Investigation of early T cell activation: Analysis of the effect of specific antigen, interleukin 2 and monoclonal antibodies on intracellular free calcium concentration

Abstract

The three mitogenic anti-T3 antibodies, UCHT1, anti-Leu-4 and WT-32, all produce a rapid increase in T cell intracellular Ca²⁺ ([Ca²⁺]j) in all individuals, as measured by quin2 tetra-acetoxymethyl ester fluorescence. This indicates that the lack of responsiveness of approximately 30% of individuals to UCHT1 in proliferation assays is not due to failure of the antibody to elicit Ca²⁺ mobilization and that a rise in [Ca²⁺]j is per se not adequate to induce cell division. Another mitogenic antibody, WT-31, which is directed against the constant portion of the T cell receptor, did not, however, produce a rapid calcium rise in peripheral blood T cells. The clone HA1.7 gave a similar Ca²⁺response to UCHT1. WT-31 did not induce a rise in [Ca²⁺]j, nor did the specific antigen to which the clone responded. Accessory cells may be required to induce Ca²⁺mobilization with these ligands. There was no response to IL2, or an antibody (anti- Tac) to the IL2 receptor. In contrast to peripheral blood T cells treatment of HA1.7 with WT-31 led to an enhancement of the calcium response to subsequent UCHT1 addition. Furthermore, cross-linking of WT-31 on the surface of HA1.7 cells did produce a small rise in [Ca²⁺]j. The IL2-independent malignant T cell line, HUT78, exhibited a calcium response to both UCHT1 and WT-32. Both of these responses occurred without cross-linking. The T cell receptor is closely associated with cell-surface proteins, including the T3 antigen and these studies confirm the importance of the T3 antigen in T cell activa- tion. They also suggest that the relationship between the T cell receptor and the T3 antigen may vary in T cells in different proliferative states.