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REPORTS derived from murine gammaherpesvirus68-infected mice. Journal of Virology. 70(9):6516-8. 10. Webster GA, Perkins ND. 1999. Transcriptional cross talk between NFkappaB and p53. Molecular and Cellular Biology. 19(5):3485-95. 11. Wu TT, Usherwood EJ, Stewart JP, Nash AA, Sun R. 2000. Rta of murine gammaherpesvirus 68 reactivates the complete lytic cycle from latency. Journal of Virology. 74(8):3659-67.

Figure 7: Immunoblot of TPA and/or Bay11-treated HE cell lysates. At 48 hours, there is an increase in lytic antigen with TPA treatment (lane 8 versus lane 3). Combination with Bay11 further increases lytic antigen (lane 8 versus lane 10).

sus uninduced samples, showing a better delineation between the latent and lytic state. Therefore, we have continued using this cell line to evaluate optimal reactivation conditions.

Conclusions In this study, we aimed to optimize cell culture conditions to induce lytic replication in MHV68-infected B cell lines. For the latently infected A20-HE cells, TPA-stimulated reactivation was enhanced with the addition of NF-kB inhibitors, supporting our model whereby NF-kB activation promotes latency by repressing lytic gene expression. The S11 and M12 cell lines can also be induced to reactivate; however, because there is a great deal of spontaneous reactivation, they may not be ideal for studying the role of NF-kB in reactivation. In the future, we plan to explore additional cell lines, further validate our findings and identify the viral genes that are regulated by NF-kB signaling.

References 1. Barton E, Mandal P, Speck SH. 2011. Pathogenesis and host control of gammaherpesviruses: lessons from the mouse. Annual Review of Immunology. 29:35197. 2. Brown HJ, Song MJ, Deng H, Wu TT, Cheng G, Sun R. 2003. NF-kappaB inhibits gammaherpesvirus lytic replica-

tion. Journal of Virology. 77(15):8532-40. 3. Forrest JC, Speck SH. 2008. Establishment of B-cell lines latently infected with reactivation-competent murine gammaherpesvirus 68 provides evidence for viral alteration of a DNA damagesignaling cascade. Journal of Virology. 82(15):7688-99. 4. Gradoville L, Kwa D, El-Guindy A, Miller G. 2002. Protein kinase C-independent activation of the Epstein-Barr virus lytic cycle. Journal of Virology. 76(11):5612-26. 5. Keller SA, Hernandez-Hopkins D, Vider J, Ponomarev V, Hyjek E, Schattner EJ, Cesarman E. 2006. NF-kappaB is essential for the progression of KSHV- and EBV-infected lymphomas in vivo. Blood. 107(8):3295-302. 6. Krug, LT, JM Moser, SM Dickerson, and SH Speck. 2007. Inhibition of NFkB activation in vivo impairs establishment of gammaherpesvirus latency. PLoS Pathogens 3(1): e11 7. Krug, LT, CM Collins, LM Gargano, SH Speck. 2009. NF-kB p50 plays distinct roles in the establishment and control of murine gammaherpesvirus 68 latency. Journal of Virology. 83:4732-4748 8. Krug, LT, E Torres-Gonzรกlez, Q Qin, D Sorescu, M Rojas, A Stecenko, SH Speck, AL Mora. 2010. Inhibition of NF-kappa B signaling reduces virus load and gammaherpesvirus-induced pulmonary fibrosis. American Journal of Pathology. 177: 608-621 9. Usherwood EJ, Stewart JP, Nash AA. 1996. Characterization of tumor cell lines The Stony Brook Young Investigators Review, Fall 2011


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Fall 2011  

Fall 2011