The designer cytokine hyper-interleukin-6 is a potent activator of STAT3-dependent gene transcription in vivo and in vitro.

Interleukin-6 (IL-6) triggers pivotal pathways in vivo. The designer protei n hyper-IL-6 (H-IL-6) fuses the soluble IL-6 receptor (sIL-6R) through an i ntermediate linker with IL-6. The intracellular pathways that are triggered by H-IL-6 are not defined yet. Therefore, we studied the molecular mechani sms leading to H-IL-6-dependent gene activation. H-IL-6 stimulates haptoglo bin mRNA expression in HepG2 cells, which is transcriptionally mediated as assessed by run-off experiments. The increase in haptoglobin gene transcrip tion correlates with higher nuclear translocation of tyrosine-phosphorylate d STAT3 and its DNA binding. As H-IL-6 stimulates STAT3-dependent gene tran scription, we compared the molecular mechanism between IL-6 and H-IL-6. Tra nsfection experiments were performed with STAT3-dependent luciferase constr uct. The same amount of H-IL-6 stimulated luciferase activity faster, stron ger, and for a longer period of time. Dose response experiments showed that a 10-fold lower dose of H-IL-6 stimulated STAT3-dependent gene transcripti on comparable with the higher amount of IL-6. Cotransfection with the gp80 and/or gp130 receptor revealed that the effect of H-IL-6 on STAT3-dependent gene transcription is restricted to the gp80/gp130 receptor ratio. High am ounts of gp130 increased and high amounts of gp80 decreased the effect on H -IL-6-dependent gene tran. scription. To investigate the in vivo effect of H-IL-6 on gene transcription in the liver, H-IL-6 and IL-6 were injected in to C3H mice. H-IL-6 was at least 10-fold more effective in stimulating the DNA binding and nuclear translocation of STAT3, which enhances haptoglobin mRNA and protein expression. Thus H-IL-6 stimulates STAT3-dependent gene tr anscription in liver cells in vitro and in vivo at least 10-fold more effec tively than IL-6. Our results provide evidence that H-IL-6 is a promising d esigner protein for therapeutic intervention during different pathophysiolo gical conditions also in humans.