This is a competing renewal of an ROl grant with the long term goal of understanding how Epstein-Barr virus (EBV) gene expression is regulated during immortalizing latency. EBV establishes a life-long infection within the infected host, and is closely associated with the development of endemic Burkitt's lymphoma, nasopharyngeal carcinoma, 30-50 percent of Hodgkin's disease, and nearly half of the lymphomas that arise in immunosuppressed patients. Notably, EBV infection of peripheral resting B cells results in growth transformation resulting, ex vivo, in the generation of immortalized lymphoblastoid cell lines. Based on recent analyses of EBV infection in seropositive individuals, it seems likely that the ability of EBV to drive B cell proliferation, and subsequent differentiation, plays an important role in the dissemination of virus infected B cells and the establishment of a long-lived latency reservoir in memory B cells (in which there is very limited viral gene expression). Understanding how EBV regulates viral gene expression during immortalizing latency may ultimate provide strategies for interfering with this phase of the virus life cycle, which could interfere with the establishment of latency in the memory B cell compartment. Within the scope of this proposal, we will continue to focus our analyses on identifying and characterizing cis-elements involved in regulated EBNA gene expression during the immortalizing latency program of EBV.
Aim 1. Generation of a cottontop marmoset LCL immortalized with a packaging defective and replication null EBV to serve as an inducible reservoir for non-immortalizing EBV mutants.
Aim 2. Generation and characterization of EBV harboring mutations in cis-elements thought to be involved in regulating EBNA gene transcription in EBV immortalized lymphoblastoid cell lines.
Aim 3. Analysis of the requirements for Wp activity during initial stages of infection of primary B cells.
Aim 4. Analysis of Wp methylation during the establishment and maintenance of immortalizing latency in B cells - role of methylation in regulating Wp activity in lymphoblastoid cell lines.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA043143-20
Application #
7046107
Study Section
Virology Study Section (VR)
Program Officer
Wong, May
Project Start
1986-07-01
Project End
2007-03-31
Budget Start
2006-04-01
Budget End
2007-03-31
Support Year
20
Fiscal Year
2006
Total Cost
$278,107
Indirect Cost
Name
Emory University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
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Evans, Andrew G; Moorman, Nathaniel J; Willer, David O et al. (2006) The M4 gene of gammaHV68 encodes a secreted glycoprotein and is required for the efficient establishment of splenic latency. Virology 344:520-31
Allen 3rd, Robert D; Dickerson, Shelley; Speck, Samuel H (2006) Identification of spliced gammaherpesvirus 68 LANA and v-cyclin transcripts and analysis of their expression in vivo during latent infection. J Virol 80:2055-62
Herskowitz, Jeremy; Jacoby, Meagan A; Speck, Samuel H (2005) The murine gammaherpesvirus 68 M2 gene is required for efficient reactivation from latently infected B cells. J Virol 79:2261-73
Willer, David O; Speck, Samuel H (2005) Establishment and maintenance of long-term murine gammaherpesvirus 68 latency in B cells in the absence of CD40. J Virol 79:2891-9
Moser, Janice M; Upton, Jason W; Allen 3rd, Robert D et al. (2005) Role of B-cell proliferation in the establishment of gammaherpesvirus latency. J Virol 79:9480-91

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