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

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• 作者：Chris Hughes (56)
  Angelica Sette (56)
  Michael Seed (57)
  Fulvio D鈥橝cquisto (56)
  Antonio Manzo (58)
  Tonia L Vincent (59)
  Ngee Han Lim (56)
  Ahuva Nissim (56)
  
  56. Centre for Biochemical Pharmacology ; William Harvey Research Institute ; Barts and The London School of Medicine and Dentistry ; Queen Mary University of London ; London ; EC1M 6BQ ; UK
  57. Medicines Research Group ; School of Health Sport and Bioscience ; University of East London ; Water Lane ; London ; E15 4LZ ; UK
  58. Rheumatology and Translational Immunology Research Laboratories (LaRIT) ; Division of Rheumatology ; IRCCS Policlinico San Matteo Foundation/University of Pavia ; Pavia ; Italy
  59. Kennedy Institute of Rheumatology ; NDORMS ; University of Oxford ; 65 Roosevelt Drive ; Headington ; Oxford ; OX3 7FY ; UK
  
• 刊名：Arthritis Research & Therapy
• 出版年：2014
• 出版时间：August 2014
• 年：2014
• 卷：16
• 期：4
• 全文大小：1,565 KB
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• 刊物主题：Rheumatology; Orthopedics;
• 出版者：BioMed Central
• ISSN：1478-6354

文摘

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.