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Production and Purification of Baculovirus for Gene Therapy Application
Published on: April 9, 2018
A surface-modified baculovirus vector with improved gene delivery to B-lymphocytic cells
Jing Ge1, Yishu Huang, Xiaotong Hu
1Department Of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
Journal of Biotechnology
|March 22, 2007
Summary
Researchers engineered a baculovirus to target B cells by incorporating an Epstein-Barr virus peptide that binds to CD21. This modified virus, vAc-gp350EGFP, demonstrated enhanced B cell transduction, confirming CD21 interaction as the entry mechanism.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Epstein-Barr virus (EBV) utilizes surface proteins for B-lymphocyte interaction.
- CD21 is a key surface receptor on B-lymphocytes mediating EBV entry.
- Baculovirus vectors are widely used for gene delivery and expression.
Purpose of the Study:
- To enhance baculovirus transduction efficiency in B-lymphocytes.
- To investigate the role of CD21 in baculovirus entry into B cells.
- To develop a targeted viral vector for B cell applications.
Main Methods:
- Insertion of a CD21-binding peptide from EBV gp350/220 into baculovirus gp64.
- Construction and expression of recombinant baculovirus vAc-gp350EGFP.
- Assessment of transduction efficiency in various B and non-B cell lines.
- Inhibition studies using anti-CD21 antibodies.
Main Results:
- Recombinant vAc-gp350EGFP showed significantly increased transduction in multiple B cell lines (Raji, HR1, B95-8, BJAB, DG75).
- Enhanced transduction was specific to B cells, with no increase observed in HEK293 and HeLa non-B cell lines.
- Transduction efficiency in Raji cells exhibited saturation at high multiplicities of infection and was dose-dependently inhibited by anti-CD21 antibody.
Conclusions:
- The engineered baculovirus vAc-gp350EGFP effectively targets B cells via CD21-mediated entry.
- This strategy enhances baculovirus transduction efficiency in B-lymphocytes.
- The findings support the development of CD21-targeted baculovirus vectors for B cell research and therapy.
