Manutsanun SumonwiriyaNavin HorhthongkhamKovit PattanapanyasatSilawun AmpolReungpung SutthentWannee KantakamalakulMahidol UniversityFaculty of Medicine, Siriraj Hospital, Mahidol University2018-09-132018-09-132009-10-01Journal of Virological Methods. Vol.161, No.1 (2009), 154-160016609342-s2.0-67650686559https://repository.li.mahidol.ac.th/handle/20.500.14594/27649NC37 cells containing the Epstein-Barr virus (EBV) genome do not express the viral glycoprotein-350 (gp350) on the cell surface. Despite being a cancer cell line, NC37 cells show resistance to natural killer (NK) cell cytotoxicity by the standard chromium (51Cr) release assay (CRA). EBV-gp350 has been identified as a ligand for antibody dependent cell-mediated cytotoxicity (ADCC). The stable expression of gp350 on the NC37 cell surface membrane could make this cell line a suitable target for measuring ADCC antibody. The pcDNA3.1-gp350 was transfected into the stably expressing enhanced green fluorescent protein (EGFP)-NC37 cell line. The transfected cells were then selected for expression of gp350 on the cell surface using immunomagnetic bead-based sorting. The gp350-EGFP-NC37 cell line was then re-examined for resistance to NK cytotoxicity, and compared with the standard K562 and EGFP-K562 cell lines using the CRA and a flow cytometric method, respectively. Surprisingly, the gp350-EGFP-NC37 cells, like the parental NC37 cell line, showed comparable resistance to NK cell-mediated cytotoxic activity by the CRA, while demonstrating susceptibility to NK cell cytotoxicity comparable to EGFP expressing K562 cells by the flow cytometric method. The susceptibility of gp350-EGFP-NC37 cells to NK cell cytotoxic activity is dependent on the type of assay. © 2009 Elsevier B.V. All rights reserved.Mahidol UniversityImmunology and MicrobiologyArticleSCOPUS10.1016/j.jviromet.2009.06.009