Kaposi's sarcoma-associated herpesvirus can productively infect primary human keratinocytes and alter their growth properties.

TitleKaposi's sarcoma-associated herpesvirus can productively infect primary human keratinocytes and alter their growth properties.
Publication TypeJournal Article
Year of Publication2001
AuthorsCerimele F, Curreli F, Ely S, Friedman-Kien AE, Cesarman E, Flore O
JournalJ Virol
Date Published2001 Mar
KeywordsAntigens, Viral, Cells, Cultured, Cytokines, Endothelial Growth Factors, Fibroblast Growth Factor 2, Herpesviridae Infections, Herpesvirus 8, Human, Humans, Immunohistochemistry, Keratinocytes, Lymphokines, Nuclear Proteins, Phosphoproteins, Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, Virus Replication

Previous studies have shown the presence of Kaposi's sarcoma-associated herpesvirus (KSHV/HHV8) DNA in endothelial cells, in keratinocytes in the basal layer of the epidermis overlying plaque-stage nodular lesions of cutaneous Kaposi's sarcoma (KS), and in the epithelial cells of eccrine glands within KS lesions. We infected primary cell cultures of human keratinocytes with KSHV/HHV8. At 6 days post infection, transcription of viral genes was detected by reverse transcriptase PCR (RT-PCR), and protein expression was documented by an immunofluorescence assay with an anti-LANA monoclonal antibody. To determine whether the viral lytic cycle was inducible by chemical treatment, KSHV/HHV8-infected keratinocytes were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) and RT-PCR was performed to confirm the transcription of lytic genes such as open reading frame 26, (which encodes a capsid protein). Finally, to assess infectious viral production, other primary human cells (human umbilical vein endothelial cells), were infected with concentrated supernatant of KSHV-infected, TPA-induced keratinocytes and the presence of viral transcripts was confirmed by RT-PCR. The uninfected keratinocytes senesced 3 to 5 weeks after mock infection, while the KSHV/HHV8-infected keratinocytes continued to proliferate and to date are still in culture. However, 8 weeks after infection, viral genomes were no longer detectable by nested PCR. Although the previously KSHV/HHV8-infected keratinocytes still expressed epithelial markers, they acquired new characteristics such as contact inhibition loss, telomerase activity, anchorage-independent growth, and changes in cytokine production. These results show that KSHV/HHV8, like other herpesviruses, can infect and replicate in epithelial cells in vitro and suggest that in vivo these cells may play a significant role in the establishment of KSHV/HHV8 infection and viral transmission.

Alternate JournalJ Virol
PubMed ID11160746
PubMed Central IDPMC114826
Related Faculty: 
Ethel Cesarman, M.D., Ph.D.

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