Use of flameless atomic absorption spectroscopy in immune cytolysis for nonradioactive determination of killer cell activity.
We describe here a novel method to evaluate natural killer (NK) cytolytic activity by use of flameless atomic absorption spectroscopy (GF-AAS). This technique may be adopted for use in laboratories equipped with electrothermal atomic absorption spectrometers. Nonradioactive Cr as Na2CrO4 was used to label target cells (K562), and cell lysis was evaluated by measuring Cr released after 4 h of incubation with the effectors. We selected 520 micrograms/L as the optimal dose for labeling targets, between 12 and 20 h as the optimal incubation time, and 10(4) cells as the optimal target size. Advantages of this method include: (a) exclusion of radioactive tracer, with no risk for workers; (b) limited costs; (c) high sensitivity and reproducibility; (d) possibility to store samples; and (e) better control of Cr used for labeling cells due to well-determined, fixed Cr concentrations in the range of nontoxic and linear cellular uptake. Comparison with data obtained by conventional 51Cr labeling of targets killed by the same effectors was excellent, yielding comparable results and corroborating the method.
Borella
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P
Section of Hygiene and Microbiology, Department of Biomedical Sciences, University of Modena, Italy.
Bargellini
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A
Salvioli
S
S
Cossarizza
A
A
D000328:Adult; D002857:Chromium / Q000378:metabolism; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D004915:Leukemia, Erythroblastic, Acute; D012680:Sensitivity and Specificity; D013054:Spectrophotometry, Atomic; D014407:Tumor Cells, Cultured
D002857:Chromium