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The AKATA cell line, derived from Burkitt’s lymphoma, is a widely used model to study Epstein-Barr virus (EBV) latency and reactivation. EBV is a ubiquitous herpesvirus linked to a range of cancers, including Burkitt lymphoma, and typically establishes a latent infection within B cells. In AKATA cells, EBV is maintained in an episomal state with a type I latency program, expressing a limited set of viral genes such as EBNA-1, EBER RNAs, and BamHI-A rightward transcripts (BARTs). This restricted gene expression allows the virus to persist in the host without initiating a full lytic cycle. However, AKATA cells can be triggered to enter the lytic phase, where the virus actively replicates and produces progeny. This reactivation is commonly induced through cross-linking surface immunoglobulins, which makes AKATA cells an excellent tool for studying EBV reactivation dynamics and viral gene regulation.
Research utilizing the AKATA cell line has also examined the impact of chemotherapeutic agents on EBV reactivation. For instance, drugs like etoposide and doxorubicin have been shown to influence viral latency. Etoposide induces apoptosis in AKATA cells but reactivates EBV less effectively than doxorubicin, which promotes higher levels of lytic gene expression and viral progeny production. Additionally, studies involving gene editing techniques, such as CRISPR/Cas9, have explored the role of epigenetic regulators in AKATA cells. For example, knockout of the histone methyltransferase EZH2 in AKATA cells disrupts the maintenance of latency by reducing the trimethylation of histone H3K27, leading to increased expression of both latent and lytic EBV genes, as well as enhanced viral replication and cell proliferation.
AKATA cells also display distinct phenotypic characteristics based on EBV presence, such as increased sensitivity to apoptosis-inducing agents and variations in gene expression related to apoptotic pathways. These differences make EBV-positive AKATA cells a powerful model for dissecting EBV's influence on host cell survival, gene expression, and the virus's lifecycle, particularly in the context of cancer development and potential therapeutic interventions targeting EBV-associated malignancies.
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Viruses: Transformant: EBV
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- subculturing: Gather the suspension cells in a 15 ml tube and gently wash the adherent cells with PBS lacking calcium and magnesium (use 3-5 ml for T25 flasks and 5-10 ml for T75 flasks). Apply Accutase (1-2 ml for T25 flasks, 2.5 ml for T75 flasks) ensuring full coverage of the cell layer. Allow the cells to incubate at room temperature for 10 minutes. Following incubation, combine and centrifuge both the suspension and adherent cells. After centrifugation, carefully resuspend the cell pellet and transfer the cell suspension into new flasks containing fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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