International Research Journal of Advanced Engineering and Science ISSN (Online): 2455-9024 140 Anis Sulalah, Maimun Z. Arthamin, Chomsin S. Widodo, and Agus Naba, “Analysis of CD4 cell production using PBMC culture after exposure to radiofrequency electromagnetic waves,” International Research Journal of Advanced Engineering and Science, Volume 4, Issue 1, pp. 140-142, 2019. Analysis of CD4 Cell Production Using PBMC Culture after Exposure to Radiofrequency Electromagnetic Waves Anis Sulalah 1 , Maimun Z. Arthamin 2 , Chomsin S. Widodo 3 , Agus Naba 4 1, 3, 4 Department of Physic, Brawijaya University, Malang, Indonesia-65145 2 Department of Clinical Pathology, Brawijaya University, Malang, Indonesia-65145 Abstract— This research was analyze of CD4 cell production in lymphocytes after exposure radiofrequency electromagnetic waves. The sample use lymphocytes from normal human donor blood that have been cultured pheripheral blood mononuclear cell (PBMC). The exposure process is carried out in a box exposure using VSG25A device. The frequencies used were 900 MHz and 1800 MHz. The measurement variable use variation in distance and time. Calculation of CD4 cell production using flowcitometry. The result show that the longer of exposure time, CD4 cell production was increases. The highest CD4 cell changes occured at distance of 2.5 cm (900 MHz frequency). The highest CD4 cell changes occured at distance of 6.25 cm (1800 MHz frequency). Keywords— Radiofrequency, PBMC, lymphocytes, CD4. I. INTRODUCTION The health effects of exposure radiofrequency electromagnetic waves have been widely tested in the last decade, due to increasing intensity of cellphone use as a communication and information media. The frequency of cellphone that are widely used is the GSM frequency (900 MHz and 1800 MHz) [1]. The effect of exposure radiofrequency electromagnetic waves can interfere the body’s cell balance, especially blood cells. Radiofrequency exposure can affect the number of white blood cell, red blood cell and platelet. The changed number of blood cells that significantly can give an indication damage to the body cells [2]. The exposure of GSM radiofrequency, the frequency of 850 MHz can change the number of white blood cells, blood monocyte cell production and lymphocytes are increases, but different results at platelets that are decrease [3]. Radiation exposure of GSM 900 MHz for 2 hours duration caused the production of ROS (Reactive Oxygen Species) in monocytes and lymphocytes are increase significantly. The number of ROS monocytes and lymphocytes that are exposed RF radiation is very different compared to those that are not exposed [4]. Lymphocytes are a large percentage of white blood cell components. Lymphocytes have an important role to provide information about the condition of cells that are still healthy or that have been damaged or dead. Components of lymphocytes consist of T cells, B cells and NK (Natural Killer). Lymphocyte cells provide information about dysregulation of the immune system in the body. T cells can produce cytokines. If the production of cytokines decreases, will affect at activity of cell production in the body [5]. The lymphocyte component in T cells is CD4. CD4 cell counts can be an indication of the stability immune response in the body. Amino acids are CD4 components obtained from the body's mechanism. CD4 cells function as senders of signals to antibodies when there are foreign objects that enter the body [6]. Peripheral Blood Mononuclear Cell (PBMC) is white blood cell that has a rounded single core, found in lymphocyte cells and monocyte cells. These cells are important components of the immune system that are involved in humoral and cellular immunity. Mononuclear cells are widely used in clinical research and applications such as in the fields of microbiology, virology, oncology, vaccine development, transplantation biology, regenerative biology, and toxicology [7]. CD4 is a part of lymphocytes which indicates the stability of cell conditions, so it must be further investigated how RF radiation affects at CD4 cell production in lymphocytes. II. RESEARCH METHOD This research was used lymphocytes from adult human who does not have abnormalities in blood cells as the sample. Donor blood obtained from ± 24 people with male and female gender. The frequencies used were 900 MHz and 1800 MHz. The time variations was used 15 minutes, 30 minutes, 45 minutes and 60 minutes. The distance variation that used at 900 MHz was 0 cm, 2.5 cm, and 5 cm. The distance variation that used at 1800 MHz was 5 cm, 6.25 cm and 25 cm. The exposure box was used during the measurement process. The exposure process is carried out in exposure box using VSG25A device. Before the isolation process, blood cell was added by EDTA (Ethylenediaminetetraacetic acid) nutrition. Nutrition EDTA to maintain stability in the blood. Blood cells that was given EDTA nutrition was not easily damaged [8]. The mixture of blood and EDTA will be placed in a centrifuge tube that has been filled with Ficoll-Hypaque liquid (Fig. 1). Parts of the blood that have a higher density than the Ficoll-Hipaque will pass through the Ficoll layer and parts of the blood (lymphocytes and monocytes). Parts of blood that have a lower density than Ficoll-Hypaque will accumulate in the plasma. The sample in Fig. 1 was centrifuged at room temparature which had a speed 1000 rpm for 30 minutes that resulting 4 layers (Fig. 2).