The present invention provides compositions and methods of use in investigations of the formation of mulitprotein assemblies implicated in disease. Also provided are assays for screening candidate compounds of potential utility in preventing and/or treating such diseases by preventing the assembly of or disrupting the function of multiprotein assemblies.

The present invention relates to nucleic acid sequences that encode hepatitis C viruses (HCV) that are useful in the fundamental research of HCV as well as in the search of a vaccine against HCV. In particular the present invention relates to nucleic acid sequences that comprises HCVs which are capable of expressing said virus when transfected into cells and are capable of infectivity in vivo.

Hepatitis C virus replication at extrahepatic sites has been suggested; however, complete viral replication has only been confirmed in hepatocytes. Here we show that human adipogenic DLK-1.sup.+ stem cells (hADSC) freshly isolated from HCV-infected individuals contained viral transcripts, replication intermediates and viral antigens in vivo, and viral transcripts increased in supernatants upon prolonged ex vivo culture. Furthermore, naive hADSC isolated from HCV () individuals support complete replication of clinical isolates in vitro, and the infection is donor-nonspecific for cells and cross-genotypic for viruses. Viral infection/replication is mediated through CD81, LDL-R, SR-B1, EGFR, Apolipoprotein E, occludin, claudin-1, NPC1L1 and diacylglycerol acetyltransferase-1, and can be inhibited by anti-viral drugs. In addition, the physical properties of hADSC-propagated viral particles resemble clinical isolates more than JFH1/HCVcc, and viruses propagated by in vitro infected hADSC are infectious to primary human hepatocytes. Therefore, hADSC are an in vivo HCV reservoir and represent a novel venue of clinical virus-host interaction. hADSC can also be exploited as a physiologically relevant primary cell culture system to propagate clinical isolates.

The invention provides a cell-free system comprising not more than about 5% wheat germ extract for expressing proteins such as viral proteins and proteins required for viral capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids, are provided. Further provided are methods for expressing proteins such as viral proteins, proteins required for capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids using a cell-free system comprising not more than about 5% wheat germ extract. Further provided are methods to assay for compounds that modulate viral protein, viral capsid assembly, and assembly of proteins into multiprotein complexes whose disruption can ameliorate bacterial, parasitic, metabolic, oncologic, immunologic, or CNS disease in a cell-free system comprising not more than about 5% wheat germ extract.

The present disclosure provides compositions, methods and kits for internal controls of microvesicle isolations. The compositions, methods and kits can comprise enveloped viruses, including, but not limited to, inactive mouse hepatitis virus (MHV).

A method for preparing an immunogenic composition, which comprises treating HCV pseudoparticles with 2-3,6,8,9 neuraminidase A to generate the immunogenic composition. In addition, the immunogenic compositions are made in vaccines for eliciting an immune response to HCV in a subject.