[show abstract][hide abstract] ABSTRACT: The development of small molecule inhibitors of hepatitis C virus (HCV) core protein as antiviral agents has been intensively pursued as a viable strategy to eradicate HCV infection. However, lack of a robust and convenient small animal model has hampered the assessment of in vivo efficacy of any antiviral compound.
The objective of this work was to develop a novel method to screen anti-core protein siRNA in the mouse liver by bioluminescence imaging. The inhibitory effect of two shRNAs targeting the highly conserved core region of the HCV genome, shRNA452 and shRNA523, was examined using this method. In the transient mouse model, the effect of shRNA-523 was detectable at as early as 24 h and became even more pronounced at later time points. The effect of shRNA-452 was not detectable until 48 h post-transduction. In a stable mouse model, shRNA523 reduced luciferase levels by up to 76.4±26.0% and 91.8±8.0% at 6 h and 12 h after injection respectively, and the inhibitory effect persisted for 1 day after a single injection while shRNA-Scramble did not seem to have an effect on the luciferase activity in vivo.
Thus, we developed a simple and quantitative assay for real-time monitoring of HCV core protein inhibitors in mice.
PLoS ONE 01/2010; 5(11):e14043. · 3.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: The recognition of human leukocyte antigen (HLA) molecules by specific receptors is a crucial step in the regulation of natural killer (NK) cell function. Killer cell immunoglobulin-like receptor (KIR) 3DS1 is one of the activating receptors of NK cell and is implicated in slowing disease progression in HIV infection. KIR3DS1 play an important role in the outcome of multiple diseases associated with viral infections. In contrast to the inhibitory receptor, much less is known about the ligands of KIR3DS1. In order to achieve a better understanding of the biology of KIR3DS1 and its ligand systems, it is necessary to identify the ligands of KIR3DS1. In this work, we utilized recombinant HLA-B2705 molecules and DsbA-KIR3DS1 fusion protein to monitor the interaction between HLA-B2705 complexes and DsbA-KIR3DS1 using BIAcore 3000 SPR sensor and found that the specific binding between KIR3DS1 and HLA-B2705 existed and the affinity was 6.95x10(-6) mol//L. So we concluded that HLA-B2705 is a possible ligand of KIR3DS1.
Protein and Peptide Letters 12/2009; 17(5):547-54. · 1.99 Impact Factor
[show abstract][hide abstract] ABSTRACT: By bioluminescence imaging and hydrodynamic gene transfer technology, the activities of hepatitis B virus (HBV) promoters and the effects of HBV enhancers on these promoters in mice under true physiological conditions have been assessed. Our studies reveal that either of the two HBV enhancers can stimulate HBV major promoter activity in hepa 1-6 cells (in vitro) and in mouse liver (in vivo), and the enhancer effects on the three promoters (S1, S2 and X promoter) are markedly greater in vivo than in vitro. The two HBV enhancers have no cooperative action on HBV promoters in vitro or in vivo.