Overall, these data indicated that combination therapy further enhanced Th1-dominant antitumor immunity induced by cryo-thermal therapy and inhibited the differentiation of Tregs, Th17 and Tfh, as well as CD4+ T exhaustion, which contributed to a durable antitumor immune response, leading to a significant improvement in the survival rate of the 4T1 breast cancer-bearing mice. This evidence concerns the gene CD4 and breast cancer.