Optimized lentiviral vector production and purification procedure prevents immune response after transduction of mouse brain
- 6 October 2003
- journal article
- Published by Springer Nature in Gene Therapy
- Vol. 10 (23) , 1933-1940
- https://doi.org/10.1038/sj.gt.3302094
Abstract
HIV-derived lentiviral vectors are efficient vehicula to deliver genes into the brain and hold great promise for future gene therapy of neurodegenerative disorders. However, administration of the current vector preparations in mouse brain was found to induce a systemic immune response to vector proteins and a modest inflammation in the brain. Moreover, serum antibodies from vector-treated animals were capable of partially neutralizing lentiviral vector-mediated transduction in cell culture. To avoid this unexpected immune reaction, we have optimized new vector production and purification protocols. Purification by sucrose gradient ultracentrifugation abolished the immune response, but vector titers also decreased substantially. Lentiviral vector production in the absence of serum in the cell culture medium equally reduced immunogenicity without affecting transduction efficiency. These results have important implications for future clinical use of lentiviral vectors, and for the use of lentiviral vectors to create animal models for neurodegenerative diseases that have an important neuroinflammatory component.Keywords
This publication has 27 references indexed in Scilit:
- Immune Response to Fetal Calf Serum by Two Adenosine Deaminase-Deficient Patients After T Cell Gene TherapyHuman Gene Therapy, 2002
- Characterization of Lentiviral Vector-Mediated Gene Transfer in Adult Mouse BrainHuman Gene Therapy, 2002
- Neuroprotective Effect of a CNTF-Expressing Lentiviral Vector in the Quinolinic Acid Rat Model of Huntington's DiseaseNeurobiology of Disease, 2001
- In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected miceNature Medicine, 2001
- VSV-G Pseudotyped Lentiviral Vector Particles Produced in Human Cells Are Inactivated by Human SerumMolecular Therapy, 2000
- Complete and Long-Term Rescue of Lesioned Adult Motoneurons by Lentiviral-Mediated Expression of Glial Cell Line-Derived Neurotrophic Factor in the Facial NucleusJournal of Neuroscience, 2000
- Lentiviral Vectors as a Gene Delivery System in the Mouse Midbrain: Cellular and Behavioral Improvements in a 6-OHDA Model of Parkinson's Disease Using GDNFExperimental Neurology, 2000
- Minimal exclusion of plasma membrane proteins during retroviral envelope formationProceedings of the National Academy of Sciences, 2000
- In Vivo Protection of Nigral Dopamine Neurons by Lentiviral Gene Transfer of the Novel GDNF-Family Member Neublastin/ArteminMolecular and Cellular Neuroscience, 2000
- Chronic brain inflammation and persistent herpes simplex virus 1 thymidine kinase expression in survivors of syngeneic glioma treated by adenovirus-mediated gene therapy: Implications for clinical trialsNature Medicine, 1999