Isolation and Culture of EBV-associated Cells

Cell lines include EBV-positive Raji cells (ATCC CCL-86) [59 (link)], EBV-negative Elijah B cells (kindly provided by Prof. A.B. Rickinson), 293 cells (ATCC CRL-1573) [60 (link)], T cells specific for EBNA1 3E10, EBNA3C 5H11, gp350 1D6 and BNRF1 VSD epitopes (kindly provided by Prof. J. Mautner) and autologous LCLs (kindly provided by Prof. J. Mautner). Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque Plus and primary B cells were isolated using Dynabeads CD19 Pan B (Invitrogen) and DETACHaBEAD CD19 kit (Invitrogen). RPMI containing 10% fetal calf serum (F7524, Sigma) was used to culture 293, Raji and Elijah cells. T-cell clones and lines were cultured as previously described [17 (link),35 (link)].

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van Zyl D.G., Tsai M.H., Shumilov A., Schneidt V., Poirey R., Schlehe B., Fluhr H., Mautner J, & Delecluse H.J. (2018). Immunogenic particles with a broad antigenic spectrum stimulate cytolytic T cells and offer increased protection against EBV infection ex vivo and in mice. PLoS Pathogens, 14(12), e1007464.

Publication 2018

Ficoll-Paque Plus Dynabeads CD19 Pan B DETACHaBEAD CD19 kit fetal calf serum

B cells Cell lines Cells Clones Ebna1 Epitopes Fetal calf serum Ficoll Lcls Peripheral blood mononuclear cells T cell

Corresponding Organization : German Center for Infection Research

Other organizations : Frauenklinik Heidelberg, University Hospital Heidelberg, Helmholtz Zentrum München, Technical University of Munich

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Variable analysis

independent variables

Cell lines used: EBV-positive Raji cells (ATCC CCL-86), EBV-negative Elijah B cells, 293 cells (ATCC CRL-1573), T cells specific for EBNA1 3E10, EBNA3C 5H11, gp350 1D6 and BNRF1 VSD epitopes, and autologous LCLs

dependent variables

Not explicitly mentioned

control variables

RPMI containing 10% fetal calf serum (F7524, Sigma) used to culture 293, Raji and Elijah cells
T-cell clones and lines cultured as previously described [17,35]

positive controls

T cells specific for EBNA1 3E10, EBNA3C 5H11, gp350 1D6 and BNRF1 VSD epitopes

negative controls

EBV-negative Elijah B cells

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