Evaluation of NK cell function by flowcytometric measurement and impedance based assay using real-time cell electronic sensing system.
Although real-time cell electronic sensing (RT-CES) system-based natural killer (NK) cytotoxicity has been introduced, it has not been evaluated using human blood samples. In present study, we measured flowcytometry based assay (FCA) and RT-CES based NK cytotoxicity and analyzed degranulation activity (CD107a) and cytokine production. In 98 healthy individuals, FCA with peripheral blood mononuclear cells (PBMCs) at effector to target (E/T) ratio of 32 revealed 46.5 ± 2.6% cytolysis of K562 cells, and 23.5 ± 1.1% of NK cells showed increased degranulation. In RT-CES system, adherent NIH3T3 target cells were resistant to basal killing by PBMC or NK cells. NK cell activation by adding IL-2 demonstrated real-time dynamic killing activity, and lymphokine-activated PBMC (E/T ratio of 32) from 15 individuals showed 59.1 ± 6.2% cytotoxicity results after 4 hours incubation in RT-CES system. However, there was no significant correlation between FCA and RT-CES cytotoxicity. After K562 target cell stimulation, PBMC produced profound proinflammatory and immunoregulatory cytokines/chemokines including IL-2, IL-8, IL-10, MIP-1 α β , IFN- γ , and TNF- α , and cytokine/chemokine secretion was related to flowcytometry-based NK cytotoxicity. These data suggest that RT-CES and FCA differ in sensitivity, applicability and providing information, and further investigations are necessary in variable clinical conditions.
Department of Biomedical Science, Graduate School, Catholic University of Korea, 505 Banpo-Dong, Seocho-Ku, Seoul 137-701, Republic of Korea.
Ki-Hyun
KH
Park
Hyesun
H
Park
Myungshin
M
Kim
Yonggoo
Y
Kim
Kyungja
K
Han
Eun-Jee
EJ
Oh
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D015550:Cell Degranulation; D018925:Chemokines / Q000096:biosynthesis; D003199:Computer Systems; D003602:Cytotoxicity, Immunologic; D017097:Electric Impedance; D004581:Electronics / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology / Q000502:physiology; D052118:Lysosomal-Associated Membrane Protein 1 / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells
D018925:Chemokines; D052118:Lysosomal-Associated Membrane Protein 1