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bioRxiv · 10.64898/2026.03.02.708954

Mapping the MIF-2 Chemokine Interactome Reveals MIF-2-CCL20 Complex Formation in Liver Fibrosis

Hoffmann, A.·Brandhofer, M.·He, Y.·Bushati, G.·Siminkovitch, E.·Xavier, B.·Otabil, M. K.·Zhang, L.·Ebert, S.·Holzner, M.·Krammer, C.·Hille, K.·Wichmann, C.·Niess, H.·El-Bounkari, O.·Christian, W.·Kapurniotu, A.·von Hundelshausen, P.·Scheiermann, P.·Bernhagen, J.
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Abstract

D-dopachrome tautomerase (D-DT/MIF-2) is an inflammatory cytokine, atypical chemokine (ACK) and member of the macrophage migration-inhibitory factor (MIF) family. While interactions among classical chemokines (CKs) are established, ACK-CK interactions remain underexplored. Here, we screened for MIF-2 binding-partners using a protein array encompassing all CKs and selected ACKs, and validated candidate interactors by surface-plasmon resonance. CCL20/MIP-3 was prioritized based on RNA-sequencing suggesting induction during liver fibrosis. MIF-2/CCL20 complex formation was verified by microscale thermophoresis and interaction interfaces mapped using peptide array and in-silico modeling. The deduced binding-site near the MIF-2 tautomerase pocket was consistent with inhibition of its tautomerase activity by CCL20. We found both proteins abundantly expressed in human liver tissue, with a positive correlation. Pull-down confirmed complex formation and proximity ligation assay demonstrated MIF-2/CCL20 complexes in liver in situ, with higher levels in fibrotic tissue. Functionally, MIF-2/CCL20 complexes suppressed MIF-2-driven CD4 T-cell chemotaxis and fibroblast IL-6 secretion, indicating modulation of immune and stromal responses. This study extends the ACK interactome to MIF-2 and suggests ACK/CK complexes modulate chemokine activities in liver fibrosis.

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Hoffmann, A., Brandhofer, M., He, Y., Bushati, G., Siminkovitch, E., Xavier, B., Otabil, M. K., Zhang, L., Ebert, S., Holzner, M., Krammer, C., Hille, K., Wichmann, C., Niess, H., El-Bounkari, O., Christian, W., Kapurniotu, A., von Hundelshausen, P., Scheiermann, P., Bernhagen, J.. 2026-03-03. Mapping the MIF-2 Chemokine Interactome Reveals MIF-2-CCL20 Complex Formation in Liver Fibrosis. https://doi.org/10.64898/2026.03.02.708954

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