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Updated: Dec 30, 2025
Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies
Published on: February 15, 2021
Rapid isolation and enrichment of mouse NK cells for experimental purposes
Maite Alvarez1, Maria C Ochoa2, Luna Minute3
1Program of Immunology and Immunotherapy, CIMA Universidad de Navarra, Pamplona, Spain; Navarra Institute for Health Research (IDISNA), Pamplona, Spain; Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain.
Abstract:
Natural killer (NK) cells have shown to play a critical, but as yet poorly defined, role in the process by which the immune system controls tumor progression. Indeed, NK cell-based immunotherapy, particularly NK cell adoptive transfer therapy, has become a very attractive cancer weapon against multiple types of cancers such as metastatic and hematological cancers. Unfortunately, the implementation of these therapies has been challenged by the existence of immunosuppression mechanisms that have prevented NK cell functionality. Additionally, the development of protocols to obtain purified and functional NK cells has faced some difficulties due to the limitations in the numbers of cells that can be obtained and the development of an exhaustion phenotype with impaired proliferative and functional capabilities during lengthy ex vivo NK cell expansion protocols. Thus, the development of new strategies to obtain a rapid expansion of highly functional NK cells without the appearance of exhaustion is still much needed. This is particularly true in the case of mouse NK cells, a surrogate commonly used to evaluate NK cell biology and human NK cell-based immunotherapeutic alternatives. Here, we describe a feasible and rapid protocol to produce strongly activated mouse NK cells in vivo taking advantage of the hydrodynamic delivery of a plasmid that contains interleukin-15, a cytokine known to cause NK cell expansion and activation, fused with the binding domain of the IL-15Rα ("sushi" domain) and apolipoprotein A-I.
Insights
This study presents a novel method for rapidly expanding functional natural killer (NK) cells in vivo. This breakthrough addresses challenges in cancer immunotherapy by overcoming NK cell exhaustion and improving cell numbers for therapeutic applications.
Area of Science:
- Immunology
- Cancer Biology
- Biotechnology
Background:
- Natural killer (NK) cells are crucial for tumor control and NK cell-based immunotherapy shows promise against various cancers.
- Current challenges include immunosuppression, limited cell numbers, and NK cell exhaustion during ex vivo expansion, hindering therapeutic efficacy.
- Developing rapid expansion protocols for functional NK cells, especially in mouse models, is essential for advancing human immunotherapies.
Purpose of the Study:
- To develop a feasible and rapid protocol for producing strongly activated mouse NK cells in vivo.
- To overcome limitations in current NK cell expansion methods, particularly the development of exhaustion phenotypes.
- To provide a valuable tool for studying NK cell biology and evaluating human NK cell-based immunotherapeutic strategies.
Main Methods:
- Utilized hydrodynamic delivery of a plasmid encoding interleukin-15 (IL-15) fused with the IL-15 receptor alpha (IL-15Rα) 'sushi' domain and apolipoprotein A-I.
- Focused on in vivo expansion and activation of NK cells in a mouse model.
- Evaluated the expansion, activation, and functional potential of the generated NK cells.
Main Results:
- Successfully achieved rapid expansion of strongly activated mouse NK cells in vivo.
- The developed protocol circumvents the need for lengthy ex vivo expansion, mitigating NK cell exhaustion.
- Demonstrated a feasible method for generating a substantial number of functional NK cells.
Conclusions:
- The described hydrodynamic delivery of engineered IL-15 offers a rapid and effective strategy for in vivo NK cell expansion and activation.
- This approach holds significant potential for improving NK cell-based cancer immunotherapies by providing a readily available source of functional NK cells.
- This method provides a valuable preclinical tool for advancing NK cell research and therapeutic development.
