TheROP16III-dependent early immune response determines the sub-acute CNS immune response and type IIIToxoplasma gondiisurvival

Abstract

AbstractToxoplasma gondiiis an intracellular parasite that persistently infects the CNS and that has genetically distinct strains which provoke different acute immune responses. How differences in the acute immune response affect the CNS immune response is unknown. To address this question, we used two persistentToxoplasmastrains (type II and type III) and examined the CNS immune response at 21 days post infection (dpi). Contrary to acute infection studies, type III-infected mice had higher numbers of total CNS T cells and macrophages/microglia but fewer alternatively activated macrophages (M2s) and regulatory T cells (Tregs) than type II-infected mice. By profiling splenocytes at 5, 10 and 21 dpi, we determined that at 5 dpi type III-infected mice had more M2s while type II-infected mice had more classically activated macrophages (M1s) and these responses flipped over time. To test how these early differences influence the CNS immune response, we engineered the type III strain to lack ROP16 (IIIΔrop16), the polymorphic effector protein that drives the type III-associated M2 response. IIIΔrop16-infected mice showed a type II-like neuroinflammatory response with fewer infiltrating T cells and macrophages/microglia and more M2s and an unexpectedly low CNS parasite burden. At 5 dpi, IIIΔrop16-infected mice showed a mixed inflammatory response with more M1s, M2s, T effector cells, and Tregs, and decreased rates of infection of peritoneal exudative cells (PECs). These data suggested that type III parasites need the early ROP16-associated M2 response to avoid clearance, possibly by the Immunity-Related GTPases (IRGs), IFN-γ dependent proteins essential for murine defenses againstToxoplasma. To test this possibility, we infected IRG-deficient mice and found that IIIΔrop16parasites now maintained parental levels of PECs infection. Collectively, these studies suggest that, for the type III strain,rop16IIIplays a key role in parasite persistence and influences the sub-acute CNS immune response.Author SummaryToxoplasmais a ubiquitous intracellular parasite that establishes an asymptomatic brain infection in immunocompetent individuals. However, in the immunocompromised and the developing fetus,Toxoplasmacan cause problems ranging from fever to chorioretinitis to severe toxoplasmic encephalitis. Emerging evidence suggests that the genotype of the infectingToxoplasmastrain may influence these outcomes, possibly through the secretion ofToxoplasmastrain-specific polymorphic effector proteins that trigger different host cell signaling pathways. While such strain-specific modulation of host cell signaling has been shown to affect acute immune responses, it is unclear how these differences influence the sub-acute or chronic responses in the CNS, the major organ affected in symptomatic disease. This study shows that genetically distinct strains ofToxoplasmaprovoke strain-specific CNS immune responses and that, for one strain (type III), the acute and sub-acute immune responses and parasite survival are heavily influenced by a polymorphic parasite gene (rop16III).