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Retroviral Transduction of T-cell Receptors in Mouse T-cells
Published on: October 22, 2010
Expression of lymphotoxin gene inserted into Theiler's murine encephalomyelitis virus
M Obuchi1, T Odagiri, H Lizuka
1Department of Microbiology, Kanazawa Medical University, Uchinada, Ishikawa, Japan.
Microbiology and Immunology
|April 1, 1999
Summary
Theiler
Area of Science:
- Neurovirology
- Molecular Biology
- Gene Therapy
Background:
- Theiler's murine encephalomyelitis virus (TMEV) DA strain causes polioencephalomyelitis and spinal cord demyelination.
- TMEV exhibits viral persistence within the central nervous system.
Purpose of the Study:
- To evaluate the potential of TMEV DA strain as a vector for gene delivery into the central nervous system.
- To investigate the expression and biological activity of a foreign gene delivered by the TMEV vector.
Main Methods:
- Genetic engineering of the TMEV DA genome to insert the human lymphotoxin (LT) gene.
- Infection of BHK-21 cells with the recombinant virus.
- Detection of LT expression using immunoblot and ELISA.
- Assessment of LT cytotoxicity on L-929 cells.
Main Results:
- Successful insertion and expression of the human lymphotoxin (LT) gene in the TMEV genome.
- Demonstrated LT expression in infected BHK-21 cells via immunoblot and ELISA.
- Confirmed cytotoxic activity of the expressed LT in L-929 cells.
Conclusions:
- The TMEV DA strain can be engineered as a vector for delivering foreign genes into the central nervous system.
- The recombinant virus expresses functional human lymphotoxin with demonstrated cytotoxic effects.
- This approach holds potential for future gene therapy strategies targeting the central nervous system.
