Abstract
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-1α 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.
Highlights
Natural killer (NK) cells are a major component of innate immunity and are responsible for immune surveillance
In the negative control samples, more than 99% of K562 cells were stained with carboxyfluorescein succinimidyl ester (CFSE), and spontaneous lysis was less than 2% of them (Figure 1)
NK cytotoxicity of peripheral blood mononuclear cells (PBMCs) was significantly increased with increasing E : T ratios, and the cytotoxicity levels reached 46.5 ± 2.6% at effector to target (E/T) ratio of 32
Summary
Natural killer (NK) cells are a major component of innate immunity and are responsible for immune surveillance They induce direct cytotoxicity or secretion of cytokine/chemokine without recognizing a specific antigen as B and T cells [1]. NK cytotoxicity plays an important role in immune response against infected cells, malignancy, and stressed cells, and involves in pathologic process in various diseases [2,3,4,5]. These cytotoxic functions are markedly variable among individuals, and NK function analysis has become a more routine practice in many diseases [6,7,8].
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