Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display

Marina Kovaleva, Ingo Bussmeyer, Björn Rabe, Joachim Grötzinger, Enge Sudarman, Jutta Eichler, Udo Conrad, Stefan Rose-John, Jürgen Scheller, Marina Kovaleva

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33 Citations (Scopus)

Abstract

Human herpesvirus 8 (HHV-8) encodes several putative oncogenes, which are homologues to cellular host genes known to function in cell cycle regulation, control of apoptosis, and cytokine signaling. Viral interleukin (vIL-6) is believed to play an important role in the pathogenesis of Kaposi's sarcoma as well as primary effusion lymphoma and multicentric Castleman's disease. Therefore, vIL-6 is a promising target for novel therapies directed against HHV-8-associated diseases. By phage display screening of human synthetic antibody libraries, we have selected a specific recombinant antibody, called monoclonal anti-vIL-6 (MAV), binding to vIL-6. The epitope recognized by MAV was localized on the top of the D helix of the vIL-6 protein, which is a part of receptor binding site III. Consequently, MAV specifically inhibits vIL-6-mediated growth of the primary effusion lymphoma-derived cell line BCBL-1 and blocks STAT3 phosphorylation in the human hepatoma cell line HepG2. Since it was previously found that vIL-6 can also induce signals from within the cell, presumably within the endoplasmic reticulum, we fused the recombinant antibody MAV with the endoplasmic retention sequence KDEL (MAV-KDEL). As a result, COS-7 cells expressing MAV-KDEL and synthesizing vIL-6 ceased to secrete the cytokine. Moreover, we observed that vIL-6 that was bound to MAV-KDEL and retained in the endoplasmic reticulum did not induce STAT3 phosphorylation in HepG2 cells. We conclude that the activity of the intracellularly retained vIL-6 protein is neutralized by MAV-KDEL. Our results might represent a novel therapeutic strategy to neutralize virally encoded growth factors or oncogenes.
Original languageEnglish
Pages (from-to)8510-20
Number of pages11
JournalJournal of Virology
Volume80
Issue number17
DOIs
Publication statusPublished - Sep 2006

Keywords

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antibody Specificity
  • COS Cells
  • Cell Line
  • Cercopithecus aethiops
  • Endoplasmic Reticulum
  • Escherichia coli
  • Humans
  • Immunoglobulin Variable Region
  • Interleukin-6
  • Neutralization Tests
  • Peptide Library
  • Recombinant Proteins
  • Signal Transduction
  • Viral Proteins

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    Kovaleva, M., Bussmeyer, I., Rabe, B., Grötzinger, J., Sudarman, E., Eichler, J., Conrad, U., Rose-John, S., Scheller, J., & Kovaleva, M. (2006). Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display. Journal of Virology, 80(17), 8510-20. https://doi.org/10.1128/JVI.00420-06