CD19 signalling improves the Epstein–Barr virus‐induced immortalization of human B cell

Abstract

Epstein-Barr virus (EBV) infection in vitro immortalizes primary B cells and generates B lymphoblastoid cell lines (LCLs). These EBV-LCLs have been used for several purposes in immunological and genetic studies, but some trials involving these transformations fail for unknown reasons, and several EBV-LCLs do not grow in normal culture. In this study, we improved the immortalization method by CD19 and B-cell receptor (BCR) co-ligation. This method shortens the time required for the immortalization and generation of EBV-LCLs but does not alter the cell phenotype of the LCLs nor the expression of the EBV genes. In particular, the CD19 and BCR co-ligation method was found to be the most effective method examined. EBV-infected B cells induced by CD19 and/or BCR ligation expressed the intracellular latent membrane protein LMP-1 earlier than EBV-infected B cells, and the expression of intracellular LMP-1 was found to be closely related to the time of immortalization. These results suggest that the modified method, using CD19 and/or BCR ligation, may efficiently generate EBV-LCLs, by expressing intracellular LMP-1 at an early stage.

High‐potentiality preliminary selection criteria and transformation time‐dependent factors analysis for establishing Epstein–Barr virus transformed human lymphoblastoid cell lines

“…(2003) found that in the transformation efficiency of a group infected with tetradecanoyl phorbol acetate, the induced EBV was higher than that of their control group. Recently, Hur et al (2005) found that numbers of cell clumps and expression of CD38 in LCLs could be enhanced by CD19 ligation. Mello et al .…”

Mucosal Inducible NO Synthase–Producing IgA+ Plasma Cells in Helicobacter pylori–Infected Patients

“…To ensure sufficient cell numbers for these experiments, we used EBVtransformed B cells as a model to study the expression of CD38 and iNOS, as well as the production of different pro-and antiinflammatory cytokines, including IL-1b, IL-2, IL-6, IL-8, TNF-a, IFN-g, IL-4, and IL-10. According to previous studies, EBV transformation of primary human B cells changes various cellular properties (23,24), such as the expression of the cell activation-associated Ag CD38 (25), and the expression of iNOS (26). Furthermore, B-LCLs have been described as expressing various cytokines with a high degree of heterogeneity (24,(27)(28)(29)(30)(31)(32).…”

Evaluation of EBV transformation of human memory B-cells isolated by FACS and MACS techniques

“…It could be postulated that, due to the attached CD19-conjugated magnetic beads on the B-cells, the positively-selected cells had less capability to undergo EBV transformation compared to negativelyselected counterparts. However, Hur et al (2005) reported that attachment of anti-CD19 polyclonal antibodies to B-cells during MACS-mediated positive selection increased expression of cytosolic (but not membranous) LMP-1 proteins that have a critical role in EBV transformation of B-cells. It is also possible the binding of CD19-beads may have induced conformational changes that disrupted the EBV ability to infect the cells via CD21, as both molecules are part of the B-cell receptor complex.…”

“…For example, CD19 and BCR co-ligation has been shown to boost EBV transformation and establishment of LCL (Hur et al 2005). Nevertheless, little is known about the effect of choice of method by which B-cells are isolated on these parameters.…”