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

Abstract

IntroductionWe 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.MethodsViral 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.Results1-11E/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).ConclusionsTargeted 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.

Immunocytokines: a novel class of products for the treatment of chronic inflammation and autoimmune conditions

“…The resulting fusion protein was able to inhibit disease progression in a mouse model of antigen-induced arthritis. Similar results were obtained by combining the antibody to the soluble portion of mTNFRII receptor fused to an Fc moiety [52,69].…”

IL-10 reduces apoptosis and extracellular matrix degradation after injurious compression of mature articular cartilage

“…The major difficulty seems to be the administration of IL-10 and calculation of its effective dosage in vivo. Targeted gene therapy might offer an effective approach, which allows a disease-regulated overexpression of IL-10 within the damaged cartilage joint 44,45 . Another promising strategy might be the administration of IL-10 in in vitro tissue engineering protocols since Tsuchida et al revealed that expanded chondocytes have impaired ability to produce IL-10 in vitro 30 .…”

Targeting Extracellular Vesicles to the Arthritic Joint Using a Damaged Cartilage-Specific Antibody

“…We have produced and characterized an antibody that is specific to CII post-translationally modified by oxidants (anti-ROS-CII). This unique antibody has been validated in a series of studies demonstrating strong immunostaining on arthritic but not healthy cartilage, specific targeting to arthritic joint in vivo, and attenuation of knee inflammation by drug targeted by anti-ROS-CII (28,31). Once grafted onto EV, the anti-ROS-CII antibody acted as an arthritic cartilage-specific navigator, leading to an accumulation of EV selectively to arthritic joints with a peak at 18 h post-i.v.…”