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Non‐integrating lentiviral vectors for specific killing of Epstein–Barr virus nuclear antigen 1‐positive B cell lymphoma cells
Author(s) -
Flynn Ryan P.,
Zacharias Jeana,
Zhou Xianzheng,
Can Mark L.,
Philpott Nicola J.
Publication year - 2011
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1601
Subject(s) - biology , thymidine kinase , transgene , epstein–barr virus , virology , viral vector , antigen , herpes simplex virus , b cell , population , virus , lymphoma , microbiology and biotechnology , cell culture , bystander effect , gene , antibody , immunology , recombinant dna , genetics , medicine , environmental health
Background Epstein–Barr virus (EBV) causes a range of life‐threatening B‐lymphocyte malignancies but, despite the use of various strategies, treatment remains problematic. Methods In the present study, we developed a non‐integrating lentiviral vector (NILV) that mediates specific killing of EBV nuclear antigen 1 (EBNA1)‐expressing cells with minimal toxicity to EBNA1‐negative cells. The EBV family of repeats (FR) was cloned intok the NILV genome upstream of various transgenes. Results The presence of the FR in the NILV genome induced transcriptional up‐regulation and prolonged the expression of a transgene specifically in EBNA1‐positive B cells. Transgene expression from an FR‐containing NILV was also prolonged in EBV‐transformed cells compared to an FR‐negative NILV. We found that the delivery of an FR‐containing NILV encoding herpes simplex virus 1 thymidine kinase (TK) lead to the killing of more than 99% of EBNA1‐positive B cells with minimal toxicity to EBNA1‐negative cells in the presence of gancyclovir. EBNA1‐positive cells were not killed by an FR‐negative vector containing the TK gene. An FR‐TK‐containing NILV also specifically killed EBNA1‐containing cells in a mixed population of EBNA1‐positive and EBNA1‐negative cells, thus confirming that NILV‐FR‐TK‐mediated killing is specific for EBNA1‐expressing cells. Conclusions Transgene expression from our NILVs is both EBNA1‐specific and dependent upon the presence of the FR. The results obtained in the present study indicate that NILVs have potential use in the treatment of EBV‐associated B cell malignancies. Copyright © 2011 John Wiley & Sons, Ltd.