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Clinical and Vaccine Immunology, July 2006, p. 733-739, Vol. 13, No. 7
1071-412X/06/$08.00+0     doi:10.1128/CVI.00019-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The 3' CCACCA Sequence of tRNA^Ala(UGC) Is the Motif That Is Important in Inducing Th1-Like Immune Response, and This Motif Can Be Recognized by Toll-Like Receptor 3

Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 200032, Shanghai, People's Republic of China,¹ Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853²

Received 10 January 2006/ Returned for modification 24 February 2006/ Accepted 15 May 2006

In this article, the immunogenicity of tRNA and the recognition of tRNA by Toll-like receptors (TLRs) are analyzed. Analyses of the effects of different tRNA^Ala(UGC) fragments (tRNA^Ala1-76 [corresponding to positions 1 through 76], tRNA^Ala26-76, tRNA^Ala40-76, tRNA^Ala62-76, tRNA^Ala1-70, tRNA^Ala26-70, tRNA^Ala40-70, and tRNA^Ala62-70) on the immune responses of hepatitis B surface antigen (HBsAg) were performed with BALB/c mice. Results show that tRNA^Ala1-76, tRNA^Ala26-76, tRNA^Ala40-76, and tRNA^Ala62-76 adjuvants not only induced stronger T helper (Th) 1 immune responses but also cytotoxic-T-lymphocyte (CTL) responses relative to tRNA^Ala1-70, tRNA^Ala26-70, tRNA^Ala40-70, and tRNA^Ala62-70 adjuvants in HBsAg immunization. A deletion of the D loop (tRNA^Ala26-76), anticodon loop (tRNA^Ala40-76), or TC (tRNA^Ala62-76) loop of tRNA^Ala(UGC) does not significantly decrease the adjuvant characteristic of tRNA^Ala(UGC). However a deletion of the 3'-end CCACCA sequence (tRNA^Ala1-70, tRNA^Ala26-70, tRNA^Ala40-70, and tRNA^Ala62-70) of tRNA^Ala(UGC) significantly decreased the adjuvant characteristic in Th1 and CTL immune responses. Moreover, the recognitions of different tRNA^Ala(UGC) fragments by TLR3, TLR7, TLR8, and TLR9 were analyzed. Results show that a deletion of the 3' CCACCA sequence of tRNA^Ala(UGC) significantly decreased the recognition by TLR3. We concluded that the 3' CCACCA sequence of tRNA^Ala(UGC) is the important motif to induce Th1 and CTL responses and this motif can be effectively recognized by TLR3.

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