 | |  | |  | |  | |  | |  | |  | |  |
| IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711 |
Int J Clin Exp Med 2012;5(1):24-33
Original Article
DEC205-DC targeted DNA vaccines to CX3CR1 and CCL2 are potent and limit
macrophage migration
Jimmy Jianheng Zhou*, Yuan Min Wang*, Vincent W S Lee, Richard K S Phoon, Geoff Yu Zhang, Ya Wang, Thian Kui Tan, Min Hu, Lucy
Dongwei Wang, Mitsuru Saito, Andrew Sawyer, David C H Harris, Stephen I Alexander, Anne M Durkan
Centre for Kidney Research, Children’s Hospital at Westmead. Australia. Centre for Transplantation and Renal Research, University of
Sydney at Westmead Millennium Institute. Westmead, NSW 2145, Australia. Kid’s Research Institute, Children’s Hospital at
Westmead, NSW, Australia. *Equal contributors.
Received October 25, 2011; accepted November 6, 2011; Epub November 9, 2011; Published November 30, 2011
Abstract: Received November 2, 2011; accepted December 3, 2011; Epub January 15, 2011; Published January 31, 2011
Abstract: Monocytes utilise a variety of chemokines to traffic to atherosclerotic plaques. CX3C chemokine ligand 1 (CX3CL1 &
Fractalkine) and its receptor CX3CR1 and monocyte chemoattractant protein 1 (CCL2) have been identified as chemokines/receptors
that have an important role in the migration and recruitment of monocytes during the pathogenesis of several inflammatory diseases
including atherosclerosis. DNA vectors containing single chain variable region fragment (scFv) for DC-targeted receptor DEC205 were
cloned with mouse CX3CR1 and CCL2 genes respectively, and vaccinated into C57/BL6 mice weekly for 3 weeks. Induced anti-
CX3CR1 and anti-CCL2 in vaccinated mice was examined by ELISA and Western Blot analysis, while the cellular response was
examined by ELISPOT. The inhibition of chemotaxis of J774 macrophages to Py-4-1 endothelial cells was examined by in vitro
transwell migration assay using serum collected from vaccinated mice. All vaccinated mice generated anti-CX3CR1 and anti-CCL2 Ab
and cellular response by 8 weeks after DNA vaccination. Macrophage migration towards TNF-α activated endothelial cells was
significantly inhibited by serum containing both anti-CX3CR1 or anti-CCL2 Ab from vaccinated mice. These results demonstrate that
DC-targeting of DNA vaccines to self-antigens generates functional immune responses which can inhibit specific key chemotactic
targets. This suggests a potential therapeutic role for chemokine/receptor DNA vaccination in atherosclerosis, where chemotaxis has a
pivotal role in the inflammatory process. (IJCEM1111001).
Keywords: DNA Vaccination, DEC205, macrophage migration, chemokine, CX3CR1, CCL2
Address all correspondence to:
Dr. Anne M Durkan
Centre for Kidney Research
Children’s Hospital at Westmead
Westmead NSW 2145
Sydney, Australia.
Tel: (61-2) 98453106
E-mail: yuanw@chw.edu.au
Original Article
DEC205-DC targeted DNA vaccines to CX3CR1 and CCL2 are potent and limit
macrophage migration
Jimmy Jianheng Zhou*, Yuan Min Wang*, Vincent W S Lee, Richard K S Phoon, Geoff Yu Zhang, Ya Wang, Thian Kui Tan, Min Hu, Lucy
Dongwei Wang, Mitsuru Saito, Andrew Sawyer, David C H Harris, Stephen I Alexander, Anne M Durkan
Centre for Kidney Research, Children’s Hospital at Westmead. Australia. Centre for Transplantation and Renal Research, University of
Sydney at Westmead Millennium Institute. Westmead, NSW 2145, Australia. Kid’s Research Institute, Children’s Hospital at
Westmead, NSW, Australia. *Equal contributors.
Received October 25, 2011; accepted November 6, 2011; Epub November 9, 2011; Published November 30, 2011
Abstract: Received November 2, 2011; accepted December 3, 2011; Epub January 15, 2011; Published January 31, 2011
Abstract: Monocytes utilise a variety of chemokines to traffic to atherosclerotic plaques. CX3C chemokine ligand 1 (CX3CL1 &
Fractalkine) and its receptor CX3CR1 and monocyte chemoattractant protein 1 (CCL2) have been identified as chemokines/receptors
that have an important role in the migration and recruitment of monocytes during the pathogenesis of several inflammatory diseases
including atherosclerosis. DNA vectors containing single chain variable region fragment (scFv) for DC-targeted receptor DEC205 were
cloned with mouse CX3CR1 and CCL2 genes respectively, and vaccinated into C57/BL6 mice weekly for 3 weeks. Induced anti-
CX3CR1 and anti-CCL2 in vaccinated mice was examined by ELISA and Western Blot analysis, while the cellular response was
examined by ELISPOT. The inhibition of chemotaxis of J774 macrophages to Py-4-1 endothelial cells was examined by in vitro
transwell migration assay using serum collected from vaccinated mice. All vaccinated mice generated anti-CX3CR1 and anti-CCL2 Ab
and cellular response by 8 weeks after DNA vaccination. Macrophage migration towards TNF-α activated endothelial cells was
significantly inhibited by serum containing both anti-CX3CR1 or anti-CCL2 Ab from vaccinated mice. These results demonstrate that
DC-targeting of DNA vaccines to self-antigens generates functional immune responses which can inhibit specific key chemotactic
targets. This suggests a potential therapeutic role for chemokine/receptor DNA vaccination in atherosclerosis, where chemotaxis has a
pivotal role in the inflammatory process. (IJCEM1111001).
Keywords: DNA Vaccination, DEC205, macrophage migration, chemokine, CX3CR1, CCL2
Address all correspondence to:
Dr. Anne M Durkan
Centre for Kidney Research
Children’s Hospital at Westmead
Westmead NSW 2145
Sydney, Australia.
Tel: (61-2) 98453106
E-mail: yuanw@chw.edu.au
