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A high-throughput assay of NK cell activity in whole blood and its clinical application

 Saet-byul Lee  ;  Junhoe Cha  ;  Im-kyung Kim  ;  Joo Chun Yoon  ;  Hyo Joon Lee  ;  Sang Woo Park  ;  Sunjung Cho  ;  Dong-Ye Youn  ;  Heyja Lee  ;  Choong Hwan Lee  ;  Jae Myun Lee  ;  Kang Young Lee 
 Biochemical and Biophysical Research Communications, Vol.445(3) : 584-590, 2014 
Journal Title
 Biochemical and Biophysical Research Communications 
Issue Date
Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms/blood* ; Colorectal Neoplasms/immunology ; Enzyme-Linked Immunosorbent Assay/methods ; Female ; High-Throughput Screening Assays/methods* ; Humans ; Interferon-gamma/blood* ; Interferon-gamma/immunology ; Killer Cells, Natural/immunology* ; Male ; Middle Aged
ELISA ; High-throughput screening ; IFN-γ ; NK cell activity ; Simple assay
Natural killer (NK) cells are lymphocytes of the innate immune system and have the ability to kill tumor cells and virus-infected cells without prior sensitization. Malignant tumors and viruses have developed, however, strategies to suppress NK cells to escape from their responses. Thus, the evaluation of NK cell activity (NKA) could be invaluable to estimate the status and the outcome of cancers, viral infections, and immune-mediated diseases. Established methods that measure NKA, such as 51Cr release assay and CD107a degranulation assay, may be used to determine NK cell function, but they are complicated and time-consuming because they require isolation of peripheral blood mononuclear cells (PBMC) or NK cells. In some cases these assays require hazardous material such as radioactive isotopes. To overcome these difficulties, we developed a simple assay that uses whole blood instead of PBMC or isolated NK cells. This novel assay is suitable for high-throughput screening and the monitoring of diseases, because it employs serum of ex vivo stimulated whole blood to detect interferon (IFN)-γ secreted from NK cells as an indicator of NKA. After the stimulation of NK cells, the determination of IFNγ concentration in serum samples by enzyme-linked immunosorbent assay (ELISA) provided a swift, uncomplicated, and high-throughput assay of NKA ex vivo. The NKA results microsatellite stable (MSS) colorectal cancer patients was showed significantly lower NKA, 263.6 ± 54.5 pg/mL compared with healthy subjects, 867.5 ± 50.2 pg/mL (p value <0.0001). Therefore, the NKA could be utilized as a supportive diagnostic marker for microsatellite stable (MSS) colorectal cancer.
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1. College of Medicine (의과대학) > Dept. of Surgery (외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Im Kyung(김임경) ORCID logo https://orcid.org/0000-0001-8505-5307
Kim, Jong Sun(김종선) ORCID logo https://orcid.org/0000-0002-3149-669X
Lee, Kang Young(이강영)
Lee, Saet Byul(이샛별)
Lee, Jae Myun(이재면) ORCID logo https://orcid.org/0000-0002-5273-3113
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