Studies on Canine Epstein-Barr Virus (EBV)-Related Herpesvirus

Autor: Guo-Hua Wu, 吳國華
Rok vydání: 2007
Druh dokumentu: 學位論文 ; thesis
Popis: 95
Epstein-Barr virus (EBV) is a widespread human gammaherpesvirus (Υ-herpesvirus). EBV infection is associated with several malignancies, e.g. B-lymphocyte derived lymphoma, nasopharyngeal carcinoma (NPC), and oral squamous cell carcinoma (OSCC). Presently, EBV is the only lymphocryptovirus (LCV) known to infect humans. Nevertheless, in our previous study, EBV-encoded nonpolyadenylated small RNAs (EBERs) and DNA sequence of BamHI W fragment were first discovered in canine, indicating the possible presence of EBV in canine. In this study, our specific aim was to establish an EBV-transformed canine cell line. We cultured peripheral blood mononuclear cells (PBMCs) from three dogs, 87h, 96h and A195, which had been known previously as positive for BamHI W sequence. During cultivation, small clumps of cell aggregates were observed and the life span of cells was around 8 weeks to 17 weeks, revealing an unusual long life compared with that of ordinary PBMCs. In addition, sequences of BamHI W fragment and latent membrane protein 1 (LMP1) gene were detected by polymerase chain reaction (PCR) as well. Interestingly, oral tumor was later developed in the velum of dog 87h. Based on the pathological analysis, an early stage of canine OSCC (COSCC) was diagnosed. Furthermore, EBER (EBV-encoded small RNA) signal was detected by in situ hybridization (ISH) in the COSCC sections. Primary culture and subsequent passages of these tumor cells, designated as 87hCOSCC, showed morphological transformation. By PCR, we identified BamHI W sequence of EBV in both episomal form of viral DNA and total DNA extracted from 87hCOSCC, as well as in the supernatant of 87hCOSCC culture. These results suggest the maintenance of viral episome and possible release of EBV-related virus particle in the canine OSCC.
Databáze: Networked Digital Library of Theses & Dissertations
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