Background: Arterial stiffness is a determinant of cardiovascular disease in end stage renal disease. Hemodialysis patients may develop anti-platelet factor 4/heparin antibody (PF4-H Ab) because of heparin treatment in dialysis. We tested whether PF4-H Ab was associated with progression of arterial stiffness in a 3-year follow-up. Methods: We enrolled 74 hemodialysis patients and studied their clinical, biochemical and arterial stiffness measurement with brachial-ankle pulse wave velocity (baPWV) over 3 years. Baseline and changes in baPWV after 3 years (baPWV) were collected and compared with related clinical and biochemical parameters. PF4-H Ab was evaluated by the enzyme-linked immunosorbent assay and titer  0.4 was defined to have PF4-H Ab. Results:We found a positive PF4-H Ab status in 25 of 74 patients.Mean baPWVwas 16.1  3.8 (m/s) at baseline and 17.6  4.0 (m/s) after 3 years. Mean baPWV was 3.4  2.2 (m/s) in the PF4-H Ab positive group, and 0.6  1.2 (m/s) in the PF4-H Ab negative group. Baseline baPWV was only significantly associated with age ( = 0.49, p < 0.01). baPWV was significantly different between the PF4-H Ab positive and negative groups (p < 0.01). In multivariate regression analysis, only PF4-H Ab was positively associated with baPWV ( = 0.71, p < 0.01). Conclusions: Our study concluded that PF4-H Abwas associated with progression of arterial stiffness in hemodialysis patients.