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Updated: Apr 13, 2026
Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines
Published on: November 8, 2011
Epstein-Barr virus genetic variation in lymphoblastoid cell lines derived from Kenyan pediatric population
Kenneth O Simbiri1, Nicholas A Smith1, Richard Otieno2
1Department of Microbiology and Immunology, Upstate Medical University, Syracuse, New York, United States of America.
Insights
Genetic variations of Epstein-Barr virus (EBV) strains in Kenya were analyzed using next-generation sequencing. Different EBV types, including EBV-1 and EBV-2, were found to co-exist within the same geographic region.
Area of Science:
- Virology
- Genetics
- Molecular Biology
Background:
- Epstein-Barr virus (EBV) is linked to Burkitt's lymphoma (BL).
- Sub-Saharan Africa has endemic BL and harbors both EBV Type 1 (EBV-1) and EBV Type 2 (EBV-2) strains.
- Limited knowledge exists regarding EBV strain genetic diversity in sub-Saharan Africa.
Purpose of the Study:
- To investigate the genetic variation of Epstein-Barr virus (EBV) strains in Kenya.
- To analyze inter- and intra-strain variations within EBV genomes from a specific geographic region.
- To assess the utility of next-generation sequencing (NGS) for detailed EBV genome analysis.
Main Methods:
- Generation of spontaneous lymphoblastoid cell lines (LCLs) from Kenyan samples.
- Polymerase chain reaction (PCR) amplification of the EBV genome.
- Next-generation sequencing (NGS) for genome sequencing.
- Phylogenetic and single nucleotide polymorphism (SNP) analyses.
Main Results:
- Phylogenetic analysis revealed EBV strains related to both EBV-1 and EBV-2.
- One sample (LCL10) showed closer relation to EBV-2, while three LCLs were closer to EBV-1.
- SNP analysis indicated clustering of LCL variants.
- Latent genes (EBNA-1, BLLF1, BPLF1, BRRF2) were found to be less conserved than lytic genes.
Conclusions:
- Next-generation sequencing (NGS) is effective for detailed analysis of EBV genome variations.
- Different genetic variations of EBV can coexist within a single geographic area.
- Further research is needed to understand the implications of co-existing EBV variants.
- Findings contribute to understanding pathogenic EBV variants in EBV-associated malignancies.
Abstract:
Epstein-Barr virus (EBV) is associated with Burkitt's lymphoma (BL), and in regions of sub-Saharan Africa where endemic BL is common, both the EBV Type 1 (EBV-1) and EBV Type 2 strains (EBV-2) are found. Little is known about genetic variation of EBV strains in areas of sub-Saharan Africa. In the present study, spontaneous lymphoblastoid cell lines (LCLs) were generated from samples obtained from Kenya. Polymerase chain reaction (PCR) amplification of the EBV genome was done using multiple primers and sequenced by next-generation sequencing (NGS). Phylogenetic analyses against the published EBV-1 and EBV-2 strains indicated that one sample, LCL10 was closely related to EBV-2, while the remaining 3 LCL samples were more closely related to EBV-1. Moreover, single nucleotide polymorphism (SNP) analyses showed clustering of LCL variants. We further show by analysis of EBNA-1, BLLF1, BPLF1, and BRRF2 that latent genes are less conserved than lytic genes in these LCLs from a single geographic region. In this study we have shown that NGS is highly useful for deciphering detailed inter and intra-variations in EBV genomes and that within a geographic region different EBV genetic variations can co-exist, the implications of which warrant further investigation. The findings will enhance our understanding of potential pathogenic variants critical to the development and maintenance of EBV-associated malignancies.
