Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency

Chris Hughes, Angelica Sette, Michael Seed, Fulvio D'Acquisto, Antonio Manzo, Tonia L Vincent, Ngee Han Lim, Ahuva Nissim

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Abstract

INTRODUCTION: We previously demonstrated that a single-chain fragment variable (scFv) specific to collagen type II (CII) posttranslationally modified by reactive oxygen species (ROS) can be used to target anti-inflammatory therapeutics specifically to inflamed arthritic joints. The objective of the present study was to demonstrate the superior efficacy of anti-inflammatory cytokines when targeted to inflamed arthritic joints by the anti-ROS modified CII (anti-ROS-CII) scFv in a mouse model of arthritis.

METHODS: Viral interleukin-10 (vIL-10) was fused to anti-ROS-CII scFv (1-11E) with a matrix-metalloproteinase (MMP) cleavable linker to create 1-11E/vIL-10 fusion. Binding of 1-11E/vIL-10 to ROS-CII was determined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and immune-staining of arthritic cartilage, whereas vIL-10 bioactivity was evaluated in vitro by using an MC-9 cell-proliferation assay. Specific in vivo localization and therapeutic efficacy of 1-11E/vIL-10 was tested in the mouse model of antigen-induced arthritis.

RESULTS: 1-11E/vIL-10 bound specifically to ROS-CII and to damaged arthritic cartilage. Interestingly, the in vitro vIL-10 activity in the fusion protein was observed only after cleavage with MMP-1. When systemically administered to arthritic mice, 1-11E/vIL-10 localized specifically to the arthritic knee, with peak accumulation observed after 3 days. Moreover, 1-11E/vIL-10 reduced inflammation significantly quicker than vIL-10 fused to the control anti-hen egg lysozyme scFv (C7/vIL10).

CONCLUSIONS: Targeted delivery of anti-inflammatory cytokines potentiates their anti-arthritic action in a mouse model of arthritis. Our results further support the hypothesis that targeting biotherapeutics to arthritic joints may be extended to include anti-inflammatory cytokines that lack efficacy when administered systemically.

Original languageEnglish
Pages (from-to)R151
JournalArthritis research & therapy
Volume16
Issue number4
DOIs
Publication statusPublished - 16 Jul 2014

Keywords

  • Animals
  • Antibody Specificity
  • Arthritis, Experimental
  • Blotting, Western
  • Cartilage, Articular
  • Collagen Type II
  • Drug Delivery Systems
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunoglobulin Variable Region
  • Immunotherapy
  • Interleukin-10
  • Mice
  • Mice, Inbred C57BL
  • Reactive Oxygen Species
  • Recombinant Fusion Proteins
  • Journal Article
  • Research Support, Non-U.S. Gov't

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