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A new primary effusion lymphoma-derived cell line yields a highly infectious Kaposi's sarcoma herpesvirus-containing
J S Cannon1, D Ciufo, A L Hawkins
1Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Insights
A new primary effusion lymphoma cell line, JSC-1, efficiently produces infectious Kaposi's sarcoma herpesvirus (KSHV). This KSHV efficiently infects endothelial cells, offering a valuable tool for studying KSHV pathogenesis.
Area of Science:
- Oncology
- Virology
- Immunology
Background:
- Primary effusion lymphoma (PEL) is an aggressive non-Hodgkin lymphoma associated with human immunodeficiency virus (HIV) infection.
- Kaposi's sarcoma herpesvirus (KSHV) is the causative agent of PEL.
- Efficient production of infectious KSHV is crucial for studying its biology and developing therapeutic strategies.
Purpose of the Study:
- To establish and characterize a novel PEL cell line for KSHV production.
- To compare the infectious KSHV yield from the new cell line with existing PEL lines.
- To investigate the effects of KSHV infection on human dermal microvascular endothelial cells (DMVECs).
Main Methods:
- Establishment of the JSC-1 PEL cell line from patient ascitic fluid.
- Flow cytometry and Southern blot hybridization for cellular lineage and gene expression analysis.
- Reverse transcription-PCR and immunohistochemistry for viral gene expression.
- Infection of DMVECs with KSHV from different PEL cell lines and quantification of viral DNA.
Main Results:
- JSC-1 cells expressed CD45 and lambda light-chain, with immunoglobulin heavy-chain gene rearrangements indicating B-cell lineage.
- JSC-1 cells exhibited latent Epstein-Barr virus (EBV) expression and both latent and lytic Kaposi's sarcoma herpesvirus (KSHV) gene expression.
- Tetradecanoyl phorbol acetate and butyrate upregulated KSHV lytic gene expression in JSC-1 cells.
- JSC-1 cell supernatant yielded significantly higher infectious KSHV compared to BC-3 and BCP-1 cell lines.
- KSHV infection of DMVECs resulted in morphological changes, LANA expression, and increased mitoses.
Conclusions:
- The JSC-1 cell line is a valuable source of highly infectious KSHV.
- JSC-1 provides a robust model for studying KSHV replication and infection.
- KSHV infection induces specific cellular changes in endothelial cells, relevant to KSHV pathogenesis.
Abstract:
A primary effusion lymphoma (PEL) cell line, JSC-1, that yields highly infectious Kaposi's sarcoma herpesvirus (KSHV) supernatants was established from the ascitic fluid of a human immunodeficiency virus-positive patient. Flow cytometry showed strong expression of CD45 and lambda light-chain restriction. Southern blot hybridization showed immunoglobulin heavy-chain gene rearrangements in the tumor and the resultant cell line consistent with B-cell lineage. Expression of viral genes was assessed by reverse transcription-PCR and immunohistochemistry. Only latent Epstein-Barr virus (EBV) gene expression was detected, and this was at a low level. In contrast, lytic and latent KSHV gene expression were detected. Tetradecanoyl phorbol acetate and butyrate upregulated KSHV lytic expression, but not EBV lytic expression. Viral supernatant from JSC-1 was much more efficient at infecting primary human dermal microvascular endothelial cells (DMVECs) with KSHV than supernatants from BC-3 or BCP-1 PEL cell lines. Quantitation of viral yields produced by the PEL lines showed at least 2 orders of magnitude more DNase I-resistant KSHV DNA in the JSC-1 supernatant compared to BC-3 or BCP-1 supernatants. KSHV infection in DMVECs was associated with a change from a cobblestone to a spindle shape, LANA expression, and an increased number of mitoses.