IFN-γ Critically Enables the Intratumoural Infiltration of CXCR3(+) CD8(+) T Cells to Drive Squamous Cell Carcinoma Regression

SIMPLE SUMMARY: Cutaneous squamous cell carcinoma (SCC) is prevalent in aged individuals and individuals with compromised or weakened immune systems, indicating a close association between immune function and SCC control. The aim of our study was to uncover the identity of key immune subsets that mediate SCC control, and to elucidate the mechanistic role of the proinflammatory cytokine Interferon-gamma in this process. We established a SCC regressor model, which we used to determine that: (1) CD8(+) T cells, not CD4(+) T cells or NK cells, are essential for SCC regression; (2) the neutralisation of Interferon-gamma prevents CD8(+) T cell infiltration and SCC regression; (3) CD8(+) T cell migration into SCC critically depends upon Interferon-gamma-induced chemokine expression. Thus, our model can be used to understand the key immune mechanisms involved in SCC regression, which will support targeted investigations into the integrity of these mechanisms in patients with progressive disease. ABSTRACT: Ultraviolet (UV) radiation-induced tumours carry a high mutational load, are highly immunogenic, and often fail to grow when transplanted into normal, syngeneic mice. The aim of this study was to investigate factors critical for the immune-mediated rejection of cutaneous squamous cell carcinoma (SCC). In our rejection model, transplanted SCC establish and grow in mice immunosuppressed with tacrolimus. When tacrolimus is withdrawn, established SCC tumours subsequently undergo immune-mediated tumour rejection. Through the depletion of individual immune subsets at the time of tacrolimus withdrawal, we established a critical role for CD8(+) T cells, but not CD4(+) T cells, γδ T cells, or NK cells, in driving the regression of SCC. Regression was critically dependent on IFN-γ, although IFN-γ was not directly cytotoxic to SCC cells. IFN-γ-neutralisation abrogated SCC regression, significantly reduced CD8(+) T cell-infiltration into SCC, and significantly impaired the secretion of CXCL9, CXCL10 and CCL5 within the tumour microenvironment. A strong positive correlation was revealed between CXCL10 expression and CD8(+) T cell abundance in tumours. Indeed, blockade of the CXCL10 receptor CXCR3 at the time of tacrolimus withdrawal prevented CD8(+) T cell infiltration and the regression of SCC. Chimeric models revealed an important role for immune cells as producers of IFN-γ, but not as recipients of IFN-γ signals via the IFN-γ receptor. Together, these findings suggest a key role for IFN-γ in driving the expression of chemokines within the tumour environment essential for the destruction of established SCC by CD8(+) T cells.