Human cytomegalovirus infection in a retinoblastoma cell line in vitro.
BACKGROUND
The focus of these studies was to determine whether the Y79 human retinoblastoma cell line could function as a good in vitro model system for studying human cytomegalovirus (HCMV) infection.


METHODS
Y79 cells were exposed to an HCMV mutant carrying a LacZ gene, and the resulting beta-galactosidase expression in infected cells was assessed by flow cytometry. The extent to which the three classes of viral gene products immediate early, early, and late proteins - were expressed was analyzed by immunohistochemical staining and Western blotting. Infected Y79 cells were also co-cultivated on human foreskin fibroblast (SF cell) cultures to recover virus.


RESULTS
Infection of Y79 cells with the virus resulted in beta-galactosidase expression as detected by flow-cytometric analysis. Immunohistochemical staining revealed that a portion of Y79 cells expressed antigens reactive to monoclonal antibodies against immediate early, early, and late HCMV proteins. The 43-kDa early gene product was also detected by Western blotting. Infected Y79 cells co-cultivated on SF cell cultures yielded infectious foci, which turned blue following X-gal staining, demonstrating productive HCMV infection in the Y79 cells.


CONCLUSIONS
These results demonstrate that while HCMV can productively infect Y79 cultures, it does so in a highly inefficient manner, leading these authors to conclude that this cell line does not provide a particularly good model system to study HCMV infection.
Kumano
Y
Y
Department of Ophthalmology and Visual Sciences, Washington University Medical Center, St. Louis, MO 63110, USA.
Laycock
K A
KA
Hook
K K
KK
Pepose
J S
JS
Stuart
P M
PM
D015153:Blotting, Western; D003587:Cytomegalovirus / Q000502:physiology*; D005434:Flow Cytometry; D005500:Follow-Up Studies; D005697:Galactosides / Q000096:biosynthesis / Q000235:genetics; D017781:Genes, Immediate-Early / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D007763:Lac Operon / Q000502:physiology; D012367:RNA, Viral / Q000032:analysis; D019572:Retinal Neoplasms / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D012175:Retinoblastoma / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D014407:Tumor Cells, Cultured / Q000821:virology; D014764:Viral Proteins / Q000032:analysis
D005697:Galactosides; D012367:RNA, Viral; D014764:Viral Proteins; C007816:beta-galactoside