Vol 11, Issue 2, 2018 Online - 2455-3891 Print - 0974-2441 ASSOCIATION OF INTERLEUKIN-4 CYTOKINE AND IL-4R α GENE POLYMORPHISM IN β-LACTAM ALLERGIC PATIENTS MANAL M KHADHIM*, DHUHA A HASSAN Department of Medical Microbiology, College of Medicine, University of Al-Qadisiyah, Iraq. Email: manal.kadhim@qu.edu.iq Received: 06 January 2017, Revised and Accepted: 15 January 2017 ABSTRACT Objective: The present study was carried out to estimate the possible role of Interleukin-4 (IL-4)RαQ576R genes polymorphism in the development of immune reaction against penicillin, as well as to study the effect of IL-4 cytokine in regulating allergic reactions. Materials and Methods: Measurement of serum IL-4 concentration was done using enzyme-linked immunosorbent assay technique; IL-4RαQ576R gene polymorphisms were genotyped using polymerase chain reaction-restriction fragment lengths polymorphisms. Comparisons for statistical significance were performed using Mann–Whitney U-test. Results: Comparing with control subjects, there was a significantly increased level of IL-4 (348.53 pg/ml) in penicillin allergic patients versus (284.72 pg/ml) in sera of control subjects. The IL-4RαQ576R alleles were significantly higher in the penicillin allergic individual compared with apparently healthy control subjects. Conclusions: Data study suggested that IL-4 cytokine have some important roles in penicillin hypersensitivity reaction, additionally the IL- 4RαQ576Rgene polymorphisms might involve in modulating of penicillin hypersensitivity. Keywords: β-lactam, Allergy, Genotype, Interleukin-4. INTRODUCTION Beta-lactams(BL) antibiotics are the most widely used in clinical practice worldwide and constitute the most common inducer of adverse drug reactions (ADRs) with an incidence rate of 0.7–10% of all population [1,2]. Adverse effects of BLs are mainly mediated by an immunological mechanism that is known as hypersensitivity reactions (HRs) [3], that account for 6–10% of all ADRs [4], which considered a problem of great concern for regulatory agencies, healthcare system and industry [5]. Undiagnosed beta-lactam allergy is significant and mounting public health issue due to its limitation in drug selection of alternatives, which can be either expensive or more side effects, so misdiagnosis of BLs allergy will increase the risk of frequent hospitalization, the emergence of antibiotic-resistant infection and increase of medical coast [6]. About 10–20% of total population has been marked as allergic to penicillin [7]. However, it has been reported that over 90% of BLs allergic patients were proven to be able to tolerate penicillin antibiotic when administered [8,9]. Penicillin hypersensitivity is mainly related to specific immunological reactions, that being classified as immediately and noon immediate reaction [10]. The immediate reaction usually occurs during 1 h after drug administration and initiated by certain IgE antibody which responsible for immediate anaphylactic reaction sign and symptoms [11]. Non-immediate reaction usually appears within 24 h after BLs administration [12]. Production of IgE is greatly affected by certain cytokines that released by stimulated T-lymphocyte such as Interleukin-4 (IL-4) and IFN-γ [13]. Therefore, excessive release of IL-4 and interferon-γ is believed to be very important in modulating of immediate HR to penicillin [14]. METHODS Subject The current study was carried out during period from second of January 2017 to the end of June 2017, 50 patients with penicillin allergy (24 males and 26 females), 28 of them are allergic/atopic (have an atopic disorder) and 22 only allergic to penicillin, they were recruited from dermatology Outpatients Department of AL-Diwaniya Teaching Hospital. Informed consent obtained from the patients also an ethical approval obtained. Other 50 apparently healthy subjects (13 males and 37 female) they were included as a control group; blood samples were collected from both groups. Skin testing was done with Benzylpenicillin (PG) using SPT on inner of forearms with a concentration of reagent up to 10000 IU/ml (15, 9, 16, 17). The reaction read after 15–20 min it considered +ve when a wheel diameter was ≥3 mm with surrounding areas of erythema. A blood sample was drawn from patients at the tof +ve skin test. Sera were separated from clotting blood at room temperature for 1 h by centrifugation then stored data −30°C until in vitro tests were performed. Another 3 mL of blood was collected in Ethylenediaminetetraacetic acid tubes then stored at 4°Cnfor DNA extractions for detection of IL-4Rα polymorphisms by polymerase chain reaction-restriction fragment lengths polymorphisms (RFLP- PCR) technique. Immunological study Serum level of IL-4 was detected using enzymes linked-immunosorbent assay (ELISA) which purchased from CALBIOTECH (USA), according to instructions of the company. Genotype study Extraction of genomic DNA from blood samples of patients and control done using Genomic DNA mini kit extraction kit (Frozen Blood) Geneaid (USA) and the procedure was done according to company instructions. The extracted blood genomic DNA was checked by using Nanodrop spectrophotometer (THERMO. USA), which measures the concentration of DNA (ng/µL) and checked the purity of DNA through reading absorbance (260/280 nm). RFLP-PCR technique was performed for genotyping and detection of IL-4Rα (IL-4Rα-Q576R) gene polymorphisms in patients with beta-lactam allergy and in healthy control blood samples. This method was carried out according to a © 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2018.v11i2.24701 Research Article