Human cytomegalovirus-triggered necroptosis is suppressed by sequestration of MLKL in the nucleus of infected monocytes

The systemic spread of human cytomegalovirus (HCMV) is associated with severe morbidity and mortality in immunocompromised and immunonaïve patients. Hematogenous dissemination of HCMV to different organ sites is facilitated by peripheral blood monocytes. Circulating monocytes have a short lifespan due, in part, to their intrinsic biological programming to initiate caspase-8-mediated apoptosis upon entry into the circulation from the bone marrow. We previously reported that HCMV extends the lifespan of infected monocytes by blocking procaspase-8 cleavage, yet the precise viral mechanism responsible for suppressing caspase-8 activity remains unknown. Here, we demonstrate that HCMV entry into monocytes rapidly increases the abundance of the anti-apoptotic cellular FLICE-like inhibitory protein long (cFLIP_L), which prevents procaspase-8 cleavage into active caspase-8. However, others have demonstrated that inhibition of caspase-8 opens a “trapdoor” cell death response termed necroptosis. Accordingly, we found that increased levels of cFLIP_L, along with a co-stimulatory signal from Toll-like receptor 3, activate the receptor-interacting protein kinase 3 responsible for initiating necroptosis. Despite the triggering of the necroptotic cascade within infected monocytes, the final execution of this death pathway is thwarted by nuclear sequestering of mixed lineage kinase domain-like pseudokinase, the executioner of necroptosis. Together, our data reveal a multitude of countermeasures employed by HCMV to obstruct cellular antiviral death responses within infected monocytes.