Spatial distribution of antigen-specific CD8+ T cells in virally infected skin

Abstract

Abstract CD8+ T cells play a critical role in the elimination of viral infections. After epicutaneous infection with vaccinia virus (VACV), virus replicates in skin keratinocytes and antigen drains to the lymph node, priming antigen-specific CD8+ T cells. Once activated, CD8+ T cell migration into the skin occurs independent of cognate antigen recognition. In laboratory mice, which are immunologically naive prior to viral infection, only antigen-specific CD8+ T cells are activated and can migrate into the skin. However, as humans are exposed to many more pathogens than laboratory mice, activated CD8+ cells present in the circulation can be recruited to the site of infection, regardless of their antigen specificity. To model this phenomenon in mice, we adoptively transferred OT-I CD8+ T cells, infected in the footpad with non-replicating virus to activate these T cells, and then infected epicutaneously with recombinant VACV expressing or lacking SIINFEKL (the cognate antigen for OT-I cells). This system allows the robust recruitment of both antigen-specific and non-specific antiviral CD8+ T cells into the skin. Using a combination of flow cytometry and confocal and intravital microscopy, we are currently investigating the spatial distribution of antigen-specific and non-specific CD8+ T cells in the skin. Additionally, we are quantifying cytokine production by these CD8+ T cells under various conditions, including after infection with viruses expressing distinct antigens, in close proximity. These studies should provide additional clarity on the role of antigen in CD8+ T cell spatial distribution in infected peripheral tissues.