HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

Tumor-targeting properties of novel immunocytokines based on murine IL1β and IL6.

Abstract
There is an increasing biotechnological interest in 'arming' therapeutic antibodies with bioactive payloads. Many antibody-cytokine fusion proteins (immunocytokines) have been described and some of these biopharmaceuticals have progressed to clinical studies. Here, we describe for the first time the expression and in vivo characterization of immunocytokines based on murine IL1β and IL6. These potent pro-inflammatory cytokines were fused at the N-terminus or at the C-terminus of the monoclonal antibodies F8 (specific to the alternatively-spliced extra-domain A domain of fibronectin, a marker of tumor angiogenesis). All immunocytokines retained the binding properties of the parental antibody and were homogenous, when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography, except for the N-terminal fusion of IL1β which revealed the presence of glycosylated species. When analyzed by quantitative biodistribution analysis using radioiodinated protein preparations, F8 fusions with IL6 revealed a preferential accumulation at the tumor site for both cytokine orientations, whereas IL1β fusions exhibited lower tumor to organ ratios and a slower blood clearance profile. The fusion proteins with the cytokine payload at the C-terminus were studied in therapy experiments in immunocompetent mice bearing F9 tumors. Immunocytokines based on IL1β resulted in 10% body weight loss at a 5-µg dose, whereas IL6-based products caused a 5% body weight loss at a 225-µg dose. Both F8-IL1β and F8-IL6 exhibited a <50% inhibition of tumor growth rate, which was substantially lower than the one previously reported for F8-TNF, a closely related pro-inflammatory immunocytokine. This study indicates that IL6 can be efficiently delivered to the tumor neo-vasculature by fusion with the F8 antibody. While F8-IL6 was not as potent as other F8-based immunocytokines that exhibit similar biodistribution profiles, the fusion protein sheds light on the different roles of pro-inflammatory cytokines in boosting immunity against the tumor.
AuthorsChristian Hess, Dario Neri
JournalProtein engineering, design & selection : PEDS (Protein Eng Des Sel) Vol. 27 Issue 6 Pg. 207-13 (Jun 2014) ISSN: 1741-0134 [Electronic] England
PMID24795343 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Copyright© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: [email protected].
Chemical References
  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • F8 monoclonal antibody
  • Fibronectins
  • IL1B protein, mouse
  • Interleukin-1beta
  • Interleukin-6
  • Recombinant Fusion Proteins
Topics
  • Animals
  • Antibodies, Monoclonal (administration & dosage, biosynthesis, pharmacokinetics)
  • Antibodies, Monoclonal, Humanized
  • CHO Cells
  • Cell Line, Tumor
  • Cricetinae
  • Cricetulus
  • Drug Delivery Systems
  • Drug Screening Assays, Antitumor
  • Female
  • Fibronectins (immunology)
  • Humans
  • Interleukin-1beta (administration & dosage, biosynthesis, pharmacokinetics)
  • Interleukin-6 (administration & dosage, biosynthesis, pharmacokinetics)
  • Mice
  • Mice, 129 Strain
  • Neoplasm Transplantation
  • Protein Binding
  • Recombinant Fusion Proteins (administration & dosage, biosynthesis, pharmacokinetics)
  • Teratocarcinoma (drug therapy, pathology)
  • Tissue Distribution
  • Tumor Burden (drug effects)

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: