Original Article

Korean J Hematol 2006; 41(1):

Published online March 30, 2006

https://doi.org/10.5045/kjh.2006.41.1.8

© The Korean Society of Hematology

K562 세포주와 IL-2를 이용하여 말초혈액 단핵구로부터 자연살세포의 선택적 증폭

조 덕, 신시원, 박정선, 강현규, 김상기, Nguyen Pham TN, Zhu XW, 신명근, 서순팔, 양동욱, 남종희, 김영진, 이제중

전남대학교 의과대학 화순전남대학교병원 암백신팀, 진단검사의학교실

Selective Expansion of Natural Killer Cells from Peripheral Blood Mononuclear Cells by K562 Cell Line and IL-2

Duck Cho, Shi Won Shin, Jung Sun Park, Hyun Kyu Kang, Sang Ki Kim, Than Nhan Nguyen Pham, Xiao Wei Zhu, Myung Geun Shin, Soon Pal Suh, Dong Wook Ryang, Jong Hee Nam, Young Jin Kim, Je Jung Lee

Cancer Vaccine Team, Chonnam National University Hwasun Hospital, Jeonnam, Department of Laboratory
Medicine, Chonnam National University Medical School, Gwangju, Korea

Abstract

Background:
Several attempts have been made to expand human NK cells from peripheral blood mononuclear cells (PBMCs). This study examined the selective expansion of NK cells using interleukin 2 (IL-2) plus the K562 cell line, the expression of the NK cell receptors, and the cytotoxic activity.
Methods:
The PBMCs from seven healthy volunteers were cultured in a medium containing the IL-2 plus the K562 cell line for 14 days. The expression of the activating and inhibitory receptors on the resting NK cells and the 72 hr-expanded NK cells were analyzed. A flow cytometric cytotoxic assay was used to determined the killing activity of the non-expanded NK cells and the 7 day-expanded NK cells against the K562 target cells.
Results:
The NK cells from PBMCs expanded 4.5-fold after 7 days, and contained 56.5% CD3-CD56+ cells. The IL-2 or IL-2 plus K562 increased the expression levels of CD158b (MFI, mean florescence intensity), CD158e1/e2 (MFI), and NKp44 (MFI), while it decreased the expression levels of NKp30 (%), CD16 (MFI), and 2B4 (MFI). The non-expanded NK cells lysed 9.0% and 27.6% of the K562 target cells in the 1:1 and 5:1 effector and target ratio, respectively, and the 7-day expanded NK cells lysed 36.9% and 57.2% of the K562 target cells, respectively.
Conclusion:
The selective expansion of CD3-CD56+ NK cells occurred only during 7 days of culture. IL-2 or IL-2 plus the K562 cells altered the expression of various activating and inhibitory receptors of NK cells, and the cytotoxicity of the expanded NK cells was higher than in the non-expanded cells.

Keywords Natural killer cells, K562, IL-2, Expansion

Article

Original Article

Korean J Hematol 2006; 41(1): 8-15

Published online March 30, 2006 https://doi.org/10.5045/kjh.2006.41.1.8

Copyright © The Korean Society of Hematology.

K562 세포주와 IL-2를 이용하여 말초혈액 단핵구로부터 자연살세포의 선택적 증폭

조 덕, 신시원, 박정선, 강현규, 김상기, Nguyen Pham TN, Zhu XW, 신명근, 서순팔, 양동욱, 남종희, 김영진, 이제중

전남대학교 의과대학 화순전남대학교병원 암백신팀, 진단검사의학교실

Selective Expansion of Natural Killer Cells from Peripheral Blood Mononuclear Cells by K562 Cell Line and IL-2

Duck Cho, Shi Won Shin, Jung Sun Park, Hyun Kyu Kang, Sang Ki Kim, Than Nhan Nguyen Pham, Xiao Wei Zhu, Myung Geun Shin, Soon Pal Suh, Dong Wook Ryang, Jong Hee Nam, Young Jin Kim, Je Jung Lee

Cancer Vaccine Team, Chonnam National University Hwasun Hospital, Jeonnam, Department of Laboratory
Medicine, Chonnam National University Medical School, Gwangju, Korea

Abstract

Background:
Several attempts have been made to expand human NK cells from peripheral blood mononuclear cells (PBMCs). This study examined the selective expansion of NK cells using interleukin 2 (IL-2) plus the K562 cell line, the expression of the NK cell receptors, and the cytotoxic activity.
Methods:
The PBMCs from seven healthy volunteers were cultured in a medium containing the IL-2 plus the K562 cell line for 14 days. The expression of the activating and inhibitory receptors on the resting NK cells and the 72 hr-expanded NK cells were analyzed. A flow cytometric cytotoxic assay was used to determined the killing activity of the non-expanded NK cells and the 7 day-expanded NK cells against the K562 target cells.
Results:
The NK cells from PBMCs expanded 4.5-fold after 7 days, and contained 56.5% CD3-CD56+ cells. The IL-2 or IL-2 plus K562 increased the expression levels of CD158b (MFI, mean florescence intensity), CD158e1/e2 (MFI), and NKp44 (MFI), while it decreased the expression levels of NKp30 (%), CD16 (MFI), and 2B4 (MFI). The non-expanded NK cells lysed 9.0% and 27.6% of the K562 target cells in the 1:1 and 5:1 effector and target ratio, respectively, and the 7-day expanded NK cells lysed 36.9% and 57.2% of the K562 target cells, respectively.
Conclusion:
The selective expansion of CD3-CD56+ NK cells occurred only during 7 days of culture. IL-2 or IL-2 plus the K562 cells altered the expression of various activating and inhibitory receptors of NK cells, and the cytotoxicity of the expanded NK cells was higher than in the non-expanded cells.

Keywords: Natural killer cells, K562, IL-2, Expansion

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