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Related Experiment Video

Updated: Dec 31, 2025

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Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector.

Aurelien B L Colamartino1,2, William Lemieux1,2, Panojot Bifsha2

  • 1Department of Microbiology, Infectiology and Immunology, University of Montréal, Montréal, QC, Canada.

Frontiers in Immunology
|January 11, 2020
PubMed
Summary

Baboon envelope pseudotyped lentiviral vectors (BaEV-LVs) enhance natural killer (NK) cell transduction efficiency for immunotherapy. BaEV-LVs successfully transduced NK cells, enabling CAR expression and specific cancer cell killing.

Keywords:
NK amplification and expansion system (NKAES)NK-cell transductionbaboon retrovirus envelope pseudotyped lentivectorschimeric antigen receptorcytotoxicity

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Area of Science:

  • Immunology
  • Gene Therapy
  • Cell Biology

Background:

  • Natural killer (NK) cell resistance to genetic modification poses a significant challenge for NK cell immunotherapy development.
  • Efficient gene delivery into NK cells is crucial for enhancing their therapeutic potential and enabling advanced applications like CAR-based therapies.

Purpose of the Study:

  • To evaluate the efficacy of Baboon envelope pseudotyped lentiviral vectors (BaEV-LVs) for transducing human NK cells.
  • To compare BaEV-LV transduction efficiency against other lentiviral vector pseudotypes.
  • To assess the functional capacity of BaEV-LV transduced NK cells expressing chimeric antigen receptors (CARs).

Main Methods:

  • Human NK cells, including freshly isolated (FI-NK) and expanded (NKAES) populations, were transduced with BaEV-LVs encoding eGFP.
  • Transduction efficiency was quantified and compared with Vesicular Stomatitis Virus type-G (VSV-G), RD114, and Measles Virus (MV) pseudotyped lentiviral vectors.
  • NK cells were engineered to express CARs (CAR-CD22 and dual CD19/CD22-CAR) via BaEV-LV transduction, and their expression and cytotoxic activity were evaluated against target cell lines.

Main Results:

  • BaEV-LVs achieved significantly higher transduction rates in NKAES (83.4%) compared to FI-NK (23.0%) and other pseudotypes (p < 0.0001).
  • Sustained transgene expression was observed for at least 21 days post-transduction.
  • BaEV-LV mediated CAR-CD22 expression in NKAES cells resulted in specific killing of the NK-resistant RS4;11 pre-B acute lymphoblastic leukemia cell line.
  • Dual-CAR-NK cells effectively killed both CD19 and CD22 knockout RS4;11 cells, demonstrating broad targeting capabilities.

Conclusions:

  • BaEV-LVs represent a highly efficient and promising tool for genetic modification of human NK cells, overcoming previous transduction hurdles.
  • This advanced vector system facilitates the development of potent NK cell-based immunotherapies, including those utilizing CAR constructs.
  • BaEV-LVs hold significant potential for advancing NK cell research and clinical translation of NK cell-based cancer treatments.