Proteomics Of N2a Cells Infected With Rabies Virus

The rabies virus is classified in the Lyssavirus genus of family Rhabdoviridae, one of the twenty virus families. The virion or virus particle of rabies involves a bullet-shaped structure, with a cross-sectional diameter of about 75nm, and a length from 100nm to 300nm, which depends on the strain. A typical rabies virion is composed of a genomic RNA molecule and five distinct structural proteins designated as L, G, N, P (or M1), and M (or M2).Rabies is a viral neuroinvasive disease that causes acute encephalitis (inflammation of the brain) in warm-blooded animals. After the symptoms of rabies appear, the disease is nearly always fatal. Late symptoms are very distinctive, and include anxiety, agitation, delirium, paralysis and wild aggressiveness. The rabies virus survives in widespread, varied, rural fauna reservoirs, with dogs as the principal host. There are an estimated 55,000 human deaths annually from rabies worldwide, with about 35,000 in Asia, around 94% of which transmitted by canines. With the advancements of modern biotechnology, much is known about this disease. But many mechanisms still remain to be unraveled. Many viruses have developed a way during evolution to counteract cells'defense systems, including secretion of inflammatory factors and antivirus cytokines that induce cell programmed death. These interactions between virus and host cells in most cases lead to chronic infections, which involve alteration of host cell physiology and proteins expression level.The past few years have witnessed an unprecedented development of proteomic techniques that enable explorations into early markers and pathogenic mechanisms of viral infections. Much can be revealed by comparing proteomic profile differences on 2D gels (two dimensional electrophoresis gels) between infected and controlled cells. That is why this thesis is dedicated to questing for protein expression regulation after cells are infected with rabies virus vaccine low virulent strain(SRV-9), typical aggressive virulent strain(CVS-11),and street virus virulent strain(BD06), using 2D and proteomic techniques. 41 proteins are found to have at least 2 fold change of expression levels, 29 of which are identified by MALDI-TOF-MS/MS (Matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry), including enzymes, cytoskeleton components signal transduction proteins, energy metabolism proteins, antioxidative stress proteins, apoptosis proteins, nucleic acid metabolite proteins and modifying proteins.The comprehensive profile generated from this study provides a platform for continuing research on the molecular pathogenic mechanisms of the rabies virus infection. These differentially expressed proteins represent core molecules for infections of different strains, which pave way for the unearth of rabies pathogenesis mechanisms in the near future.