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A fluorescence NK assay using flow cytometry.

K McGinnes, G Chapman, R Marks

Journal of Immunological Methods

|January 22, 1986

Summary

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A new flow cytometry assay uses carboxyfluorescein diacetate (cFDA) to measure natural killer (NK) cell activity. This method offers a faster, safer alternative to traditional chromium-51 release assays for studying NK cell-mediated cytotoxicity.

Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

Natural killer (NK) cells are crucial for innate immunity, mediating target cell lysis.
Conventional methods for assessing NK cell activity, like the 51Cr-release assay, involve radioisotopes and can be time-consuming.
There is a need for non-radioactive, efficient assays to evaluate NK cell function.

Purpose of the Study:

To develop and validate a novel flow cytometric assay for quantifying NK cell-mediated cytotoxicity.
To compare the performance of the new assay with the established 51Cr-release assay.
To demonstrate the utility of the assay for evaluating various NK cell populations and conditions.

Main Methods:

K562 target cells were labeled with the fluorogenic substrate carboxyfluorescein diacetate (cFDA).

Related Experiment Videos

NK cell activity was assessed by measuring the release of cFDA using flow cytometry.

Results were compared with those obtained from the 51Cr-release assay.

Main Results:

The cFDA-based flow cytometric assay showed comparable results to the 51Cr-release assay.
cFDA labeling was non-toxic to target cells and did not inhibit NK cell-mediated lysis.
The assay successfully evaluated NK-enriched, IFN-alpha-activated, and ALG-inhibited NK cell populations.

Conclusions:

The developed flow cytometric assay provides a reliable, non-radioactive alternative to the 51Cr-release assay for measuring NK cell cytotoxicity.
This assay offers advantages including direct monitoring of target cell lysis, reduced assay time, and avoidance of radioisotope use.
The assay is suitable for diverse applications in NK cell research, including the study of activated and inhibited NK cell populations.