RESUMO
To investigate the initial stages of recognition of the self idiotype (Id) by T cells, we examined the early increase in cytoplasmic free calcium ([Ca2+]i) occurring in murine CD4+ T cells specific for a model Id, Id315, following their interaction with the Id. The changes in [Ca2+]i were monitored with stopped-flow fluorometry by loading T cells with fura 2, a Ca(2+)-binding fluorescent dye. An increase of [Ca2+]i in the Id-specific T cell line was dependent on the presence of both antigen-presenting cells (APC) and Id315. When T cells were mixed with APC pulsed with M315 for 90 min at 37 C, a significant increase in T cell [Ca2+]i was observed within one second. A pronounced elevation in [Ca2+]i was also observed in T cells after their interaction with APC which had been pulsed for 90 min with VL-315 Id-containing proteins (such as VL-315, L315, Fv-315 or Fab'-315 fragments). In contrast, pulsing APC for 5 min with the VL fragment produced little or no change in the [Ca2+]i. These results suggest that VL must be further processed by APC before it can be recognized by T cells. Indeed, a synthetic VL region peptide (positions 91-108, designated as P18) produced an elevation in T cell [Ca2+]i when mixed with APC without pulsing.
