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Virus and cell RNAs expressed during Epstein-Barr virus replication
Jing Yuan1, Ellen Cahir-McFarland, Bo Zhao
1Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Channing Laboratory, Boston, MA 02115, USA.
Journal of Virology
|February 14, 2006
Summary
Epstein-Barr virus (EBV) replication in Akata Burkitt B lymphoblasts was induced by immunoglobulin G (IgG) cross-linking. This process led to increased EBV replication, altered cellular RNA levels, and late expression of latency III (LTIII) RNAs and proteins.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Epstein-Barr virus (EBV) establishes latency in B cells.
- Understanding EBV replication dynamics is crucial for disease pathogenesis.
- Akata Burkitt B lymphoblasts are a model system for studying EBV latency and replication.
Purpose of the Study:
- To investigate the changes in EBV and host cell RNA levels during EBV replication.
- To characterize the temporal expression of EBV RNAs, including latency III (LTIII) transcripts.
- To determine the impact of EBV replication on cellular gene expression.
Main Methods:
- Induction of EBV replication in latency 1-infected Akata Burkitt B lymphoblasts using immunoglobulin G (IgG) cross-linking.
- Assay of EBV replication via membrane gp350 expression.
- Quantification of EBV and cellular RNA levels using hybridization techniques.
- Analysis of protein and viral DNA synthesis inhibitors' effects.
- Propidium iodide exclusion assay to assess cell viability.
Main Results:
- IgG cross-linking significantly increased EBV replication, with gp350 expression rising from 5% to over 50% within 48 hours.
- Temporal changes in EBV RNA levels (BZLF1, BMLF1, BRLF1) followed expected patterns of viral gene expression.
- Surprisingly, latency III (LTIII) RNAs (LMP1, LMP2, EBNA2, etc.) were detected and peaked late in replication (24-48 hours).
- EBV replication altered levels of 401 Akata cell RNAs, with 122 changing twofold or more, including mitogen-activated protein kinase pathways.
- Late LTIII expression correlated with the induction of NF-kappaB responsive genes (IkappaBalpha, A20).
Conclusions:
- EBV replication, induced by IgG cross-linking, leads to significant changes in both viral and cellular RNA expression.
- The late expression of LTIII RNAs and proteins is a characteristic feature of EBV replication in B lymphoblasts, mirroring observations in immune-compromised individuals.
- EBV replication impacts host cell gene expression, affecting pathways like mitogen-activated protein kinase and NF-kappaB signaling, indicating vital cell function during late replication.