title: Targeting of viral interleukin-10 with an antibody fragment specific to damaged arthritic cartilage improves its therapeutic potency.
abstract: BACKGROUND
  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.forename: Chris
authors.initials: C
authors.lastname: Hughes
authors.forename: Angelica
authors.initials: A
authors.lastname: Sette
authors.forename: Michael
authors.initials: M
authors.lastname: Seed
authors.forename: Fulvio
authors.initials: F
authors.lastname: D'Acquisto
authors.forename: Antonio
authors.initials: A
authors.lastname: Manzo
authors.forename: Tonia L
authors.initials: TL
authors.lastname: Vincent
authors.forename: Ngee Han
authors.initials: NH
authors.lastname: Lim
authors.forename: Ahuva
authors.initials: A
authors.lastname: Nissim
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D001169:Arthritis, Experimental / Q000276:immunology*; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000276:immunology; D024043:Collagen Type II / Q000276:immunology*; D016503:Drug Delivery Systems / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007167:Immunotherapy / Q000379:methods*; D016753:Interleukin-10 / Q000008:administration & dosage*; D051379:Mice; D008810:Mice, Inbred C57BL; D017382:Reactive Oxygen Species / Q000276:immunology; D011993:Recombinant Fusion Proteins
chemical_list: D024043:Collagen Type II; D007135:Immunoglobulin Variable Region; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10