The effect of interleukin-10 knock-out and overexpression on neointima formation in hypercholesterolemic APOE*3-Leiden mice

Eefting, Daniel; Schepers, Abbey; De Vries, Margreet R.; Pires, Nuno M.M.; Grimbergen, Jos M.; Lagerweij, Tonny; Nagelkerken, Lex M.; Monraats, Pascalle S.; Jukema, J. Wouter; van Bockel, J. Hajo; Quax, Paul H.A.

doi:10.1016/j.atherosclerosis.2006.09.032

« Previous Next »Atherosclerosis
Volume 193, Issue 2 , Pages 335-342, August 2007

The effect of interleukin-10 knock-out and overexpression on neointima formation in hypercholesterolemic APOE*3-Leiden mice

Daniel Eefting
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
- Department of Vascular Surgery, Leiden University Medical Center, Leiden, The Netherlands
- These authors contributed equally to this work.
Abbey Schepers
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
- Department of Vascular Surgery, Leiden University Medical Center, Leiden, The Netherlands
- These authors contributed equally to this work.
Margreet R. De Vries
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
Nuno M.M. Pires
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
- Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands
Jos M. Grimbergen
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
Tonny Lagerweij
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
Lex M. Nagelkerken
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
Pascalle S. Monraats
- Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands
J. Wouter Jukema
- Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands
J. Hajo van Bockel
- Department of Vascular Surgery, Leiden University Medical Center, Leiden, The Netherlands
Paul H.A. Quax
- Gaubius Laboratory, TNO-Quality of Life, Leiden, The Netherlands
- Department of Vascular Surgery, Leiden University Medical Center, Leiden, The Netherlands
- Corresponding author at: Gaubius Laboratory, TNO-Quality of Life, Department of Biomedical Research, Leiden University Medical Center, Zernikedreef 9, P.O. Box 2215, 2301 CE Leiden, The Netherlands. Tel.: +31 715181509; fax: +31 715181904.

Received 12 May 2006; received in revised form 22 September 2006; accepted 27 September 2006. published online 08 November 2006.

Abstract

Objective

Inflammatory factors are thought to play a regulatory role in restenosis. Interleukin-10 (IL10) is an important anti-inflammatory cytokine with anti-atherogenic potentials. The aim of this study was to assess the effects of IL10 modulation on cuff-induced neointima formation in hypercholesterolemic APOE*3-Leiden mice.

Methods

The involvement of IL10 in neointima formation was studied in a hypercholesterolemic mouse model of cuff-induced stenosis of the femoral artery by IL10 knocking-out or overexpression procedures. IL10^+/− mice were crossbred with APOE*3-Leiden mice to generate hypercholesterolemic APOE*3-LeidenIL10^−/− mice. To achieve IL10 overexpression in APOE*3-Leiden mice, a single intramuscular injection of a murine IL10 overexpression plasmid was performed followed by electroporation.

Results

Knocking-out IL10, in hypercholesterolemic APOE*3-Leiden mice, resulted in a significant 1.9-fold increase of neointima surface as compared to APOE*3-LeidenIL10^+/+ littermates (p=0.02). Conversely, a marked 45% inhibition on cuff-induced neointima formation was obtained after IL10 overexpression (p=0.02). Electrodelivery of IL10 vector leads to detectable IL10 serum levels, with a sustained expression over the experimental period of 3 weeks. IL10 overexpression reduced plasma cholesterol levels in APOE*3-Leiden mice, whereas IL10 deficiency in these mice did not lead to altered cholesterol levels as compared to the IL10^+/+ group. Finally, IL10 overexpression stimulated endogenous IL10 mRNA expression in the spleen and reduced the transcriptional responses of several pro-inflammatory cytokines.

Conclusion

Here, we clearly demonstrate the role of IL10 in the development of neointima formation in hypercholesterolemic mice and the potential therapeutic effect of non-viral electrodelivery of IL10 cDNA to inhibit post-angioplasty restenosis.

Keywords: Restenosis, Cytokines, Electroporation, Gene therapy, In vivo delivery, Inflammation, Animal models