Immunostaining of tumor-infiltrating immune-cell-related markers and cytokines in the tumor fractional radiation model of host PD-L1-deficient mice

In this protocol, we describe how to explore immune function in radiation-induced anti-tumor abscopal effect (ATAE) in PD-L1-deficient (PD-L1(−/−)) mice. We detail steps for primary and abscopal tumor model establishment in mice, tumor fractional irradiation scheme, and isolation of single cells from tumor tissues. Furthermore, we describe the analysis of the proportion of immune cells infiltrated in tumors and the specific staining of cytokines released from immune cells to confirm the ability of CD8+ T cells in killing tumors. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2022).(1)