Evaluation of Ebv Transformation of Human Memory B-Cells Isolated by Facs and Macs Techniques Publisher Pubmed



Sadreddini S¹ ; Jadidiniaragh F^{1, 2} ; Younesi V³ ; Pourlak T^{1, 4} ; Afkham A¹ ; Shokri F^{2, 3} ; Yousefi M^{1, 4} Show Affiliations
Source: Journal of Immunotoxicology Published:2016

Abstract

Abstract: Several studies have been performed to develop effective neutralizing monoclonal antibodies. The Epstein–Barr virus (EBV) can efficiently immortalize B-cells to establish lymphoblastoid cell lines (LCL) and so it has been used extensively for transformation of B-cells to produce and secrete immunoglobulin. The present study addressed the effect of TLR7/8 agonist (R848), feeder cells layer and fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) cell separation methods on the transformation efficiency of antibody-producing memory B-cells. For these studies, the antigen used for analyses of antibody formation was the tetanus neurotoxin (TeNT) derived from Clostridium tetani. The results here showed that employing an HFFF.PI6 feeder cell layer, R848 agonist and FACS-mediated purification of memory B-cells led to increased transformation efficiency. Altogether, the effects of the R848 and the feeder cells provided an efficient method for EBV transformation of human B-cells. Moreover, there was an advantage in using FACS sorting of B-cells over the MACS method in the context of EBV transformation and immortalization of precursors of antigen-specific B-cells. © 2016 Informa UK Limited trading as Taylor & Francis Group.