en Medical Biotechnology Medical Biotechnology مقاله کامل Full Length/Original Article <span style="font-family:Times New Roman;"><span style="line-height:normal"><b><span style="font-size:10.5pt"><span style="color:#1c1e29">Background:</span></span></b><span style="font-size:10.5pt"><span style="color:#1c1e29"> CD20 is a differentiation-related antigen exclusively expressed on the membrane of B lymphocytes. CD20 amplification is observed in numerous immune-related disorders, making it an ideal target for immunotherapy of hematological malignancies and autoimmune diseases. MAb-based therapies targeting CD20 have a principal role in the treatment of several immune-related disordes and cancers, including CLL. Fc gamma receptors mediate CD20 internalization in hematopoietic cells; therefore, this study aimed to establish non-hematopoietic stable cell lines overexpressing full-length human CD20 antigen as an in vitro model for CD20-related studies. </span></span></span><br> <span style="line-height:normal"><b><span style="font-size:10.5pt"><span style="color:#1c1e29">Methods:</span></span></b><span style="font-size:10.5pt"><span style="color:#1c1e29"> <i>CD20</i> gene was cloned into the transfer vector. The lentivirus system was transfected to packaging HEK 293T cells, and the supernatants were harvested. CHO-K1 cells were transduced using recombinant viruses, and a stable cell pool was developed by the antibiotic selection. <i>CD20</i> expression was confirmed at the mRNA and protein levels. </span></span></span><br> <span style="line-height:normal"><b><span style="font-size:10.5pt"><span style="color:#1c1e29">Results</span></span></b><span style="font-size:10.5pt"><span style="color:#1c1e29">: Simultaneous expression of GFP protein facilitated the detection of CD20-expressing cells. Immunophenotyping analysis of stable clones demonstrated expression of CD20 antigen. In addition, the mean fluorescence intensity was significantly higher in the CD20-CHO-K1 clones than the wild-type CHO-K1 cells. </span></span></span><br> <b><span style="font-size:10.5pt"><span style="line-height:115%"><span style="color:#1c1e29">Conclusion:</span></span></span></b><span style="font-size:10.5pt"><span style="line-height:115%"><span style="color:#1c1e29"> This study is the first report on using second-generation lentiviral vectors for the establishment of a non-hematopoietic cell-based system, which stably expresses full-length human CD20 antigen. Results of stable CHO cell lines with different levels of CD20 antigen are well suited to be<br> used for CD20-based investigations, including binding and functional assays.</span></span></span></span> CD20, immunotherapy, Chinese Hamster Ovary cells, lentivirus 269 279 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-814-3&slc_lang=en&sid=1 Niloufar Mohammadkhani nsr.sbmu@gmail.com 0000-0002-9504-123X No Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Azam Rahimpour rahimpour@sbmu.ac.ir 0000-0002-3296-5881 No Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran Reyhaneh Hoseinpoor reyhanehoseinpour@gmail.com 0000-0002-6173-0459 No Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Masoumeh Rajabibazl rajabi_m@sbmu.ac.ir 0000-0002-9720-5904 Yes Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran