Diabetes mellitus (DM) is a health concern because it leads to complications such as retinopathy. Pakistan has 6.9 million DM affected people that will be doubled by 2025. A study was designed to enumerate CD4(+)CD25(+)Treg cells in Pakistani type 2 diabetes mellitus (T2DM) patients. It was a cross-sectional case-control study that included 212 subjects. The subjects having diabetic retinopathy were labeled as Group-I (30 healthy volunteers without diabetes), Group-II (30 T2DM patients without retinopathy) and Group-III (152 T2DM patients with retinopathy). The percentage of CD4+CD25+ Treg cells was determined by Flowcytometry. Comparison of CD4(+)CD25(+)T cells among different groups was not significant and higher percentage of Treg cells was observed in Group-II (3.07%) compared to Group-III (2.88%). Age, gender and duration of diabetes may contribute while percentage of CD4(+)CD25(+)T cells and Treg cells were not associated with the development of DR in T2DM.