Biphasic and cardiomyocyte-specific IFIT activity protects cardiomyocytes from enteroviral infection

Viral myocarditis is a serious disease, commonly caused by type B coxsackieviruses (CVB). Here we show that innate immune protection against CVB3 myocarditis requires the IFIT (IFN-induced with tetratricopeptide) locus, which acts in a biphasic manner. Using IFIT locus knockout (IFITKO) cardiomyocytes we show that, in the absence of the IFIT locus, viral replication is dramatically increased, indicating that constitutive IFIT expression suppresses CVB replication in this cell type. IFN pre-treatment strongly suppresses CVB3 replication in wild type (wt) cardiomyocytes, but not in IFITKO cardiomyocytes, indicating that other interferon-stimulated genes (ISGs) cannot compensate for the loss of IFITs in this cell type. Thus, in isolated wt cardiomyocytes, the anti-CVB3 activity of IFITs is biphasic, being required for protection both before and after T1IFN signaling. These in vitro findings are replicated in vivo. Using novel IFITKO mice we demonstrate accelerated CVB3 replication in pancreas, liver and heart in the hours following infection. This early increase in virus load in IFITKO animals accelerates the induction of other ISGs in several tissues, enhancing virus clearance from some tissues, indicating that-in contrast to cardiomyocytes-other ISGs can offset the loss of IFITs from those cell types. In contrast, CVB3 persists in IFITKO hearts, and myocarditis occurs. Thus, cardiomyocytes have a specific, biphasic, and near-absolute requirement for IFITs to control CVB infection. Author summary Viruses can infect the heart, causing inflammation-termed myocarditis-which is a serious, and sometimes fatal, disease. One way to combat the infection is by stimulating our immune system, encouraging it to fight the virus. However, the treatment that is currently used revs up many different parts of our immune system, including some that play little or no role in clearing the virus, and this wide-ranging activation increases the risk of potentially-harmful side effects. We want to identify the parts of the immune system that fight virus infections of the heart, so that we can improve the treatment of viral myocarditis by selectively stimulating only those immune responses, thereby retaining the benefit of treatment (i.e., clearing the virus) while reducing its cost (i.e. lowering the risk of harmful side effects). In this paper, we demonstrate that a family of proteins called IFITs play a role in protecting many tissues against these infections, but are particularly important in heart muscle cells, in which they are indispensable. Thus, IFITs represent a possible target for the treatment of viral myocarditis.