Distribution of Several Activating and Inhibitory Receptors on CD3−CD16+ NK Cells and Their Correlation with NK Cell Function in Healthy Individuals

Abstract

The aim of this study was to estimate the distribution and density of a representative set of activating and inhibitory receptors on gated natural killer (NK) cells, as well as on their bright and dim subsets, and to correlate the receptor expression with NK cell activity for healthy individuals on CD3(-)CD16(+) NK cells. We show that in 43 healthy controls NK cell activity against K562 target cells was 37.34% (E:T, 80:1) by standard chromium release assay. The expression of receptors on NK cells and their subsets was analyzed by flow cytometry. The cytotoxic CD3(-)CD16(bright) NK subset constituted 78.97%, while the regulatory CD3(-)CD16(dim) NK subset constituted 21.03% of NK cells. We show the distribution of NKG2D, CD161, CD158a, and CD158b receptors on CD3(-)CD16(+) NK cells in peripheral blood lymphocytes (PBLs), on gated NK cells, and on the CD3(-)CD16(bright) and CD3(-)CD16(dim) subsets. Contrary to CD158a and CD158b killer immunoglobulin-like receptors (KIRs), there is a significant positive correlation of NKG2D and CD161 expression with NK cytotoxicity. We show the kinetics of change in CD3(-)CD16(+)NK/K562 conjugate composition, together with the stronger target binding capacity of CD16(bright) NK cells. Furthermore, we show that after coculture of PBLs with K562 the expression of CD107a, a degranulation marker, on CD3(-)CD16(+)NK cells and subsets is time dependent and significantly higher on the cytotoxic CD3(-)CD16(bright) NK subset. The novel data obtained regarding expression of NK cell activating and inhibitory receptors for healthy individuals may aid in detecting changes that are associated with various diseases.