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Updated: May 31, 2026
Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells
Published on: January 15, 2018
Efficient NK cell transduction with VSV-G-pseudotyped lentiviral vectors
Emmi Järvelä1,2, Jan Koski1, Farhana Jahan1
1iCell Group, Research and Development, Finnish Red Cross Blood Service, 00290 Helsinki, Finland.
Abstract:
The need for safe, allogeneic cell therapies for cancer is driving a growing interest in CAR-NK-based therapies, which, unlike CAR-T cell therapies, offer the potential for off-the-shelf administration. Lentiviruses pseudotyped with vesicular stomatitis virus glycoprotein G (VSV-G) are commonly used for genetic modification of cell therapy products. Their use in NK cells, however, is limited by low transduction efficiency. This study explores the complexities of NK cell transduction using lentiviral vectors pseudotyped with VSV-G. We demonstrate that efficient transduction depends on multiple factors, such as NK cell activation, domain organization of CAR construct, lentivirus pseudotype selection, and the use of transduction enhancers. By optimizing these elements, we achieved effective transduction, facilitating the use of VSV-G-pseudotyped lentiviral vectors for therapeutic NK cell production. Our optimized workflow comprises NK cell activation with interleukins, followed by transduction with an NK-cell-specific CAR construct using VSV-G-pseudotyped LVs in the presence of BX795 and retronectin, resulting in excellent transduction efficiency (91% using GFP and 80% using CAR) without compromising NK cell phenotype, growth, or cytotoxicity. This allows for the use of a widely used gene transfer vector with an excellent safety record for producing therapeutic NK cell products.
Insights
Optimizing lentiviral vector transduction in Natural Killer (NK) cells enhances CAR-NK therapy. This study details a workflow using specific activators and enhancers for efficient, safe NK cell modification for cancer treatment.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR)-NK cell therapy offers potential for off-the-shelf cancer treatment, overcoming limitations of CAR-T cell therapies.
- Lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G (VSV-G) are common for cell modification but show low transduction efficiency in NK cells.
Purpose of the Study:
- To investigate and optimize the factors influencing lentiviral transduction efficiency in Natural Killer (NK) cells.
- To develop a robust workflow for efficient genetic modification of NK cells for therapeutic applications.
Main Methods:
- Exploration of NK cell activation strategies using interleukins.
- Optimization of CAR construct design and lentivirus pseudotype selection.
- Evaluation of transduction enhancers, including BX795 and retronectin, for enhancing VSV-G lentiviral vector transduction.
Main Results:
- Achieved high transduction efficiency (91% GFP, 80% CAR) using an optimized workflow involving IL-activated NK cells, a specific CAR construct, and VSV-G pseudotyped lentiviral vectors with BX795 and retronectin.
- Demonstrated that optimized transduction conditions do not compromise NK cell phenotype, growth, or cytotoxic function.
- Validated the use of VSV-G pseudotyped lentiviral vectors for efficient therapeutic NK cell production.
Conclusions:
- The study presents an optimized protocol for efficient and safe lentiviral transduction of NK cells.
- This workflow facilitates the production of CAR-NK cell products using established lentiviral vector technology with a strong safety profile.
- The findings support the advancement of allogeneic CAR-NK cell therapies for cancer treatment.
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