ABSTRACT
Hepatitis C virus (HCV) represents a major health burden with more than 170 million individuals currently infected worldwide, equaling roughly 3% of the world's population. HCV preferentially infects hepatocytes and is able to persist in up to 70% of infected individuals. It is estimated that up to 30% of chronically infected individuals will go on to develop progressive liver disease as a result of HCV infection, making the virus the leading cause of liver transplantation in the world. Currently there is no vaccine for HCV. In this study, we have taken a multi-step approach to develop a novel genotype 1a/1b consensus HCV NS3/NS4A DNA vaccine able to induce strong cellular immunity. We show that this construct is able to induce strong anti-NS3/NS4A T cell responses in C57BL/6 mice, as well as, in Rhesus macaques. Our data suggest that DNA vaccines encoding HCV proteins NS3/NS4A merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.
Subject(s)
Carrier Proteins/immunology , Hepacivirus/genetics , Hepacivirus/immunology , Immunity, Cellular/immunology , Viral Nonstructural Proteins/immunology , Viral Proteins/immunology , Amino Acid Sequence , Animals , Cell Separation , Electroporation , Enzyme-Linked Immunosorbent Assay , Epitope Mapping , Fluorescent Antibody Technique , Genotype , Immunization , Interferon-gamma/biosynthesis , Interferon-gamma/genetics , Intracellular Signaling Peptides and Proteins , Macaca mulatta , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Spleen/cytology , Vaccines, DNA/immunologyABSTRACT
Selected potent TRPV1 agonists (1-6) have been modified by 5- or 6-halogenation on the aromatic A-region to analyze their effects on potency and efficacy (agonism versus antagonism). The halogenation caused enhanced functional antagonism at TRPV1 compared to the corresponding prototype agonists. The analysis of SAR indicated that the antagonism was enhanced as the size of the halogen increased (I>Br>Cl) and when the 6-position was halogenated. Compounds 23c and 31b were found to be potent full antagonists with K(i) (as functional antagonist)=23.1 and 30.3 nM in rTRPV1/CHO system, respectively.