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Biomolecules 2012, 2(1), 23-33; doi:10.3390/biom2010023
Article

A New Method to Determine Antigen-Specific CD8+ T Cell Activity in Vivo by Hydrodynamic Injection

1
,
1
,
2
,
1
,
1,†
 and
1,*
1 HIV and Malaria Vaccine Program, The Aaron Diamond AIDS Research Center, Affiliate of the Rockefeller University, New York, NY 10016, USA 2 Michael Heidelberger Division, Department of Pathology, New York University School of Medicine, New York, NY 10016, USA Present address: Otsuka Maryland Medicinal Laboratories, Inc., 9900 Medical Center Drive, Rockville, MD 20850, USA.
* Author to whom correspondence should be addressed.
Received: 25 November 2011 / Revised: 30 December 2011 / Accepted: 1 January 2012 / Published: 5 January 2012
(This article belongs to the Special Issue Feature Papers)
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Abstract

Hydrodynamic tail vein (HTV) delivery is a simple and rapid tail vein injection method of a high volume of naked plasmid DNA resulting in high levels of foreign gene expression in organs, especially the liver. Compared to other organs, HTV delivery results in more than a 1000-fold higher transgene expression in liver. After being bitten by malaria-infected mosquitoes, malaria parasites transiently infect the host liver and form the liver stages. The liver stages are known to be the key target for CD8+ T cells that mediate protective anti-malaria immunity in an animal model. Therefore, in this study, we utilized the HTV delivery technique as a tool to determine the in vivo cytotoxic effect of malaria antigen-specific CD8+ T cells. Two weeks after mice were immunized with recombinant adenoviruses expressing malarial antigens, the immunized mice as well as naïve mice were challenged by HTV delivery of naked plasmid DNA co-encoding respective antigen together with luciferase using dual promoters. Three days after the HTV challenge, non-invasive whole-body bioluminescent imaging was performed. The images demonstrate in vivo activity of CD8+ T cells against malaria antigen-expressing cells in liver.
Keywords: CD8+ T cells; Hydrodynamic tail vein; liver; malaria; plasmid DNA; luciferase; bioluminescence; recombinant adenovirus CD8+ T cells; Hydrodynamic tail vein; liver; malaria; plasmid DNA; luciferase; bioluminescence; recombinant adenovirus
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Rai, U.; Huang, J.; Mishra, S.; Li, X.; Shiratsuchi, T.; Tsuji, M. A New Method to Determine Antigen-Specific CD8+ T Cell Activity in Vivo by Hydrodynamic Injection. Biomolecules 2012, 2, 23-33.

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