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Expansion of cytotoxic natural killer cells in multiple myeloma patients using K562 cells expressing OX40 ligand and membrane-bound IL-18 and IL-21

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Abstract

Background

Natural killer (NK) cell-based immunotherapy is a promising treatment approach for multiple myeloma (MM), but obtaining a sufficient number of activated NK cells remains challenging. Here, we report an improved method to generate ex vivo expanded NK (eNK) cells from MM patients based on genetic engineering of K562 cells to express OX40 ligand and membrane-bound (mb) IL-18 and IL-21.

Methods

K562-OX40L-mbIL-18/-21 cells were generated by transducing K562-OX40L cells with a lentiviral vector encoding mbIL-18 and mbIL-21, and these were used as feeder cells to expand NK cells from peripheral blood mononuclear cells of healthy donors (HDs) and MM patients in the presence of IL-2/IL-15. Purity, expansion rate, receptor expression, and functions of eNK cells were determined over four weeks of culture.

Results

NK cell expansion was enhanced by short exposure of soluble IL-18 and IL-21 with K562-OX40L cells. Co-culture of NK cells with K562-OX40L-mbIL-18/-21 cells resulted in remarkable expansion of NK cells from HDs (9,860-fold) and MM patients (4,929-fold) over the 28-day culture period. Moreover, eNK cells showed increased expression of major activation markers and enhanced cytotoxicity towards target K562, U266, and RPMI8226 cells.

Conclusions

Our data suggest that genetically engineered K562 cells expressing OX40L, mbIL-18, and mbIL-21 improve the expansion of NK cells, increase activation signals, and enhance their cytolytic activity towards MM cells.

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Funding

This study was supported by grants from the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education, Science and Technology (2018R1A2B6006200, 2020R1A2C2010098).

Author information

Author notes

  1. JayaLakshmi Thangaraj, Minh-TrangThi Phan, SoonHo Kweon authors contributed equally to this work

Authors and Affiliations

  1. Research Center for Cancer Immunotherapy, Gwangju, South Korea

    Jaya Lakshmi Thangaraj, Manh-Cuong Vo, Tan-Huy Chu, Ga-Young Song, Seo-Yeon Ahn, Sung-Hoon Jung & Je-Jung Lee

  2. Department of Hematology-Oncology, Chonnam National University Hwasun Hospital, 322 Seoyangro, Hwasun, Jeollanamdo, 519-763, Republic of Korea

    Jaya Lakshmi Thangaraj, Manh-Cuong Vo, Ga-Young Song, Seo-Yeon Ahn, Sung-Hoon Jung, Hyeoung-Joon Kim & Je-Jung Lee

  3. Department of Molecular Medicine, Chonnam National University, Gwangju, South Korea

    Jaya Lakshmi Thangaraj, Tan-Huy Chu & Je-Jung Lee

  4. Stem Cell & Regenerative Medicine Institute, Samsung Medical Center, Seoul, South Korea

    Minh-Trang Thi Phan, SoonHo Kweon, **ho Kim & Duck Cho

  5. School of Interdisciplinary Bioscience and Bioengineering (I-Bio), POSTECH, Pohang, South Korea

    SoonHo Kweon

  6. Department of Health Sciences and Technology, SAIHST, Sungkyunkwan University, Seoul, South Korea

    **ho Kim & Duck Cho

  7. Bio Research Center, Lugensci Co. Ltd., Bucheon, South Korea

    Jong-Min Lee

  8. Department of Emergency Medicine, Konkuk University Chungju Hospital, Chungju, South Korea

    Ilwoong Hwang

  9. Research Institute of Advanced Materials, Seoul National University, Seoul, South Korea

    Jeehun Park

  10. Department of Materials Science and Engineering, Seoul National University, Seoul, South Korea

    Junsang Doh

  11. Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada

    Seung-Hwan Lee

  12. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, 81 Irwon-dong, Gangnam-gu, Seoul, 06351, Republic of Korea

    Duck Cho

Authors
  1. Jaya Lakshmi Thangaraj
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Contributions

JL Thangaraj, SH Kweon, M-T Phan, MC Vo, and TH Chu performed the research and analyzed data; J Kim, I Hwang, JH Park, J Doh and SH Lee analyzed data; JH Kim, SH Kim, and JM Lee generated genetically engineered K562-OX40L-mb18/-21 cells; JS Doh, D Cho, and JJ Lee designed the research study and JL Thangaraj, SH Kweon, M-T Phan JH Park, JS Doh, GY Song, SY Ahn, SH Jung, HJ Kim, D Cho, and JJ Lee co-wrote the paper.

Corresponding authors

Correspondence to Duck Cho or Je-Jung Lee.

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Thangaraj, J.L., Phan, MT.T., Kweon, S. et al. Expansion of cytotoxic natural killer cells in multiple myeloma patients using K562 cells expressing OX40 ligand and membrane-bound IL-18 and IL-21. Cancer Immunol Immunother 71, 613–625 (2022). https://doi.org/10.1007/s00262-021-02982-9

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