A modified assay for measuring thymocyte co-stimulatory activity

Abstract

The observation that murine thymocytes increase their proliferation to interleukin 1 (IL-1) in the presence of phytohemagglutinin (PHA) when pre-incubated with interleukin 2 (IL-2) allowed the introduction of a modified assay for the measurement of IL-1 or the search of thymocyte-inducing proliferative activities in biological samples. Pre-incubation of thymocytes for 24 hr with 50 u/ml IL-2, followed by washings, elicited their maximal response to IL-1 in the usual lymphocyte activating factor (LAF) assay. This suggests that sequential events lead to thymocyte activation. The responsiveness is three to five fold greater than, and the total time of assay is the same as that of the LAF assay. Interestingly, pre-incubation with IL-2 renders thymocytes more sensitive than responsive to crude monocyte conditioned media. The use of the MTT colorimetric method for the assessment of thymocyte proliferation, and of the lectin jacalin as a co-mitogen are suggested as alternatives to be used in co-stimulatory assays.

Effects of plant lectins on in vitro fibroblast proliferation

“…Rapidness, precision, lacks of any radioisotope (Mosmann, 1983) and same or higher sensitivity than 3 H -timidine incorporation assay (Dalmau et al, 1993;Gieni et al, 1995) are the advantages of the method. In the original method, the colored product was read at 570nm (Mosmann, 1983) however we used 490nm.…”