Murine retroviral vector producer cells survival and toxicity in the dog liver.
To develop a safe method to target gene delivery into intrahepatic tumors, we examined the toxicity of intrahepatic (IH) injection of retroviral vector producer cells (VPC) into the canine liver. VPC have been demonstrated to effectively transfer genes in vivo. To evaluate for adverse effects form xenogeneic cell transplantation, mongrel dogs were injected IH with 1 x 10(9) murine LTKOSN.2 VPC divided into three aliquots. The animals were then monitored for acute toxicity induced by the VPC. The intraoperative IH injections of the cells were tolerated without difficulty. Starting 7 days after IH injection, the dogs then received intravenous ganciclovir (GCV) twice daily (5 mg/kg) for 7 days. GCV treatment did not cause significant toxicities. Dogs underwent serial blood tests to evaluate bone marrow, renal, liver and immunological function. Complete blood counts, electrolytes, liver function and renal function tests remained normal except for mild elevations of alkaline phosphatase. Histologic examination of liver tissues from the IH injection site revealed no apparent normal tissue destruction induced by the VPC. Two of the four treated dogs underwent liver biopsy on day 3. These biopsy specimens were cultured and persistent, viable VPC were recovered. The dogs mounted an antibody response to the murine VPC that was first demonstrated 5 days post injection. PCR analysis demonstrated low level gene transfer into dog liver tissue. Overall, our results demonstrate that IH xenogeneic VPC injections are not accompanied by significant adverse effects over a 1 month period following administration into the canine liver. These data support the safety aspects of using murine VPC in Phase I clinical trials.
Link
C J
CJ
Human Gene Therapy Research Institute, Iowa Health Systems, 1415 Woodland Ave., Des Moines, IA 50309, USA. chrclink@aol.com
Seregina
T
T
Levy
J P
JP
Martin
M
M
Ackermann
M
M
Moorman
D W
DW
D000469:Alkaline Phosphatase / Q000097:blood; D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D004285:Dogs; D015774:Ganciclovir / Q000494:pharmacology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006403:Hematologic Tests; D018259:Herpesvirus 1, Human / Q000201:enzymology / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000502:physiology; D007677:Kidney Function Tests; D008099:Liver / Q000473:pathology* / Q000502:physiology; D008111:Liver Function Tests; D051379:Mice; D023421:Models, Animal; D016133:Polymerase Chain Reaction; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000637:transplantation
D013937:Thymidine Kinase; D000469:Alkaline Phosphatase; D015774:Ganciclovir