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题目:: Stable expression of EBV-gp350 on the surface of NC37 cells confers natural killer (NK)-cell susceptibility or resistance, depending on the assay used to assess NK-mediated function.
作者:: Sumonwiriya(Manutsanun),Horhthongkham(Navin),Pattanapanyasat(Kovit),Ampol(Silawun),Sutthent(Reungpung),Kantakamalakul(Wannee)
状态:: 发布时间2009-07-27 , 更新时间 2009-07-27
期刊:: J Virol Methods
摘要:: NC37 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 ((51)Cr) 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.
语言:: eng
DOI:: 10.1016/j.jviromet.2009.06.009

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