Since PD-1 expression was enhanced on all CD4+T cell subsets on both chronic HCV infected patients and healthy controls, it is necessary and informative to compare the ratio of TEM-Tregs to naive-Tregs and of TEM-Tregs to TCM-Tregs regarding to PD-1 MFI between CHC patients and HCs. Here, CD4 is linked to cryohydrocytosis.