Differential Proteomic Analysis Of The SPF Chicken Spleen Tissue Infected With The Reticuloendtheliosis Virus

Reticuloendtheliosis is caused by Reticuloendtheliosis virus （REV）, which is a poultry pathologicalsyndrome characterized by dwarf syndrome, acute reticulum cell tumors, lymph and other tissueschronic tumor. By now, Reticuloendtheliosis has been world-wide distribution and the thirdimmunosuppressive disease following the Marek’s Disease, Avian Leukosis. The infection rate of theREV had reached20%-30%in China, and brought about significant economic losses.Immunosuppression plays an important role in Reticuloendtheliosis occurrence and development.Proteins are the direct execution of the activities of life, so the interactions between virus and hostwould be presented by the differences in protein expression profile. Studying and screening the hostdifferentially expressed proteins due to virus infection will help elucidate the molecular mechanisms ofthe immunosuppression. Spleen is the main target organ of the REV and can effectively reflect theinteraction relationships between REV and host.54chickens were randomly divided into2groups and housed in the isolated facilities. One group（27）chickens were challenged with104.6TCID50of REV（HLJ07I strain） at3day of age, the rest（27）were kept as uninfected controls. REV-infected and uninfected control chickens were kept in separateunits with similar environmental conditions. At7,14and21dpi,9chickens that were randomly selectedfrom each group were euthanized using CO₂inhalation. At necropsy, a portion of spleen was collectedfrom each chicken kept at-70℃until further processing（any3spleens were a sample pool in eachgroup）.Through the optimization of the two-dimensional electrophoresis conditions, we established thetwo-dimensional electrophoresis method for the detection of differentially expressed proteins. Here60proteins were successfully identified by MALDI-TOF/TOF MS technology. The differentially expressedproteins involve in cytoskeleton, macromolecular metabolism, stress response, signal transduction, cellproliferation and apoptosis, et al.12differentially expressed proteins: Hsp70, PDIA3, Rho-GDI,PRPF19, RPSA, ANXA6, ACTG5, Hsp70, Ferritin H, Serum precursor, LAMB2and VCP were verifiedby Real-time PCR technology at the transcriptional level, and the results were basically consistent withthe results of2D gel electrophoresis. Protein Rho-GDI was verified by Western Blot successfully.The differentially expressed proteins data of the REV-infected and the uninfected control spleen ofSPF chickens were obtained successfully. We discussed the potential role of these proteins on theinteractions between REV and host. The present findings provide a basis for further studies aimed atelucidation of the role of these proteins in REV interactions with its host.