Adjuvant therapy with IFN is associated with a significant improvement in disease-free survival (DFS) and overall survival (OS).43, 44 In our study, without any adjuvant therapy, a high level of IFN-γ was boosted after the cryo-thermal therapy, which provides an ideal microenvironment for optional T-cell function.30 Altogether, our studies demonstrated that long-term survival of a murine B16F10 melanoma induced by the cryo-thermal therapy mainly attributed to the induction of antigen-specific CD4+ T cells maintaining melanoma-specific memory immune response. The gene discussed is CD4; the disease is melanoma.