Document Detail


In vitro biology of fibropapilloma-associated turtle herpesvirus and host cells in Hawaiian green turtles (Chelonia mydas).
MedLine Citation:
PMID:  19386781     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fibropapillomatosis (FP) of green turtles has a global distribution and causes debilitating tumours 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 targeted to pol DNA of CFPHV to determine (i) if DNA sequesters by tumour size and/or cell type, (ii) whether subculturing of cells is a viable strategy for isolating CFPHV and (iii) whether CFPHV can be induced to a lytic growth cycle in vitro using chemical modulators of replication (CMRs), temperature variation or co-cultivation. Additional objectives included determining whether non-tumour and tumour 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 tumours and the amount of viral DNA per cell was independent of tumour size. Copy number of CFPHV pol DNA per cell rapidly decreased with cell doubling of tumour-derived fibroblasts in culture. Attempts to induce viral replication in known CFPHV-DNA-positive cells using temperature or CMR failed. No significant differences were seen in in vitro morphology or growth characteristics of fibroblasts from tumour cells and paired normal skin, nor from CFPHV pol-DNA-positive intestinal tumour cells. Tumour 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.
Authors:
Thierry M Work; Julie Dagenais; George H Balazs; Joanne Schumacher; Teresa D Lewis; Jo-Ann C Leong; Rufina N Casey; James W Casey
Publication Detail:
Type:  Journal Article     Date:  2009-04-22
Journal Detail:
Title:  The Journal of general virology     Volume:  90     ISSN:  0022-1317     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-07-13     Completed Date:  2009-09-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1943-50     Citation Subset:  IM    
Affiliation:
US Geological Survey, National Wildlife Health Center, Honolulu Field Station, Honolulu, HI 96850, USA. thierry_work@usgs.gov
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MeSH Terms
Descriptor/Qualifier:
Alphaherpesvirinae / growth & development,  isolation & purification*
Animals
Cells, Cultured
Coculture Techniques / methods
Fibroblasts / virology
Hawaii
Herpesviridae Infections / veterinary*,  virology
Keratinocytes / virology
Papilloma / veterinary*,  virology
Turtles / virology*
Virus Activation

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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