INFLUENCE OF INTERLEUKIN-6 (IL-6) DIMERIZATION ON FORMATION OF THE HIGH-AFFINITY HEXAMERIC IL-6 RECEPTOR COMPLEX

The high affinity interleukin-6 (IL-6) signaling complex consists of I L-6 and two membrane-associated receptor components: a low affinity bu t specific IL-6 receptor and the affinity converter/signal transducing protein gp130. Monomeric (IL-6(M)) and dimeric (IL-6(D)) forms of Esc herichia coli-derived human IL-6 and the extracellular (''soluble'') p ortions of the IL-6 receptor (sIL-6R) and gp130 have been purified in order to investigate the effect of IL-6 dimerization on binding to the receptor complex. Although IL-6(D) has a higher binding affinity for immobilized sIL-6R, as determined by biosensor analysis employing surf ace plasmon resonance detection, IL-6(M) is more potent than IL-6(D) i n a STAT3 phosphorylation assay. The difference in potency is signific antly less pronounced when measured in the murine 7TD1 hybridoma growt h factor assay and the human hepatoma HepG2 bioassay due to time-depen dent dissociation of 37 degrees C of IL-6 dimers into active monomers. The increased binding affinity of IL-6(D) appears to be due to its ab ility to cross-link two sIL-6R molecules on the biosensor surface. Stu dies of the IL-6 ternary complex formation demonstrated that the reduc ed biological potency of IL-6(D) resulted from a decreased ability of the IL-6(D) .(sIL-6R)(2) complex to couple with the soluble portion of gp130. These data imply that the IL-6-induced dimerization of sIL-6R is not the driving force in promoting formation of the hexameric (IL-6 . IL-6R . gp130)(2) complex. A model is presented whereby the trimeri c complex of IL-6R, pg130, and IL-6(M) forms before the functional hex amer. Due to its increased affinity for the IL-6R but its decreased ab ility to couple with gp130, we suggest that a stable IL-6 dimer may be an efficient IL-6 antagonist.