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A New Method to Determine Antigen-Specific CD8+ T Cell Activity in Vivo by Hydrodynamic Injection
HIV and Malaria Vaccine Program, The Aaron Diamond AIDS Research Center, Affiliate of the Rockefeller University, New York, NY 10016, USA
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; in revised form: 30 December 2011 / Accepted: 1 January 2012 / Published: 5 January 2012
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
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MDPI and ACS Style
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.
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(1):23-33.
Rai, Urvashi; Huang, Jing; Mishra, Satish; Li, Xiangming; Shiratsuchi, Takayuki; Tsuji, Moriya. 2012. "A New Method to Determine Antigen-Specific CD8+ T Cell Activity in Vivo by Hydrodynamic Injection." Biomolecules 2, no. 1: 23-33.