874. Long-Term Transgene Expression In Vivo from a Helper Dependent Adenovirus—Epstein-Barr Virus Hybrid Vector

ABSTRACT Epstein-Barr virus (EBV) episomes are stably maintained in permissive proliferating cell lines due to EBV nuclear antigen 1 (EBNA-1) protein-mediated replication and segregation. Previous studies showed the ability of EBV episomes to confer long-term transgene expression and correct genetic defects in deficient cells. To achieve quantitative delivery of EBV episomes in vitro and in vivo, we developed a binary helper-dependent adenovirus (HDA)-EBV hybrid system that consists of one HDA vector for the expression of Cre recombinase and a second HDA vector that contains all of the sequences for the EBV episome flanked by loxP sites. Upon coinfection of cells, Cre expressed from the first vector recombined loxP sites on the second vector. The resulting circular EBV episomes expressed a transgene and contained the EBV-derived family of repeats, an EBNA-1 expression cassette, and 19 kb of human DNA that functions as a replication origin in mammalian cells. This HDA-EBV hybrid system transformed 40% of cultured cells. Transgene expression in proliferating cells was observed for over 20 weeks under conditions that selected for the expression of the transgene. In the absence of selection, EBV episomes were lost at a rate of 8 to 10% per cell division. Successful delivery of EBV episomes in vivo was demonstrated in the liver of transgenic mice expressing Cre from the albumin promoter. This novel gene transfer system has the potential to confer long-term episomal transgene expression and therefore to correct genetic defects with reduced vector-related toxicity and without insertional mutagenesis.

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Epstein–Barr virus-based plasmid enables inheritable transgene expression in mouse cerebral cortex

PLoS ONE ◽

10.1371/journal.pone.0258026 ◽

2021 ◽

Vol 16 (9) ◽

pp. e0258026

Author(s):

Tomoko Satake

Keyword(s):

Cerebral Cortex ◽

Glial Cells ◽

Transgene Expression ◽

Epstein Barr Virus ◽

Continuous Development ◽

Shrna Expression ◽

Embryonic Mouse ◽

Barr Virus ◽

Epstein Barr

Continuous development of the cerebral cortex from the prenatal to postnatal period depends on neurons and glial cells, both of which are generated from neural progenitor cells (NPCs). Owing to technical limitations regarding the transfer of genes into mouse brain, the mechanisms behind the long-term development of the cerebral cortex have not been well studied. Plasmid transfection into NPCs in embryonic mouse brains by in utero electroporation (IUE) is a widely used technique aimed at expressing transgenes in NPCs and their recent progeny neurons. Because the plasmids in NPCs are attenuated with each cell division, the transgene is not expressed in their descendants, including glial cells. The present study shows that an Epstein–Barr virus-based plasmid (EB-oriP plasmid) is helpful for studying long-term cerebral cortex development. The use of the EB-oriP plasmid for IUE allowed transgene expression even in the descendant progeny cells of adult mouse brains. Combining the EB-oriP plasmid with the shRNA expression cassette allowed examination of the genes of interest in the continuous development of the cerebral cortex. Furthermore, preferential transgene expression was achieved in combination with cell type-specific promoter-driven transgene expression. Meanwhile, introducing the EB-oriP plasmid twice into the same individual embryos during separate embryonic development stages suggested heterogeneity of NPCs. In summary, IUE using the EB-oriP plasmid is a novel option to study the long-term development of the cerebral cortex in mice.

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Sustained transgene expression in vitro and in vivo using an Epstein–Barr virus replicon vector system combined with HVJ liposomes

Gene Therapy ◽

10.1038/sj.gt.3300711 ◽

1998 ◽

Vol 5 (8) ◽

pp. 1031-1037 ◽

Cited By ~ 61

Author(s):

Y Saeki ◽

M Wataya-Kaneda ◽

K Tanaka ◽

Y Kaneda

Keyword(s):

Transgene Expression ◽

Epstein Barr Virus ◽

Vector System ◽

Barr Virus ◽

Replicon Vector ◽

Epstein Barr

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14. High Efficiency and Long-Term Persistence In Vivo from a Helper Dependent Adenovirus/Epstein-Barr Virus Hybrid Vector

Molecular Therapy ◽

10.1016/j.ymthe.2006.08.025 ◽

2006 ◽

Vol 13 ◽

pp. S6

Author(s):

Sean D. Gallaher ◽

Jose S. Gil ◽

Arnold J. Berk

Keyword(s):

Epstein Barr Virus ◽

High Efficiency ◽

Barr Virus ◽

Epstein Barr

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825. In Vivo Gene Delivery and Long-Term Expression in Mouse by a Helper Dependent Adenovirus-Epstein-Barr Virus Hybrid Vector

Molecular Therapy ◽

10.1016/s1525-0016(16)40228-5 ◽

2008 ◽

Vol 16 ◽

pp. S308

Keyword(s):

