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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

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Received: 25 November 2011; in revised form: 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 which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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.

AMA 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(1):23-33.

Chicago/Turabian Style

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.



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