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Journal of Virology, July 2003, p. 8147-8152, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.8147-8152.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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

Naoki Inoue,1* Jörn Winter,1,2 Renu B. Lal,2 Margaret K. Offermann,3 and Shin Koyano1,{dagger}

Division of Viral and Rickettsial Diseases,1 Division of AIDS, STD, TB and Laboratory Research, Centers for Disease Control and Prevention,2 Winship Cancer Center, Emory University, Atlanta, Georgia3

Received 4 February 2003/ Accepted 25 April 2003

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.

* Corresponding author. Mailing address: Mailstop G18, Herpesvirus Section, DVRD, CDC, 1600 Clifton Rd., Atlanta, GA 30333. Phone: (404) 639-4219. Fax: (404) 639-0049. E-mail: nai0{at}cdc.gov.

{dagger} Present address: Department of Pediatrics, Asahikawa Medical College, Japan.

Journal of Virology, July 2003, p. 8147-8152, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.8147-8152.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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