title: Infection and transmission dynamics of rKSHV.219 in primary endothelial cells.
abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) is the aetiologic agent of Kaposi's sarcoma (KS), a tumour of endothelial cell origin. The study of KS development was aided by the generation of a recombinant GFP (latent)/RFP (lytic)-expressing KSHV (rKSHV.219) by Vieira and O'Hearn (2004). In this study the first data characterising primary endothelial cell infection and transmission with this virus is presented. Infection was predominantly latent and the percentage of GFP-positive cells increased over time. Neither horizontal transmission of infection, nor cellular proliferation, explained this increase. Analysis of latency-associated nuclear antigen (LANA-1) expression revealed that a threshold level of infection was required for GFP expression early post infection. At later time points GFP correlated more closely with LANA-1 expression, likely due to the accumulation of GFP over time. This study provides methodological guidance for the use of rKSHV.21. In addition, it highlights potential problems associated with the use of fluorescent proteins as markers of viral infection.
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, B15 2TT, UK.
authors.forename: Hannah C
authors.initials: HC
authors.lastname: Jeffery
authors.forename: Rachel L
authors.initials: RL
authors.lastname: Wheat
authors.forename: David J
authors.initials: DJ
authors.lastname: Blackbourn
authors.forename: Gerard B
authors.initials: GB
authors.lastname: Nash
authors.forename: Lynn M
authors.initials: LM
authors.lastname: Butler
mesh_terms: D002478:Cells, Cultured; D042783:Endothelial Cells / Q000821:virology*; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D019288:Herpesvirus 8, Human / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D014773:Virology / Q000379:methods; D017735:Virus Latency; D014779:Virus Replication
chemical_list: D049452:Green Fluorescent Proteins
keywords: Endothelial cell; Kaposi's sarcoma-associated herpes virus; Rkshv.219