The use of non-radioactive chromium as an alternative to 51Cr in NK assay.
A novel method to measure target cell cytolysis based on the use of 'cold', non-radioactive chromium and on the determination of metal release by graphite furnace atomic absorption spectroscopy (FAAS) is proposed. Natural killer (NK) assays were performed by labelling target cells with chromium as Na2CrO4, and results were compared with those obtained by conventional overnight labelling with 51Cr of targets killed by the same effectors. The cytotoxic capacity of peripheral blood lymphocytes from healthy subjects was evaluated, and NK activity measured with both methods showed a good agreement at each of the tested effector to target cell ratios (between 100:1 and 1:1), with a high and significant coefficient of correlation (r = 0.931, p < 0.0001). The selection of the appropriate Cr concentrations for labelling target cells took into account both the sensitivity of our instrumentation and the possible toxic effects of the metal. A study of the effects of Cr on the cell line (K562) which is usually employed as a target in NK tests showed that Cr could have a detrimental effect on cellular function, with significant numbers of cells with depolarised mitochondria and reduced DNA synthesis after 24 h incubation using Cr levels higher than 15 mumol/l (780 micrograms/l). The method proposed here has a number of advantages, including the use of a non-radioactive tracer, limited costs, high sensitivity and reproducibility, and the possibility of storing samples. In addition, the technique uses a fixed Cr concentration which is known to be non toxic.
Borella
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P
Dipartimento di Scienze Biomediche, Università degli Studi, Modena, Italy.
Bargellini
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A
Salvioli
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S
Medici
C I
CI
Cossarizza
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A
D000328:Adult; D002478:Cells, Cultured; D002857:Chromium; D002860:Chromium Radioisotopes; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D013054:Spectrophotometry, Atomic / Q000379:methods*
D002860:Chromium Radioisotopes; D002857:Chromium