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This Article Full Text (Papers in Press[PDF]) All Versions of this Article: vir.0.011650-0v1 90/8/1943    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Work, T. M Articles by Casey, J. W Search for Related Content PubMed PubMed Citation Articles by Work, T. M Articles by Casey, J. W Agricola Articles by Work, T. M Articles by Casey, J. W

In vitro biology of fibropapilloma-associated turtle herpesvirus and host cells in Hawaiian green turtles (Chelonia mydas).

¹ US Geological Survey-NWHC-HFS;
² NOAA-NMFS-PIFSC;
³ US Geological Survey NWHC-HFS;
⁴ University of Hawaii HIMB;
⁵ Cornell University

⁶ E-mail: thierry_work{at}usgs.gov

Fibropapillomatosis (FP) of green turtles has a global distribution and causes debilitating tumors of the skin and internal organs in several species of marine turtles. FP is associated with a presently non-cultivable alphaherpesvirus (Chelonid fibropapilloma-associated herpesvirus-CFPHV). Our aims were to employ quantitative PCR (qPCR) targeted to pol DNA of CFPHV to determine 1) if DNA sequesters by tumor size and/or cell type, 2) whether subculturing of cells is a viable strategy for isolating CFPHV and 3) whether CFPHV can be induced to a lytic growth cycle in vitro using chemical modulators of replication (CMR), temperature variation, or co-cultivation. Additional objectives included determining whether non tumored and tumored cells behave differently in vitro and confirming the phenotype of cultured cells using cell type-specific antigens. CFPHV pol DNA was preferentially concentrated in dermal fibroblasts of skin tumors, and amount of viral DNA per cell was independent of tumor size. Copy number of CFPHV pol DNA per cell rapidly decreased with cell doubling of tumor-derived fibroblasts in culture. Attempts to induce viral replication in known CFPHV virus DNA-positive cells using temperature or CMR failed. No significant differences were seen in in vitro morphology or growth characteristics of fibroblasts from tumored and paired normal skin nor from CFPHV pol DNA positive intestinal tumor cells. Tumor cells were confirmed as fibroblasts or keratinocytes by positive staining with anti-vimentin and anti-pancytokeratin antibodies, respectively. CFPHV continues to be refractory to in vitro cultivation.

Received 9 March 2009; accepted 21 April 2009.