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

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.

• Authors: Chris Hughes, Angelica Sette, Michael Seed, Fulvio D'Acquisto, Antonio Manzo, Tonia L Vincent, Ngee Han Lim, Ahuva Nissim
• Journal: Arthritis research & therapy (Arthritis Res Ther) Vol. 16 Issue 4 Pg. R151 (Jul 16 2014) ISSN: 1478-6362 [Electronic] England
• PMID: 25029910 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Collagen Type II
• Immunoglobulin Variable Region
• Reactive Oxygen Species
• Recombinant Fusion Proteins
• Interleukin-10

Topics

• Animals
• Antibody Specificity
• Arthritis, Experimental (immunology)
• Blotting, Western
• Cartilage, Articular (immunology)
• Collagen Type II (immunology)
• Drug Delivery Systems (methods)
• Enzyme-Linked Immunosorbent Assay
• Female
• Humans
• Immunoglobulin Variable Region (immunology)
• Immunotherapy (methods)
• Interleukin-10 (administration & dosage)
• Mice
• Mice, Inbred C57BL
• Reactive Oxygen Species (immunology)
• Recombinant Fusion Proteins