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Oxygen‐Reactive Metabolites Are Not Detected at the Effector‐Target Interface During Natural Killing
Author(s) -
Storkus Walter J.,
Dawson Jeffrey R.
Publication year - 1986
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.39.5.547
Subject(s) - luminol , chemiluminescence , effector , cytolysis , natural killer cell , biology , reactive oxygen species , biochemistry , cytotoxicity , chemistry , in vitro , chromatography
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.