Gene Delivery ◽

Epstein Barr Virus ◽

Barr Virus ◽

In Vivo Gene Delivery ◽

Epstein Barr

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Long-term eradication of extranodal natural killer/T-cell lymphoma, nasal type, by induced pluripotent stem cell-derived Epstein-Barr virus-specific rejuvenated T cells in vivo

Haematologica ◽

10.3324/haematol.2019.223511 ◽

2019 ◽

Vol 105 (3) ◽

pp. 796-807 ◽

Cited By ~ 3

Author(s):

Miki Ando ◽

Jun Ando ◽

Satoshi Yamazaki ◽

Midori Ishii ◽

Yumi Sakiyama ◽

...

Keyword(s):

Epstein Barr Virus ◽

Cell Lymphoma ◽

Induced Pluripotent Stem Cell ◽

Natural Killer T Cell ◽

Barr Virus ◽

Epstein Barr ◽

Induced Pluripotent ◽

Killer T Cell

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Epstein-Barr virus–specific human T lymphocytes expressing antitumor chimeric T-cell receptors: potential for improved immunotherapy

Blood ◽

10.1182/blood.v99.6.2009 ◽

2002 ◽

Vol 99 (6) ◽

pp. 2009-2016 ◽

Cited By ~ 145

Author(s):

Claudia Rossig ◽

Catherine M. Bollard ◽

Jed G. Nuchtern ◽

Cliona M. Rooney ◽

Malcolm K. Brenner

Keyword(s):

T Cells ◽

T Cell ◽

Epstein Barr Virus ◽

Chimeric Receptor ◽

Chimeric Receptors ◽

Barr Virus ◽

Epstein Barr ◽

And Function

Abstract Primary T cells expressing chimeric receptors specific for tumor or viral antigens have considerable therapeutic potential. Unfortunately, their clinical value is limited by their rapid loss of function and failure to expand in vivo, presumably due to the lack of costimulator molecules on tumor cells and the inherent limitations of signaling exclusively through the chimeric receptor. Epstein-Barr virus (EBV) infection of B lymphocytes is near universal in humans and stimulates high levels of EBV-specific helper and cytotoxic T cells, which persist indefinitely. Our clinical studies have shown that EBV-specific T cells generated in vitro will expand, persist, and function for more than 6 years in vivo. We now report that EBV-specific (but not primary) T cells transduced with tumor-specific chimeric receptor genes can be expanded and maintained long-term in the presence of EBV-infected B cells. They recognize EBV-infected targets through their conventional T-cell receptor and tumor targets through their chimeric receptors. They efficiently lyse both. EBV-specific T cells expressing chimeric antitumor receptors may represent a new source of effector cells that would persist and function long-term after their transfer to cancer patients.

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EBV-LMP1 promotes radioresistance by inducing protective autophagy through BNIP3 in nasopharyngeal carcinoma

Cell Death and Disease ◽

10.1038/s41419-021-03639-2 ◽

2021 ◽

Vol 12 (4) ◽

Author(s):

San Xu ◽

Zhuan Zhou ◽

Xingzhi Peng ◽

Xuxiu Tao ◽

Peijun Zhou ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Crucial Role ◽

Epstein Barr Virus ◽

Interacting Protein ◽

Multiple Tumors ◽

Barr Virus ◽

Signaling Axis ◽

Epstein Barr ◽

Adenovirus E1b

AbstractStudies have indicated that dysfunction of autophagy is involved in the initiation and progression of multiple tumors and their chemoradiotherapy. Epstein–Barr virus (EBV) is a lymphotropic human gamma herpes virus that has been implicated in the pathogenesis of nasopharyngeal carcinoma (NPC). EBV encoded latent membrane protein1 (LMP1) exhibits the properties of a classical oncoprotein. In previous studies, we experimentally demonstrated that LMP1 could increase the radioresistance of NPC. However, how LMP1 contributes to the radioresistance in NPC is still not clear. In the present study, we found that LMP1 could enhance autophagy by upregulating the expression of BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3). Knockdown of BNIP3 could increase the apoptosis and decrease the radioresistance mediated by protective autophagy in LMP1-positive NPC cells. The data showed that increased BNIP3 expression is mediated by LMP1 through the ERK/HIF1α signaling axis, and LMP1 promotes the binding of BNIP3 to Beclin1 and competitively reduces the binding of Bcl-2 to Beclin1, thus upregulating autophagy. Furthermore, knockdown of BNIP3 can reduce the radioresistance promoted by protective autophagy in vivo. These data clearly indicated that, through BNIP3, LMP1 induced autophagy, which has a crucial role in the protection of LMP1-positive NPC cells against irradiation. It provides a new basis and potential target for elucidating LMP1-mediated radioresistance.

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