Retroviral gene transfer into primary human NK cells activated by IL-2 and K562 feeder cells expressing membrane-bound IL-21

Maria A Streltsova; Eugene Barsov; Sofia A Erokhina; Elena I Kovalenko

doi:10.1016/j.jim.2017.08.003

Retroviral gene transfer into primary human NK cells activated by IL-2 and K562 feeder cells expressing membrane-bound IL-21

J Immunol Methods. 2017 Nov:450:90-94. doi: 10.1016/j.jim.2017.08.003. Epub 2017 Aug 10.

Authors

Maria A Streltsova¹, Eugene Barsov², Sofia A Erokhina³, Elena I Kovalenko⁴

Affiliations

¹ Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow 117997, Russia. Electronic address: mstreltsova@mail.ru.
² Cell Design Labs, Inc., Emeryville, CA, USA.
³ Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow 117997, Russia.
⁴ Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow 117997, Russia. Electronic address: lenkovalen@mail.ru.

PMID: 28802832
DOI: 10.1016/j.jim.2017.08.003

Abstract

Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21. The activated NK cells were in less differentiated state and expressed NK cell activation receptors NKG2D, NKp30, CD16, and were highly HLA-DR-positive. This NK cell population was highly susceptible to the transduction by both GFP- and NGFR-expressing retroviral vectors, with transduction efficiency exceeding 50%. More mature CD57⁺ NK cell population was generally resistant to retroviral vector transduction because of poor response to the stimulation. Our findings may facilitate retroviral vector-mediated genetic engineering of human primary NK cells for future immunotherapies.

Keywords: K562-mbIL21 feeder cells; NK cells; Retroviral particles; Transduction.

MeSH terms

CD57 Antigens / immunology
CD57 Antigens / metabolism
Cell Differentiation
Coculture Techniques
Feeder Cells
GPI-Linked Proteins / immunology
GPI-Linked Proteins / metabolism
Genetic Vectors*
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
HLA-DR Antigens / immunology
HLA-DR Antigens / metabolism
Humans
Immunotherapy / methods*
Interleukin-2 / immunology
Interleukin-2 / metabolism*
Interleukins / immunology
Interleukins / metabolism*
Jurkat Cells
K562 Cells
Killer Cells, Natural / immunology
Killer Cells, Natural / metabolism*
Leukemia, Erythroblastic, Acute / genetics
Leukemia, Erythroblastic, Acute / immunology
Leukemia, Erythroblastic, Acute / metabolism
Leukemia, Erythroblastic, Acute / therapy*
Lymphocyte Activation*
NK Cell Lectin-Like Receptor Subfamily K / immunology
NK Cell Lectin-Like Receptor Subfamily K / metabolism
Natural Cytotoxicity Triggering Receptor 3 / immunology
Natural Cytotoxicity Triggering Receptor 3 / metabolism
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism
Receptors, IgG / immunology
Receptors, IgG / metabolism
Receptors, Nerve Growth Factor / genetics
Receptors, Nerve Growth Factor / metabolism
Retroviridae / genetics*
Transduction, Genetic*
Transfection / methods*

Substances

CD57 Antigens
FCGR3B protein, human
GPI-Linked Proteins
HLA-DR Antigens
Interleukin-2
Interleukins
KLRK1 protein, human
NCR3 protein, human
NGFR protein, human
NK Cell Lectin-Like Receptor Subfamily K
Natural Cytotoxicity Triggering Receptor 3
Nerve Tissue Proteins
Receptors, IgG
Receptors, Nerve Growth Factor
Green Fluorescent Proteins
interleukin-21