Regulation of CD40 ligand expression on naive CD4 T cells: a role for TCR but not co-stimulatory signals.

We have investigated the roles of TCR and accessory co-stimulatory signals in the induction of CD40 ligand (CD40L) on CD4 cells. Using naive T cells form TCR transgenic mice, specific for a peptide of pigeon cytochrome c, we show that in contrast to IL-2 secretion, CD40L expression is regulated primarily by signalling through the TCR, is enhanced by accessory molecule interactions, but co-stimulatory signals play little if any role. CD40L was induced at high levels on naive T cells, peaking at 5 h, by class II MHC+ fibroblast antigen-presenting cells (APC) which expressed either ICAM-1, B7-1 or both molecules, whereas only low levels were induced by fibroblasts which did not express any accessory molecules. Differences in intensity and duration of expression were seen following stimulation with ICAM- and B7-expressing APC, with the presence of ICAM resulting in greater and longer expression, although both molecules together were most efficient. The involvement of co-stimulatory signals delivered from accessory molecules was investigated in systems where there was no effect on TCR signalling from adhesive interactions. Anti-CD3, or antigen-pulsed APC lacking accessory molecules, were used to provide the TCR signal, with co-stimulus from either anti-CD28 or accessory molecule-expressing fibroblasts not presenting antigen. Anti-CD3 in the absence of co-stimuli induced high CD40L expression but no IL-2 production and provision of co-stimulatory signals, although inducing large quantities of IL-2, did not increase CD40L expression. In addition, low CD40L expression induced by antigen presented in the absence of accessory molecules was not enhanced by co-stimulation, although IL-2 was strongly up-regulated. These studies suggest that efficient expression of CD40L on naive CD4 cells does require accessory molecules on APC. However, the role of these molecules for CD40L induction, as opposed to IL-2 secretion, is not one of co-stimulation but one of adhesion, presumably allowing stronger or more prolonged signals to be generated through the TCR. The synergistic role of ICAM and B7 during naive CD4 activation was confirmed using dendritic cells as APC, with nearly complete inhibition of CD40L expression as well as IL-2 secretion being seen when both CTLA-4-Ig and anti-LFA-1 were used to block these molecules.