title: A high-throughput assay of NK cell activity in whole blood and its clinical application.
abstract: 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 (51)Cr 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.
authors.affiliation: Department of Microbiology and Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Saet-byul
authors.initials: SB
authors.lastname: Lee
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Junhoe
authors.initials: J
authors.lastname: Cha
authors.affiliation: Department of Surgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Im-kyung
authors.initials: IK
authors.lastname: Kim
authors.affiliation: Department of Microbiology, Ewha Womans University School of Medicine, Seoul, Republic of Korea.
authors.forename: Joo Chun
authors.initials: JC
authors.lastname: Yoon
authors.affiliation: Department of Microbiology and Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Hyo Joon
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Sang Woo
authors.initials: SW
authors.lastname: Park
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Sunjung
authors.initials: S
authors.lastname: Cho
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Dong-ye
authors.initials: DY
authors.lastname: Youn
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Heyja
authors.initials: H
authors.lastname: Lee
authors.affiliation: ATGen Co. Ltd., Sungnam, Republic of Korea.
authors.forename: Choong Hwan
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Department of Microbiology and Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Jae Myun
authors.initials: JM
authors.lastname: Lee
authors.affiliation: Department of Surgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea. Electronic address: kylee117@yuhs.ac.
authors.forename: Kang Young
authors.initials: KY
authors.lastname: Lee
authors.affiliation: Department of Microbiology and Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea. Electronic address: jkim63@yuhs.ac.
authors.forename: Jongsun
authors.initials: J
authors.lastname: Kim
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015179:Colorectal Neoplasms / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007371:Interferon-gamma / Q000097:blood* / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology*; D008297:Male; D008875:Middle Aged
chemical_list: D007371:Interferon-gamma
keywords: ELISA; High-throughput screening; IFN-γ; NK cell activity; Simple assay