Oxygen-reactive metabolites are not detected at the effector-target interface during natural killing.
Oxygen-reactive metabolites are not detected at the natural killer (NK)-target cell interface in quantities comparable to those seen for other effector-target cell interactions. A novel luminol-coated target cell chemiluminescence assay is described in which luminol is conjugated to the target cell surface with the bifunctional crosslinker 3,3'-dithiobis(propionic acid N-hydroxysuccinimide ester) (DSP). This modification of the basic chemiluminescence assay precludes exclusion of the detection system from the tightly occluded intercellular junction, a possible deficiency in previous investigations. Luminol conjugation does not affect NK-mediated conjugate formation or cytolysis. As NK activity is enriched by a standard series of effector fractionation procedures, chemiluminescence generated against labeled target cells diminishes. Residual chemiluminescence in the most highly NK-active effector fraction is ablated upon antibody and complement depletion of MO2+ cells. This indicates that monocyte contamination is the source of luminol-detectable oxygen-reactive metabolites.
Storkus
W J
WJ
Dawson
J R
JR
D002469:Cell Separation; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007113:Immunity, Innate; D007694:Killer Cells, Natural / Q000502:physiology*; D008163:Luminescent Measurements; D008165:Luminol; D009000:Monocytes / Q000502:physiology; D010100:Oxygen / Q000502:physiology*
D008165:Luminol; D010100:Oxygen