A. A. Karyadi et al. Int J App Pharm, Vol 13, Special Issue 4, 2021, 106-111 1 st Bandung International Teleconference on Pharmacy (BITP), 2021 | 106 SINGLE-NUCLEOTIDE POLYMORPHISM OF TNFSF4 (RS2205960) OF SYSTEMIC LUPUS ERYTHEMATOSUS PATIENTS IN BANDUNG, INDONESIA Original Article ANGELA APRILIA KARYADI 1 , ERICA WILLY 1 , GABRIEL BAGUS KENNARDI 2 , MUHAMMAD SYAWAL PRATAMA 3 , DESI RESKI FAJAR 1 , LANIYATI HAMIJOYO 2,4 , RIEZKI AMALIA 3,5 , MELISA INTAN BARLIANA 1,5 1 Department Biological Pharmacy, Faculty of Pharmacy, Padjadjaran University, Jl. Raya Bandung-Sumedang KM 21, Jatinangor, 45363, Indonesia, 2 Immunology Study Center, Faculty of Medicine, Padjadjaran University, Jl. Raya Bandung-Sumedang KM 21, Jatinangor, 45363, Indonesia, 3 Department of Pharmacology and Clinical Pharmacy, Padjadjaran University, Jl. Raya Bandung-Sumedang KM 21, Jatinangor, 45363, Indonesia, 4 Department of Internal Medicine Faculty of Medicine, Padjadjaran University, Hasan Sadikin Hospital, Jl. Pasteur 38 Bandung, 40161, Indonesia, 5 Center of Excellence in Higher Education for Pharmaceutical Care Innovation, Padjadjaran University, Jl. Raya Bandung-Sumedang KM 21, Jatinangor, 45363, Indonesia Email: melisa.barliana@unpad.ac.id Received: 08 Aug 2021, Revised and Accepted: 15 Aug 2021 ABSTRACT Objective: This study aimed to determine the genotype distribution of the TNFSF4 (Tumor Necrosis Factor Superfamily 4) gene rs2205960 in Systemic Lupus Erythematosus (SLE) patients in Bandung, West Java, Indonesia. Methods: This was a cross-sectional study; 84 genomic DNA samples were amplified, electrophoresed, then analyzed by DNA sequencing. Results: The genotype distribution of the TNFSF4 gene rs2205960 in SLE patients showed that from 84 DNA samples, 55 patients are GG (65.48%), 25 patients are GT (29.76%), and 4 patients are TT (4.76%). Conclusion: Results indicate that SLE patients in Bandung have a genotype distribution of the TNFSF4 rs2205960 gene that fulfills the Hardy- Weinberg equilibrium. Keywords: Bandung, Single-nucleotide polymorphism, RS2205960, Systemic lupus erythematosus, TNFSF4 © 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2021.v13s4.43827 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Systemic Lupus Erythematosus (SLE) is a chronic inflammatory autoimmune disease with various clinical manifestations affecting multiple organ systems [1]. SLE patients experience a loss of self- tolerance due to an abnormal immunological function, which leads to the formation of immune complexes that can damage tissues and characterized by the production of autoantibodies, activation of the complement system, and the involvement of genetic and environmental components [1, 2]. The cause of SLE is very multifactorial, between various genetic and environmental factors that contribute to disease susceptibility [3]. In recent years, many studies have shown the relationship between polymorphisms of several genes with SLE susceptibility, one of which is the Tumor Necrosis Factor Superfamily 4 (TNFSF4) gene. This gene is located in chromosome 1 at position 25 (1q25). The TNFSF4 gene encodes OX40L, a ligand for the OX40 receptor, which is a member of the tumor necrosis factor (TNF) superfamily that is expressed on antigen-presenting cells, such as B cells, dendritic cells, macrophages, mast cells, endothelial vascular cells, and natural killer cells [4–8]. Single-nucleotide polymorphism (SNP) of the TNFSF4 gene rs2205960 has the most significant association in the TNFSF4 region in SLE patients, according to Genome-Wide Association Studies (GWAS) conducted by Chang et al. (2019). This intronic SNP is located 38.6 kb upstream of TNFSF4 and implicated as a cis-acting factor for transcription [8–10]. SNP rs2205960 (G>T) causes an increase in OX40L expression, which influences the development of SLE disease. Overexpression of OX40L may increase the stimulation of CD4 + T helper effector cells and increase the response of follicular T helper cells whose function is to help B cells proliferate and form antibodies causing inflammation and autoimmunity in specific organs [11–13]. The OX40L signaling suppresses the generation and function of interleukin-10 (IL10)-producing CD4 + type 1 regulatory T cells, which play an important role in maintaining peripheral resistance [14]. According to the meta-analysis conducted by Wang et al. (2019), the TT genotype of the TNFSF4 rs2205960 increases the risk of SLE with a significant degree of association [11]. Another study stated that the T allele of TNFSF4 rs2205960 had a significant association in SLE patients from Asian, Caucasian, Hispanic, and Mexican races [10, 12– 19]. These results showed the influence of race and ethnicity on SNP. However, research on TNFSF4 gene polymorphisms, especially rs2205960 in SLE patients in Indonesia, has not been reported. Based on this background, this study identified the gene polymorphism of TNFSF4 rs2205960 and analyze the allele and genotype frequencies in SLE patients in Bandung using the direct sequencing method. MATERIALS AND METHODS Subjects Blood samples from 84 patients were collected from the Hasan Sadikin Hospital in Bandung, Indonesia. Patients who were included in this study were female patient’s aged 18-65 y diagnosed with SLE. Patients were willing to participate in the research and agreed to sign the informed consent. Patients excluded from this study were SLE patients with incomplete treatment data or medical records that could not be traced and patients who were not willing to participate in the study. This study protocol was approved by the Health Research Ethics Committee Universitas Padjadjaran (reference no. 128/UN6. KEP/EC/2020). DNA isolation DNA isolation was done using Geneaid TM DNA Isolation Kit (Geneaid TM , New Taipei City, Taiwan), which is based on the precipitation method. Extracted DNA samples were then stored at- 20 °C until further use. International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 13, Special Issue 4, 2021