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Functional analysis and molecular characterization of spontaneously outgrown human lymphoblastoid cell lines

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Abstract

In vitro, the infection of human B-cells with the lymphotropic gammaherpesvirus Epstein-Barr virus (EBV) induces formation of permanently growing lymphoblastoid cell lines (LCL). In a spontaneously outgrown LCL (cell line CSIII), we detected nucleotide sequence variations of the EBV nuclear antigen 1 (EBNA1) RNA that was different from the reference sequence of EBNA1 in the prototypic EBV strain B95-8. In the present study, we molecularly and functionally characterized this virus isolate in comparison to LCL with the prototypic nucleotide sequence. Although we detected high functional similarity between CSIII and the other LCL, our data suggest that the lytic cycle might be ineffective in the CSIII LCL. DNA microarray analysis indicated that RNA binding motif, single stranded interacting protein 1 (RBMS1), which is typically expressed in latency III of EBV to prevent the lytic cycle, was the most overexpressed gene in CSIII LCL.

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Abbreviations

APC:

Antigen presenting cells

BIN1:

Bridging integrator 1

EBER1:

EBV encoded non-coding RNA 1

EBNA1:

EBV nuclear antigen 1

EBV:

Epstein-Barr virus

FACS:

Fluorescence activated cell scanning

FCS:

Fetal calf serum

FITC:

Fluorescein isothiocyanate

LCL:

Lymphoblastoid cell line

LMP2:

Latent membrane protein 2

MFI:

Mean fluorescence intensity

MLC:

Mixed lymphocyte culture

PBMC:

Peripheral blood mononuclear cells

PBS:

Phosphate buffered saline

PCGF2:

Polycomb group ring finger 2

PDCD1LG2:

Programmed cell death 1 ligand 2

PE:

Phycoerythrin

RBMS1:

RNA binding motif, single stranded interacting protein 1

SI:

Stimulation index

TPA:

12-O-tetradecanoylphorbol-13-acetate

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Authors and Affiliations

  1. Department of Pediatrics, Children’s Cancer Research Centre, Martin Luther University Halle-Wittenberg, Ernst-Grube Str. 40, D-06097, Halle, Germany

    Toralf Bernig, Nicole Richter, Ines Volkmer & Martin S. Staege

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  1. Toralf Bernig
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  2. Nicole Richter
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  3. Ines Volkmer
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  4. Martin S. Staege
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Corresponding author

Correspondence to Martin S. Staege.

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Toralf Bernig and Nicole Richter have contributed equally to this work.

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Bernig, T., Richter, N., Volkmer, I. et al. Functional analysis and molecular characterization of spontaneously outgrown human lymphoblastoid cell lines. Mol Biol Rep 41, 6995–7007 (2014). https://doi.org/10.1007/s11033-014-3587-6

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