Cooperative effect of interferon-gamma and tumor necrosis factor-alpha on the induction of the class II antigen-associated invariant chain expression.

The regulation of the invariant chain (Ii) expression was studied in the human colon carcinoma cell line HT-29 that constitutively expressed neither Ii nor class II antigens. Upon stimulation of HT-29 cells with a combination of human recombinant tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma), expression of mRNA and protein of the invariant chain were induced. In contrast, administration of TNF-alpha or IFN-gamma alone had no effect. A delayed induction of Ii mRNA, which was first detected 10-12 h after stimulation, was observed; this suggests an indirect regulatory mechanism. Stimulation with both IFN-gamma and TNF-alpha led to the co-expression of class II antigens with the invariant chain. In order to study the genetic basis for this stimulation the murine invariant chain gene (800 bp 5' flanking sequences and the structural gene) was transfected into HT-29 cells and transfected cells were tested for the ability to respond to IFN-gamma and TNF-alpha. Simultaneous application of both cytokines had a strong effect on the induction of the murine invariant chain. IFN-gamma alone had no effect and TNF-alpha only marginally stimulates murine invariant chain expression. The transfection experiment indicates that the murine invariant chain gene construct contains the structural elements which are responsible for regulation with IFN-gamma and TNF-alpha. We determined whether the cooperative effect of TNF-alpha and IFN-gamma is also found in vivo. Stimulations of mice were performed with TNF-alpha, IFN-gamma and a combination of both. The immunohistological analysis of kidney tissue sections revealed that TNF-alpha had no effect on Ii and Ia expression. Upon IFN-gamma treatment a minor subset of renal tubules showed staining for Ii, and less prominently also for Ia. However, simultaneous application of both cytokines led a strong induction of both Ii and Ia antigens in renal epithelial cells, thus suggesting that this synergistic effect potentially occurs under physiological conditions.