Naïve perforin-deficient BALB/c mice survive while vaccinated PKO mice containing virus-specific memory CD8+ T cells rapidly

succumb to lymphocytic choriomeningitis virus (LCMV) infection. Thus, vaccination converts a nonlethal persistent infection into a fatal disease mediated by virus-specific memory CD8+ T cells. Here, we determine the extent to which vaccination-induced mortality in PKO mice following LCMV challenge is due to differences in vaccine modalities, the quantity or epitope specificity of memory CD8+ T cells. We show that LCMV-induced mortality in immune PKO mice is independent of vaccine modalities and that the starting number of memory CD8+ T cells specific to the immunodominant epitope NP118-126 dictates the magnitude of secondary CD8+ T-cell p38 MAPK activity expansion, the inability to regulate production of CD8+ T-cell-derived IFN-γ,

and mortality in the vaccinated PKO mice. Alisertib Importantly, mortality is determined by the epitope specificity of memory CD8+ T cells and the associated degree of functional exhaustion and cytokine dysregulation but not the absolute magnitude of CD8+ T-cell expansion. These data suggest that deeper understanding of the parameters that influence the outcome of vaccine-induced diseases would aid rational vaccine design to minimize adverse outcomes after infection. Following infection or immunization, Ag-specific CD8+ T cells undergo vigorous expansion in numbers and differentiation into effector cells [[1-6]] that are capable of perforin-dependent cytolysis and production of cytokines such as IFN-γ and TNF [[7]]. Tight Janus kinase (JAK) regulation of cytolysis and cytokine production by effector and memory CD8+ T cells is thought to minimize immunopathology [[8]]. CD8+ T-cell responses to infection can be associated with lethal immunopathology

as evidenced by uniform, perforin-dependent mortality after intracranial injection of mice with lymphocytic choriomeningitis virus (LCMV) [[9, 10]]. In addition to its cytotoxic function in the granule exocytosis effector pathway in CD8+ T cells and NK cells [[11]], perforin has also been shown to regulate other aspects of the Ag-specific CD8+ T-cell response, including the degree of proliferative expansion in a bacterial infection [[12]], exhaustion in chronic viral infection [[13, 14]], and survival of CD8+ T cells in models of graft-versus-host disease [[15]]. However, the precise role of perforin in regulating these aspects of the CD8+ T-cell response is still unclear. In particular, the role of perforin in regulating the secondary CD8+ T-cell response to infection has not been well characterized. Additionally, perforin-deficient (PKO) mice serve as a clinically relevant model for the human disease, familial hemophagocytic lymphohistiocytosis (FHL) [[16-19]].