Amended recombinant cells (ARCs): an economical and surprisingly effective production and delivery vehicle for recombinant bovine IFN-gamma

J Control Release. 2005 Oct 3;107(2):189-202. doi: 10.1016/j.jconrel.2005.06.008.

Abstract

Recombinant Pseudomonas fluorescens cells, expressing over 40% protein as bovine interferon-gamma (IFN-gamma), were chemically fixed to sterilize the culture and amend the bacterial cell wall. When killed and fixed recombinant cells, termed here amended-recombinant-cells (ARCs), were assayed for interferon activity, we obtained the following surprising results: 1) sterilization and fixation did not inactivate ARC-encapsulated IFN-gamma; 2) ARC-encapsulated IFN-gamma and soluble, recombinant IFN-gamma were equally active in vitro but proteolysis was required for release of the ARC cytokine; and 3) ARC-encapsulated IFN-gamma was active in vivo with optimal adjuvant activity at a dose about 1000-fold less than previously reported for soluble, recombinant IFN-gamma and 100-fold less than doses which induced adverse systemic effects. The mechanism by which ARC-encapsulation increased IFN-gamma activity in vivo remains uncertain. However, our in vitro results show that sustained release of soluble IFN-gamma is a likely factor. The ARC production and delivery system achieves enhanced adjuvant activity with reduced risk of systemic effects, and the low cost of IFN-gamma production offers new opportunities for the use of this important cytokine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / administration & dosage*
  • Blotting, Western
  • Cattle
  • Cell Wall / chemistry
  • Clinical Trials as Topic
  • Cloning, Molecular
  • Cytokines / administration & dosage
  • Drug Delivery Systems* / economics
  • Drug Delivery Systems* / veterinary
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Gene Expression / drug effects
  • Genes, MHC Class II
  • Haptoglobins / biosynthesis
  • Interferon-gamma / administration & dosage*
  • Interferon-gamma / biosynthesis*
  • Male
  • Pseudomonas aeruginosa / metabolism*
  • Recombinant Proteins
  • Sterilization
  • Vesicular stomatitis Indiana virus / drug effects
  • Viral Plaque Assay

Substances

  • Antiviral Agents
  • Cytokines
  • Haptoglobins
  • Recombinant Proteins
  • Interferon-gamma