High-dimensional characterization of IL-10 production and IL-10 dependent regulation during primary gammaherpesvirus infection

Interleukin (IL)-10 is a potent immunomodulatory cytokine produced by multiple cell types to restrain immune activation. Many herpesviruses use the IL-10 pathway to facilitate infection, but how endogenous IL-10 is regulated during primary infection in vivo remains poorly characterized. Here, we infected mice with murine gammaherpesvirus 68 ({gamma}HV68) and analyzed the production, and genetic contribution, of IL-10 by mass cytometry (cytometry by time-of-flight, CyTOF) analysis.{gamma} HV68 infection elicited a breadth of effector CD4 T cells in the lungs of acutely infected mice, including a highly activated effector subset that co-expressed IFN{gamma}, TNF, and IL-10. By using IL-10 green fluorescent protein (gfp) transcriptional reporter mice, we identified that IL-10 was primarily expressed within CD4 T cells during acute infection in the lungs. IL10gfp expressing CD4 T cells were highly proliferative and characterized by the expression of multiple co-inhibitory receptors including PD-1 and LAG-3. When we analyzed acute{gamma} HV68 infection of IL-10 deficient mice, we found that IL-10 limits the frequency of both myeloid and effector CD4 T cell subsets in the infected lung, with minimal changes at a distant mucosal site. These data emphasize the unique insights that high-dimensional analysis can afford in investigating antiviral immunity, and provide new insights into the breadth, phenotype and function of IL-10 expressing effector CD4 T cells during acute virus infection.\n\nThis work was funded by National Institutes of Health Grants R01 AI121300 and R01 CA168558 (to L.F.v.D.), an American Heart Association National Scientist Development grant (#13SDG14510023), the Crohns and Colitis Foundation of America (#311295), a pilot grant from the Lung, Head and Neck Cancer program within the University of Colorado Cancer Center, and a Career Enhancement Award from the University of Colorado Lung Cancer Specialized Program of Research Excellence (P50CA58187) (all to E.T.C.).\n\nThe Lung, Head and Neck program within the University of Colorado Cancer Center, and the Flow Cytometry Shared Resource, are directly funded through support from the National Cancer Institute Cancer Center Support Grant P30CA046934.\n\nAbbreviations used in this article