Sustained CD8+ T Cell Memory Inflation after Infection with a Single-Cycle Cytomegalovirus

Abstract

Cytomegalovirus (CMV) is a β-herpesvirus that establishes a lifelong latent or persistent infection. A hallmark of chronic CMV infection is the lifelong persistence of large numbers of virus-specific CD8+ effector/effector memory T cells, a phenomenon called “memory inflation”. How the virus continuously stimulates these T cells without being eradicated remains an enigma. The prevailing view is that CMV establishes a low grade “smoldering” infection characterized by tiny bursts of productive infection which are rapidly extinguished, leaving no detectable virus but replenishing the latent pool and leaving the immune system in a highly charged state. However, since abortive reactivation with limited viral gene expression is known to occur commonly, we investigated the necessity for virus reproduction in maintaining the inflationary T cell pool. We inhibited viral replication or spread in vivo using two different mutants of murine CMV (MCMV). First, famcyclovir blocked the replication of MCMV encoding the HSV Thymidine Kinase gene, but had no impact on the CD8+ T cell memory inflation once the infection was established. Second, MCMV that lacks the essential glycoprotein L, and thus is completely unable to spread from cell to cell, also drove memory inflation if the virus was administered systemically. Our data suggest that CMV which cannot spread from the cells it initially infects can repeatedly generate viral antigens to drive memory inflation without suffering eradication of the latent genome pool. Cytomegalovirus (CMV) establishes life-long, asymptomatic infections in healthy people. Ongoing immune surveillance prevents viral disease but also results in the accumulation of large numbers of virus-specific T cells. The mechanisms by which the virus persists while stimulating such strong immune responses are unknown. We and others had hypothesized that periodic viral replication and spread to neighboring cells allowed CMV to replenish the pool of infected cells while stimulating virus-specific T cells to accumulate. In this manuscript, we have tested this model by blocking the replication or spread of murine cytomegalovirus (MCMV) and found, surprisingly, that accumulation of virus-specific T cells occurs independently of viral replication. Moreover, these T cells developed the terminal differentiated phenotype that is indicative of repeated antigenic stimulation. Thus, these data suggest that CMV can remain active and continuously stimulate the immune system, while avoiding immune-mediated clearance, without the capacity to spread from cell to cell.