human herpesvirus-8 DNA replication and transcription by butyrate and TPA in BCBL-1 cells
J. Gen. Virol., January 1, 1999; 80(1): 83 - 90.
[Abstract]


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J. Friborg Jr., W.-P. Kong, C. C. Flowers, S. L. Flowers, Y. Sun, K. E. Foreman, B. J. Nickoloff, and G. J. Nabel
Distinct Biology of Kaposi's Sarcoma-Associated Herpesvirus from Primary Lesions and Body Cavity Lymphomas
J. Virol., December 1, 1998; 72(12): 10073 - 10082.
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A. Matolcsy, R. G. Nador, E. Cesarman, and D. M. Knowles
Immunoglobulin VH Gene Mutational Analysis Suggests that Primary Effusion Lymphomas Derive from Different Stages of B Cell Maturation
Am. J. Pathol., November 1, 1998; 153(5): 1609 - 1614.
[Abstract] [Full Text] [PDF]


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BloodHome page
J. F. Tisdale, A. K. Stewart, B. Dickstein, R. F. Little, I. Dube, D. Cappe, C. E. Dunbar, and K. E. Brown
Molecular and Serological Examination of the Relationship of Human Herpesvirus 8 to Multiple Myeloma: orf 26 Sequences in Bone Marrow Stroma Are Not Restricted to Myeloma Patients and Other Regions of the Genome Are Not Detected
Blood, October 15, 1998; 92(8): 2681 - 2687.
[Abstract] [Full Text] [PDF]


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NEJMHome page
D. Jones, M. E. Ballestas, K. M. Kaye, J. M. Gulizia, G. L. Winters, J. Fletcher, D. T. Scadden, and J. C. Aster
Primary-Effusion Lymphoma and Kaposi's Sarcoma in a Cardiac-Transplant Recipient
N. Engl. J. Med., August 13, 1998; 339(7): 444 - 449.
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M.-S. Raab, J.-C. Albrecht, A. Birkmann, D. Lang, B. Fleckenstein, and F. Neipel
The Immunogenic Glycoprotein gp35-37 of Human Herpesvirus 8 Is Encoded by Open Reading Frame K8.1
J. Virol., August 1, 1998; 72(8): 6725 - 6731.
[Abstract] [Full Text] [PDF]


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BloodHome page
L. G. Chatlynne, W. Lapps, M. Handy, Y. Q. Huang, R. Masood, A. S. Hamilton, J. W. Said, H.P. Koeffler, M. H. Kaplan, A. Friedman-Kien, et al.
Detection and Titration of Human Herpesvirus-8-Specific Antibodies in Sera From Blood Donors, Acquired Immunodeficiency Syndrome Patients, and Kaposi's Sarcoma Patients Using a Whole Virus Enzyme-Linked Immunosorbent Assay
Blood, July 1, 1998; 92(1): 53 - 58.
[Abstract] [Full Text] [PDF]


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N. Regamey, G. Cathomas, M. Schwager, M. Wernli, T. Harr, and P. Erb
High Human Herpesvirus 8 Seroprevalence in the Homosexual Population in Switzerland
J. Clin. Microbiol., June 1, 1998; 36(6): 1784 - 1786.
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R. Renne, D. Blackbourn, D. Whitby, J. Levy, and D. Ganem
Limited Transmission of Kaposi's Sarcoma-Associated Herpesvirus in Cultured Cells
J. Virol., June 1, 1998; 72(6): 5182 - 5188.
[Abstract] [Full Text] [PDF]


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J. A. Reed, R. G. Nador, D. Spaulding, Y. Tani, E. Cesarman, and D. M. Knowles
Demonstration of Kaposi's Sarcoma-Associated Herpes Virus Cyclin D Homolog in Cutaneous Kaposi's Sarcoma by Colorimetric In Situ Hybridization Using a Catalyzed Signal Amplification System
Blood, May 15, 1998; 91(10): 3825 - 3832.
[Abstract] [Full Text] [PDF]


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BloodHome page
C. Boshoff, S.-J. Gao, L. E. Healy, S. Matthews, A. J. Thomas, L. Coignet, R. A. Warnke, J. A. Strauchen, E. Matutes, O. W. Kamel, et al.
Establishing a KSHV+ Cell Line (BCP-1) From Peripheral Blood and Characterizing Its Growth in Nod/SCID Mice
Blood, March 1, 1998; 91(5): 1671 - 1679.
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R. Sarid, O. Flore, R. A. Bohenzky, Y. Chang, and P. S. Moore
Transcription Mapping of the Kaposi's Sarcoma-Associated Herpesvirus (Human Herpesvirus 8) Genome in a Body Cavity-Based Lymphoma Cell Line (BC-1)
J. Virol., February 1, 1998; 72(2): 1005 - 1012.
[Abstract] [Full Text] [PDF]


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BloodHome page
A. Carbone, G. Gaidano, A. Gloghini, L. M. Larocca, D. Capello, V. Canzonieri, A. Antinori, U. Tirelli, B. Falini, and R. Dalla-Favera
Differential Expression of BCL-6, CD138/Syndecan-1, and Epstein-Barr Virus-Encoded Latent Membrane Protein-1 Identifies Distinct Histogenetic Subsets of Acquired Immunodeficiency Syndrome-Related Non-Hodgkin's Lymphomas
Blood, February 1, 1998; 91(3): 747 - 755.
[Abstract] [Full Text] [PDF]


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X. Bai, G. Hosler, B. B. Rogers, D. B. Dawson, and R. H. Scheuermann
Quantitative polymerase chain reaction for human herpesvirus diagnosis and measurement of Epstein–Barr virus burden in posttransplant lymphoproliferative disorder
Clin. Chem., October 1, 1997; 43(10): 1843 - 1849.
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