gamma-herpesviruses are associated with the development of lymphoproliferative disorders and lymphomas, particularly in immunosuppressed individuals. Importantly, a detailed understanding of how chronic virus infection is established and maintained may identify possible targets for interfering with viral persistence in the host. Unfortunately, the human ?-herpesviruses (EBV and KSHV) have very narrow host tropism, thus limiting detailed studies of host-pathogen interactions. MHV68 infection of mice provides a tractable small animal model system for characterizing the role of specific genes in viral pathogenesis and maintenance of chronic infection. We have previously shown that the MHV68 M2 latency-associated antigen plays a critical role in both the establishment of latency and reactivation of MHV68 from infected B cells. Furthermore, our recent studies have shown that M2 induces an IL-10 dependent proliferation and differentiation of primary murine B cells - reminiscent of EBV-driven differentiation of primary human B cells. M2 regulation of cellular IL-10 expression is particularly intriguing given that EBV encodes an IL-10 homolog whose function during virus infection has remained enigmatic. In addition, M2 can drive terminal differentiation of latently infected B cells to plasma cells, with concomitant reactivation of MHV68. Notably, plasma cell differentiation has also been linked to EBV and KSHV reactivation - indicating that virus regulation of plasma cell differentiation is likely a conserved strategy among gammaherpesviruses. Characterization of MHV68 M2 antigen function is expected to reveal cellular pathways that are manipulated by ?-herpesviruses to facilitate persistence and reactivation from B cells, and may ultimately identify targets for the development of anti-viral drugs. We propose the follow 3 specific aims:
Aim 1. Pathogenesis of M2 mutants in vivo: (1.a) Investigate route- and dose-dependent phenotypes of M2 mutants; (1.b) Analysis of M2 mutant phenotypes in mice lacking an adaptive antibody response;and (1.c) Analysis of B cell lines immortalized with MHV68 harboring mutations in the M2 gene.
Aim 2. Regulation of M2 expression: (2.a) Identification of cell types expressing M2 in vivo; (2.b) Map and characterize cis-elements regulating M2 gene expression during latency.
Aim 3. M2 manipulation of B cell signaling pathways: (3.a) Characterize M2 modulation of B cell signaling pathways and cellular gene expression; (3.b) Identification of M2-interacting proteins; (3.c) Characterize M2 induction of IL-10 expression;and (3.d) Characterize M2-mediated events in plasma cell differentiation.

Public Health Relevance

Murine gammaherpesvirus 68 (MHV68) infection of mice provides a tractable small animal model for characterizing chronic gammaherpesvirus infections - which in some settings (e.g., immunocompromised individuals) are associated with the development of lymphoproliferative disorders and lymphomas, as well as several other cancers. The importance of this model is underscored by the narrow host tropism of the human gammaherpesviruses, Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus, and thus the absence of appropriate animal models for these viruses. During the previous funding period we have discovered that expression of the MHV68 latency-associated M2 antigen in B cells drives cellular proliferation, IL-10 expression, antibody class switch recombination and plasma cell differentiation - the latter being linked to virus reactivation. Although the M2 antige is unique to MHV68, the functions of M2 have close parallels in the human gammaherpesviruses - indicating that further characterization of the M2 antigen will provide important new insights into cellular pathways that are targeted by gammaherpesviruses and may ultimately identify targets for the development of antiviral therapies effective against chroni gammaherpesvirus infections.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
AIDS-associated Opportunistic Infections and Cancer Study Section (AOIC)
Program Officer
Beisel, Christopher E
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Emory University
Schools of Medicine
United States
Zip Code
Wakeman, Brian S; Johnson, L Steven; Paden, Clinton R et al. (2014) Identification of alternative transcripts encoding the essential murine gammaherpesvirus lytic transactivator RTA. J Virol 88:5474-90
Rangaswamy, Udaya S; Speck, Samuel H (2014) Murine gammaherpesvirus M2 protein induction of IRF4 via the NFAT pathway leads to IL-10 expression in B cells. PLoS Pathog 10:e1003858
Rangaswamy, Udaya S; O'Flaherty, Brigid M; Speck, Samuel H (2014) Tyrosine 129 of the murine gammaherpesvirus M2 protein is critical for M2 function in vivo. PLoS One 9:e105197
Liang, Xiaozhen; Paden, Clinton R; Morales, Francine M et al. (2011) Murine gamma-herpesvirus immortalization of fetal liver-derived B cells requires both the viral cyclin D homolog and latency-associated nuclear antigen. PLoS Pathog 7:e1002220
Paden, Clinton R; Forrest, J Craig; Moorman, Nathaniel J et al. (2010) Murine gammaherpesvirus 68 LANA is essential for virus reactivation from splenocytes but not long-term carriage of viral genome. J Virol 84:7214-24
Gray, Kathleen S; Forrest, J Craig; Speck, Samuel H (2010) The de novo methyltransferases DNMT3a and DNMT3b target the murine gammaherpesvirus immediate-early gene 50 promoter during establishment of latency. J Virol 84:4946-59
Siegel, Andrea M; Rangaswamy, Udaya Shankari; Napier, Ruth J et al. (2010) Blimp-1-dependent plasma cell differentiation is required for efficient maintenance of murine gammaherpesvirus latency and antiviral antibody responses. J Virol 84:674-85
Liang, Xiaozhen; Collins, Christopher M; Mendel, Justin B et al. (2009) Gammaherpesvirus-driven plasma cell differentiation regulates virus reactivation from latently infected B lymphocytes. PLoS Pathog 5:e1000677
Krug, Laurie T; Collins, Christopher M; Gargano, Lisa M et al. (2009) NF-kappaB p50 plays distinct roles in the establishment and control of murine gammaherpesvirus 68 latency. J Virol 83:4732-48
Herskowitz, Jeremy H; Siegel, Andrea M; Jacoby, Meagan A et al. (2008) Systematic mutagenesis of the murine gammaherpesvirus 68 M2 protein identifies domains important for chronic infection. J Virol 82:3295-310

Showing the most recent 10 out of 16 publications