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...

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Bibliographic Details
Published in:Journal of leukocyte biology 1986-05, Vol.39 (5), p.547-557
Main Authors: Storkus, Walter J., Dawson, Jeffrey R.
Format: Article
Language:English
Subjects:
3,3′‐dithiobis(propionic acid N‐hydroxysuccinimide ester)
Biological and medical sciences
Cell Separation
Cytotoxic reactions (adcc reaction, cell-mediated lympholysis, complement-dependent cytotoxicity and others)
Cytotoxicity Tests, Immunologic
Cytotoxicity, Immunologic
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Humans
Immunity, Innate
Immunobiology
Immunological reactions in vitro
Killer Cells, Natural - physiology
Luminescent Measurements
Luminol
Monocytes - physiology
natural killer
Oxygen - physiology
oxygen intermediates
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Summary: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.
ISSN:0741-5400
1938-3673
DOI:10.1002/jlb.39.5.547