Mutant herpes simplex virus-mediated suppression of retinoblastoma.
OBJECTIVE
To test the ability of a mutant herpes simplex virus (HSV) hrR3 to inhibit growth of Y79 human retinoblastoma in vitro and in vivo.


METHODS
Cultured Y79 cells were infected with multiplicities of infection (MOI) ranging from 0.004 to 0.1 of hrR3. Surviving cells were counted using trypan blue dye exclusion. Using X-gal staining, expression of the lacZ gene was examined in vitro on day 3 postinfection to evaluate viral replication. Nude mice harboring Y79 tumors subcutaneously received an intraneoplasmic injection of 5 x 10(7) plaque-forming units of hrR3. The tumor sizes were measured weekly. Expression of the lacZ gene was also examined on one week postinfection.


RESULTS
There are 31% and 13% cells surviving in cultured Y79 cells infected by hrR3 at an MOI of 0.1 on days 3 and 5 postinfection respectively compared to those of mock-infected cells. Also more than 70% of Y79 cells were stained with X-gal at an MOI of 0.1 which demonstrated active viral replication in vitro. Virus-treated subcutaneous tumors were smaller than control tumors (p<<0.05, Student's t-test) on days 14, 21, and 28 postinfection. Positive X-gal staining was also observed in the tumor nodule which was challenged with this viral vector.


CONCLUSIONS
We have demonstrated that hrR3 is capable of inhibiting Y79 tumor growth both in cell culture and in nude mice. These data suggest that gene therapy using this mutant HSV vector can be a new supplementary therapeutic modality for retinoblastoma.
Kogishi
J
J
Department of Ophthalmology and Visual Science, Kyoto University Graduate School of Medicine, Japan.
Miyatake
S
S
Hangai
M
M
Akimoto
M
M
Okazaki
K
K
Honda
Y
Y
D000818:Animals; D002470:Cell Survival / Q000502:physiology; D006651:Histocytochemistry; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009154:Mutation / Q000502:physiology*; D019572:Retinal Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000821:virology*; D012175:Retinoblastoma / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D018139:Simplexvirus / Q000235:genetics*; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000378:metabolism
D001616:beta-Galactosidase