Research article Influence of CTLA-4/CD28/ICOS gene polymorphisms on the susceptibility to cervical squamous cell carcinoma and stage of differentiation in the Polish population Edyta Pawlak a , Lidia Karabon a , Iwona Wlodarska-Polinska b , Anna Jedynak a , Anna Jonkisz c , Anna Tomkiewicz a , Jan Kornafel b , Marcin Stepien b , Agnieszka Ignatowicz b , Arleta Lebioda c , Tadeusz Dobosz c , Irena Frydecka a,d a Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland b Department of Oncology and Gynecological Oncology Clinic, Medical University, Wroclaw, Poland c Department of Forensic Medicine, Medical University, Wroclaw, Poland d Department of Hematology, Blood Neoplastic Diseases and Bone Marrow Transplantation, Medical University, Wroclaw, Poland ARTICLE INFO Article history: Received 22 April 2009 Accepted 5 November 2009 Available online 12 November 2009 Keywords: CTLA-4 CD28 ICOS Gene polymorphism Cervical squamous cell carcinoma ABSTRACT Abnormal expressions of the costimulatory molecules CD28, “inducible co-stimulator” (ICOS), and inhibitory molecule cytotoxic T-lymphocyte antigen– 4 (CTLA-4) lead to disturbances of immune response and entail an increased risk of cancer. This study was undertaken to evaluate the association between the polymorphisms CTLA-4c.49AG, CTLA-4g.319CT, CTLA-4g.*642AT(8_33), CTLA-4g.*6230GA (CT60), CD28c.17+3TC, and ICOSc.1554+4GT(8_15), and susceptibility to CSCC. The association between CTLA-4g.319CT[T] allele and [TT+CT] genotype and susceptibility to CSCC was observed (OR = 1.99, p = 0.003, and OR = 2.07, p = 0.005, respectively). The CTLA-4g.319CT[T] allele and [TT+CT] genotype increased the risk of well-differentiated CSCC by a factor of 3.84 and 4.44 (p = 0.00001, and p = 0.00001, respectively). In patients with moderately differentiated CSCC, a trend toward increased frequency of CTLA-4g.319CT[T] allele and CTLA-4g.319CT[TT+CT] genotype was noticed (OR = 1.68, p = 0.09, and OR = 1.75, p = 0.09, respectively), whereas in low differentiated CSCC such relation was not observed. Both CTLA-4g.*642AT(8_33) [(AT) 8 ]/[(AT) 8 ] homozygote and [(AT) 8 ] allele were overrepresented in CSCC patients in comparison with healthy women (p = 0.004, OR = 1.93, and p = 0.03, OR = 1.41, respectively) but this polymorphism was not related to histologic grade of tumor. CD28c.17+3TC polymorphism was not associated with susceptibility to CSCC in whole group of patients, but CD28c.17+3TC[C] allele and CD28c.17+3TC[CC+TC] genotype were more frequently ob- served among well differentiated CSCC patients compared with controls (OR = 1.89, p = 0.05, and OR = 2.05, p = 0.05, respectively). Other studied polymorphisms were not associated with susceptibility to CSCC and with histologic grade of CSCC. Our results suggest that the CTLA-4 gene is susceptibility gene to CSCC especially to well-differentiated tumor, while association between CD28 gene polymorphism and disease is restricted only to the well-differentiated CSCC.  2010 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. 1. Introduction Cervical squamous cell carcinoma (CSCC) is the third most com- mon cancer diagnosed in women worldwide, with a disproportion- ate share of the mortality associated with this disease occurring in developing countries [1]. T cells play an important role in antitumor immunity, and mol- ecules that mediate regulation of T-cell activity could influence cancer susceptibility. Lymphocytes activation requires two inde- pendent signals. The first, an antigen-specific signal, is sent via the T-cell receptor (TCR) to T cells. The second signal, termed costimu- lation, is dependent on the antigen receptor and is critical to allow full activation [2]. A reduced level of costimulatory CD28 molecule on peripheral blood T lymphocytes in cervical cancer patients was reported by us and others [3,4]. Another member of the CD28 family, namely, the “inducible co-stimulator” (ICOS), is rapidly upregulated on T lymphocytes upon activation [5]. ICOS costimulation induces T-cell proliferation and the production of interleukin (IL)– 4, IL-5, IL-10, interferon (IFN)–, and IFN- [5], but not IL-2 [6]. The inhibitory molecule CTLA-4 (CD152) is minimally expressed on resting T cells and transiently upregulated after stimulation [7]. CTLA-4 blockade leads to enhancement of the immune response * Corresponding author. E-mail address: epawlak@iitd.pan.wroc.pl (E. Pawlak). E.P., L.K., and I.W.-P. contributed equally to this article. Human Immunology 71 (2010) 195–200 Contents lists available at ScienceDirect 0198-8859/10/$32.00 - see front matter  2010 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. doi:10.1016/j.humimm.2009.11.006