Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines

Hui-Yuen, Joyce; McAllister, Shane; Koganti, Siva; Hill, Erik; Bhaduri-McIntosh, Sumita

The considerable length of time from EBV infection to establishment of cell lines drives the requirement for quicker and more reliable methods for EBV-driven B cell growth transformation. Using a combination of high titer EBV and an immunosuppressive agent, we are able to consistently infect, transform, and generate LCL from B cells in peripheral blood. This method uses a small amount of peripheral blood mononuclear cells that are infected in vitroclusters of cells can be demonstrated. The presence of CD23 with EBV in the presence of FK506, a T cell immunosuppressant. Traditionally, outgrowth of proliferating B cells is monitored by visualization of microscopic clusters of cells about a week after infection with EBV. Clumps of LCL can be seen by the naked eye after several weeks. We describe an assay to determine early if EBVmediated growth transformation is successful even before microscopic clusters of cells can be demonstrated. The presence of CD23 hi CD58 + cells observed as early as three days post-infection indicates a successful outcome.