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 H. Lim, Ahuva Nissim

Research output: Contribution to journalArticle

19 Citations (Scopus)

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
Article numberR151
JournalArthritis Research and Therapy
Volume16
Issue number4
DOIs
Publication statusPublished - Jul 16 2014

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Immunoglobulin Fragments
Interleukin-10
Arthritis
Cartilage
Single-Chain Antibodies
Collagen Type II
Reactive Oxygen Species
Anti-Inflammatory Agents
Therapeutics
Joints
Cytokines
Matrix Metalloproteinase 1
Matrix Metalloproteinases
Knee
Western Blotting
Enzyme-Linked Immunosorbent Assay
Cell Proliferation
Staining and Labeling

ASJC Scopus subject areas

  • Rheumatology
  • Immunology
  • Immunology and Allergy

Cite this

Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency. / Hughes, Chris; Sette, Angelica; Seed, Michael; D'Acquisto, Fulvio; Manzo, Antonio; Vincent, Tonia L.; Lim, Ngee H.; Nissim, Ahuva.

In: Arthritis Research and Therapy, Vol. 16, No. 4, R151, 16.07.2014.

Research output: Contribution to journalArticle

Hughes, Chris ; Sette, Angelica ; Seed, Michael ; D'Acquisto, Fulvio ; Manzo, Antonio ; Vincent, Tonia L. ; Lim, Ngee H. ; Nissim, Ahuva. / Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency. In: Arthritis Research and Therapy. 2014 ; Vol. 16, No. 4.
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AU - Hughes, Chris

AU - Sette, Angelica

AU - Seed, Michael

AU - D'Acquisto, Fulvio

AU - Manzo, Antonio

AU - Vincent, Tonia L.

AU - Lim, Ngee H.

AU - Nissim, Ahuva

PY - 2014/7/16

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N2 - 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.

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