The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of nTreg, but it cannot be used to isolate cells for functional studies. In this study, we compared four staining profiles of Treg, including CD4⁺CD25^high T cells, CD4⁺CD39⁺ T cells, CD4⁺CD73⁺ T cells, and CD4⁺CD25⁺CD127^low/− T cells. We found that CD4⁺CD25⁺CD127^low/− T cells expressed the highest level of Foxp3 and had the strongest correlation with CD4⁺CD25⁺Foxp3⁺ T cells, the accepted identifying characteristics for “real” nTreg cells. Moreover, functional data showed that CD4⁺CD25⁺CD127^low/− T cells could effectively suppress the proliferation of CD4⁺CD25⁻ T cells, suggesting that compared with the other three populations, CD4⁺CD25⁺CD127^low/− T cells best fit the definition of naturally occurring regulatory T cells in human peripheral blood. Finally, we showed that CD4⁺CD25⁺CD127^low/− can be used to quantitate Treg cells in individuals with systemic lupus erythematosus supporting the use of CD4⁺CD25⁺CD127^low/− to identify human Treg cells.