Comparative Proteome Study On Target Organ Of Chickens Infected With Virulent Infectious Bursal Disease Virus

Infectious bursal disease virus (IBDV), a member of the genus Avibirnavirus within the family Birnaviridae, is the causative agent of infectious bursal disease (IBD). IBDV causes severe immunodeficiency in young chickens by destroying immature antibody-producing B lymphocytes in the bursa of Fabricius (BF), which is followed by bursal atrophy. This renders the animal increasingly susceptible to disease, and decreases the effectiveness of vaccination against infectious diseases. To characterize the host response to IBDV infection, the differential proteomes of bursa of chicken, with and without IBDV infection, were analyzde at different time with two-dimensional gel electrophoresis (2-DE) followed by MALDI-TOF/TOF identification.Optimized two-dimensional gel electrophoresis protocol for isolation of soluble proteins from poultry immune organs.2-DE is a potent method to study protein expression and function in living organisms. It allows a fast overview of changes in cell processes by analysis of the entire protein extracts. However there is little information about the use of 2-DE in avian organs. In this study,2-DE sample preparation methodologies including removal of interfering compounds, protein extraction methods and extraction buffers, were optimized using bursa of Fabricius (BF) of chickens as a model system, when dealing with poultry immune organs. An extraction protocol involving in the extraction buffer IV containing 7M urea,2M thiourea,2%(w/v) CHAPS,50mM DTT,0.2% Bio-Lyte 3/10, 1mM PMSF,20U/ml DNase I,0.25 mg/ml RNase A and combination of sonication and vortex, gave the best 2-DE quality. Compared to non-frozen IPG strips, after IEF, the frozen IPG strips did not result in significant difference in the 2-DE patterns. In addition, this optimized protocol was also successfully applied in 2-DE analysis of spleen and thymus in chicken. Theses data imply that the optimized protocol is a potential useful tool for comparative proteomics analysis of avian immune tissues.Comparative proteomic analysis of infectious bursal disease virus-infect bursal of chicken. In different viral infection, host cells will make a variety of defense response. To understand the interaction between the host and virus at the molecular level, the comparative proteomes of IBDV-infected and mock-infected bursal of chicken at five different time points were analyzed using two-dimensional gel electrophoresis (2-DE) followed by MALDI-TOF/TOF protein identification. The analyses of multiple 2-DE gels indicated that a total of 65 protein spots (corresponding to 54 altered proteins) differentially expressed during IBDV infection, including 12 up-regulated proteins and 42 down-regulated proteins. Database search and identification of functional protein analysis showed that these altered proteins mainly related to cytoskeleton network, stress response, macromolecular biosynthesis, energy metabolism ubi-quitin-proteosome pathway, signal trasduction. These proteins may be involved, through various pathwaym, in the IBDV infection and play different functions. This work effectively provides a wealth of experimental data and useful clues to further reveal the molecular mechanism of IBDV pathogenicity, and provides an important reference for the discovery and development of potential molecular biomarkers.