At the same time, the expression of PD-1, Lag-3, CTLA-4, Tim-3, and Granzyme B on Tregs from cryo-thermal treated mice was decreased, and the expression of perforin, T-bet and IFN-γ in Tregs was not different from that in Tregs from tumor-bearing control mice (Figure 6C–E). This evidence concerns the gene PRF1 and neoplasm.