Human Embryonic Stem Cell Lines Model Experimental Human Cytomegalovirus Latency

Abstract

ABSTRACT Herpesviruses are highly successful pathogens that persist for the lifetime of their hosts primarily because of their ability to establish and maintain latent infections from which the virus is capable of productively reactivating. Human cytomegalovirus (HCMV), a betaherpesvirus, establishes latency in CD34+ hematopoietic progenitor cells during natural infections in the body. Experimental infection of CD34+ cells ex vivo has demonstrated that expression of the viral gene products that drive productive infection is silenced by an intrinsic immune defense mediated by Daxx and histone deacetylases through heterochromatinization of the viral genome during the establishment of latency. Additional mechanistic details about the establishment, let alone maintenance and reactivation, of HCMV latency remain scarce. This is partly due to the technical challenges of CD34+ cell culture, most notably, the difficulty in preventing spontaneous differentiation that drives reactivation and renders them permissive for productive infection. Here we demonstrate that HCMV can establish, maintain, and reactivate in vitro from experimental latency in cultures of human embryonic stem cells (ESCs), for which spurious differentiation can be prevented or controlled. Furthermore, we show that known molecular aspects of HCMV latency are faithfully recapitulated in these cells. In total, we present ESCs as a novel, tractable model for studies of HCMV latency. IMPORTANCE Human cytomegalovirus (HCMV) is a significant human pathogen that is known for causing birth defects, blindness in AIDS patients, and organ transplant rejection. The ability of HCMV to cause disease is dependent upon its capacity to establish and maintain latent infections. Very few of the molecular mechanisms of latency have been elucidated, due in part to the lack of a tractable cell culture model. Here we present embryonic stem cells (ESCs) as a model for HCMV latency, one in which genome maintenance and reactivation could be closely monitored. HCMV establishes latency in ESCs in the same fashion as it does in CD34+ cells, the currently favored in vitro model. Hence, ESCs represent a novel model with unique properties, such as the ability to be genetically manipulated and cultured indefinitely in an undifferentiated state, that will facilitate the mechanistic examination of certain aspects of HCMV latency that have proven technically challenging in other model systems.