Cytotoxic T-lymphocyte antigen-4 (CTLA-4) is a cell surface protein, which down-regulates the immune response at CTLA-4/CD28/B7 pathway. We aimed to investigate the influence of the -318C/T, +49A/G, -1661A/G and CT60A/G, and CTLA-4 gene polymorphisms on acute rejection of kidney allograft in Turkish patients. The study design was a case-control study that consists of three groups: Group 1 (n=34) represented the kidney transplant (Ktx) recipients who experienced acute rejection, Group 2 (n=47) was randomly assigned Ktx recipients without acute rejection, and Group 3 (n=50) consisting of healthy volunteers to evaluate the normal genomic distribution. The polymerase chain reaction-restriction fragment length polymorphism technique was used to determine the polymorphisms. Genotype and allele frequencies among three groups denoted similar distributions for +49A/G, -1661A/G, and CT60A/G. Conversely, -318C/T genotype was three times more frequent in the acute rejection group than in the non-rejection group (OR=3.45; 95%CI=1.18-10.1, p=0.015) and two times more frequent than the healthy control group (OR=2.45; 95% CI=0.98 - 6.11, p=0.047). Additionally, having a T allele at -318 position was significantly associated with acute rejection (0.147 vs. 0.043, OR=3.45; 95% CI=1.13-10.56, p=0.02). 318C/T gene polymorphism and T allelic variant were found to be associated with increased acute rejection risk in Turkish kidney allograft recipients.