Cell mediated lympholysis; a modified technique using 111indium-oxine-labelled targets

Abstract

The isotope 51Cr generally used in the cell mediated lympholysis (CML) assay suffers from the disadvantage of low specific activity, poor incorporation and high spontaneous release, limiting the CML assay to 4–6 h. We have labelled PHA derived human lymphoblasts with the isotope 111indium (using 111indium-oxine) and evaluated these cells as targets in CML. The level of 111In-oxine incorporation decreased rapidly in the presence of serum; in the absence of serum approximately 85% of the available isotope in the supernatant was incorporated into the blasts. Under the labelling conditions used, spontaneous release was 1.6–2%/h on average allowing an effector phase of 18 h. About 5–8% of the released isotope was reutilized by the effector cells during an 18 h incubation period. Extending the CML assay from 6 to 18 h greatly increased the cytotoxicity. At an effector to target ratio of 25:1, the average per cent specific release increased from 15 to 50%. The use of 111In-oxine labelled targets in the CML therefore increases the sensitivity of the test and allows fewer effector and target cells to be used as compared with 51Cr techniques.