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Capillary endothelial cell tropism of PVC-211 murine leukemia virus and its application for gene transduction
M Masuda1, C A Hanson, N V Dugger
1Laboratory of Molecular Oncology, National Cancer Institute, Frederick, Maryland 21702, USA.
Abstract:
PVC-211 murine leukemia virus (MuLV) causes neurodegenerative disease following inoculation of neonatal, but not adult, mice and rats. It was previously shown that tropism for brain capillary endothelial cells (CEC) was a determinant of the viral neuropathogenicity. In this study, we demonstrate that host age-dependent replication of PVC-211 MuLV in vivo occurs in CEC in the brain as well as in other organs, such as the liver, kidney, and heart. In contrast, primary explant cultures of CEC derived from brains and livers of adult and neonatal rats could be infected by PVC-211 MuLV, suggesting that the age-dependent susceptibility was abrogated in vitro. Although CEC were generally less susceptible to MuLV-mediated gene transduction than fibroblasts, treatment of CEC with 2-deoxyglucose followed by inoculation of a PVC-211 MuLV-pseudotyped vector in the absence of heparin improved the transduction efficiency. These observations support the possibility that PVC-211 MuLV may be useful for establishing models of CEC gene transduction.
Insights
Murine leukemia virus (MuLV) replication depends on host age, affecting brain capillary endothelial cells (CEC) and other organs. In vitro studies suggest age-dependent susceptibility is lost, but CEC gene transduction may be improved.
Area of Science:
- Virology
- Neuroscience
- Cell Biology
Background:
- PVC-211 murine leukemia virus (MuLV) causes neurodegeneration in young animals but not adults.
- Tropism for brain capillary endothelial cells (CEC) is a key factor in MuLV neuropathogenicity.
Purpose of the Study:
- To investigate the host age-dependent replication of PVC-211 MuLV in vivo.
- To explore the susceptibility of primary CEC cultures to MuLV infection and gene transduction in vitro.
- To identify methods for improving MuLV-mediated gene transduction in CEC.
Main Methods:
- In vivo replication studies of PVC-211 MuLV in neonatal and adult mice and rats.
- In vitro infection of primary CEC cultures from neonatal and adult rat brains and livers.
- MuLV-pseudotyped vector-mediated gene transduction of CEC, with and without 2-deoxyglucose and heparin treatment.
Main Results:
- PVC-211 MuLV replicated in CEC and other organs (liver, kidney, heart) in an age-dependent manner in vivo.
- Primary CEC cultures from both neonatal and adult rats were susceptible to MuLV infection, indicating abrogation of age-dependent susceptibility in vitro.
- CEC were generally less susceptible to MuLV gene transduction than fibroblasts, but treatment with 2-deoxyglucose improved transduction efficiency.
Conclusions:
- Host age influences PVC-211 MuLV replication in vivo, particularly in brain capillary endothelial cells.
- In vitro models using primary CEC cultures may not fully recapitulate in vivo age-dependent susceptibility.
- Optimized conditions, such as 2-deoxyglucose treatment, show potential for enhancing MuLV-based gene transduction in CEC.