Efficient gene transfer into lymphoma cells using adenoviral vectors combined with lipofection

Buttgereit, Peter; Weineck, Silvia; Röpke, Gabriele; Märten, Angela; Brand, Karsten; Heinicke, Thomas; Caselmann, Wolfgang H.; Huhn, D.; Schmidt‐Wolf, Ingo G.H.

doi:10.1038/sj.cgt.7700209

Cited by 31 publications

(32 citation statements)

References 28 publications

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“…Although we have not directly addressed the role of this integrin in our studies, we note that several of the cell lines in which we successfully grew WNV in this study, including the K562, Raji, and CHO-K1 cell lines, previously have been shown to lack ␣v␤3 integrin expression (14,61). Thus, ␣v␤3 is not absolutely required for WNV infection.…”

Section: Discussionmentioning

confidence: 92%

West Nile Virus Discriminates between DC-SIGN and DC-SIGNR for Cellular Attachment and Infection

Davis

Nguyen

Hanna

et al. 2006

J Virol

306

307

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The C-type lectins DC-SIGN and DC-SIGNR bind mannose-rich glycans with high affinity. In vitro, cells expressing these attachment factors efficiently capture, and are infected by, a diverse array of appropriately glycosylated pathogens, including dengue virus. In this study, we investigated whether these lectins could enhance cellular infection by West Nile virus (WNV), a mosquito-borne flavivirus related to dengue virus. We discovered that DC-SIGNR promoted WNV infection much more efficiently than did DC-SIGN, particularly when the virus was grown in human cell types. The presence of a single N-linked glycosylation site on either the prM or E glycoprotein of WNV was sufficient to allow DC-SIGNR-mediated infection, demonstrating that uncleaved prM protein present on a flavivirus virion can influence viral tropism under certain circumstances. Preferential utilization of DC-SIGNR was a specific property conferred by the WNV envelope glycoproteins. Chimeras between DC-SIGN and DC-SIGNR demonstrated that the ability of DC-SIGNR to promote WNV infection maps to its carbohydrate recognition domain. WNV virions and subviral particles bound to DC-SIGNR with much greater affinity than DC-SIGN. We believe this is the first report of a pathogen interacting more efficiently with DC-SIGNR than with DC-SIGN. Our results should lead to the discovery of new mechanisms by which these well-studied lectins discriminate among ligands.The first step in viral entry is the stable attachment of the virion to the surface of a new target cell, a process that can be inefficient for many viruses (16,34,62,73). Cellular proteins that facilitate productive infection by increasing the efficiency of virus binding, but whose presence is not absolutely required for viral entry, are often referred to as attachment factors (4). Two of the most extensively studied attachment factors are the lectins DC-SIGN (CD209) (18, 29) and DC-SIGNR (L-SIGN) (CD209L) (7,67,78). Both are tetrameric type II transmembrane proteins containing calcium-dependent (C-type) carbohydrate recognition domains (CRDs) (55). DC-SIGN is highly expressed in monocyte-derived dendritic cells (MDDCs) in vitro (29) and at lower levels (86) in vivo in subsets of macrophages (45,53,79) and dendritic cells (23,29,40,80,86). DC-SIGNR is expressed on microvascular endothelial cells, especially in the liver sinusoids and lymph nodes (7,67,81). By facilitating virion attachment, DC-SIGN and DC-SIGNR [henceforth referred to collectively as DC-SIGN(R)] can greatly increase the susceptibility of permissive cells to infection by a wide array of enveloped viruses or allow nonpermissive cells to capture and transmit these viruses to target cells in trans (3,17,35,47,52,60,76,84).Viruses that bind to DC-SIGN(R) appear to do so via highmannose, N-linked glycans on their glycoproteins (44,48,51). This fact is readily explained by crystallographic studies demonstrating that mannose-rich oligosaccharides fit into elongated binding sites in the CRDs of DC-SIGN(R) (24). In addition to recognizing viral...

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Section: Discussionmentioning

confidence: 92%

West Nile Virus Discriminates between DC-SIGN and DC-SIGNR for Cellular Attachment and Infection

Davis

Nguyen

Hanna

et al. 2006

J Virol

306

307

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“…14,15,33 Our results showed that the transfection efficiencies achieved with the cell lines used were high, which makes it possible to successfully transfer the p53 gene without the toxicity associated with high virus concentrations. 16 We observed high expression of CAR on the surface of the lymphoma cell lines Raji and Daudi. The cell lines LAM53, CA46, and BL41 showed a lower expression of CAR and lower transfection efficiencies, which suggests that the adenoviral entry into these cells is mediated by CAR.…”

Section: Discussionmentioning

confidence: 76%

“…14,15 However, we previously demonstrated very efficient gene transfer in BL cell lines and primary lymphoma cells after transfection with adenoviral vectors. 16 Adenoviral gene delivery to specific cell types seems to be limited by the availability of the coxsackie adenoviral receptor ( CAR ) and integrins alpha v beta. 17 ± 20 We showed a high expression of CAR on the surface of some lymphoma cells, suggesting that the adenoviral entry into these cells is mediated by CAR.…”

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confidence: 99%

Effects of adenoviral wild-type p53 gene transfer in p53 -mutated lymphoma cells

et al. 2001

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The present study assessed the role of adenoviral vector ± mediated wild -type p53 gene transfer in B lymphoma cells. Deficiency of p53 -mediated cell death is common in human cancer contributing to both tumorigenesis and chemoresistance. Lymphoma cells are being considered as suitable targets for gene therapy protocols. Recently, we reported an adenoviral protocol leading to highly efficient gene transfer to B lymphoma cells. All lymphoma cell lines ( n = 5 ) tested here showed mutations in the p53 gene locus. The aim of this work was to transduce lymphoma cells with the wild -type p53 gene. Using this protocol, 88% of Raji, 75% of Daudi, and 45% of OCI -Ly8 -LAM53 cells were transfected with the reporter gene green fluorescent protein at a multiplicity of infection of 200. The expression of green fluorescent protein in CA46 and BL41 cells was 27% and 42%, respectively. At this multiplicity of infection, growth characteristics of lymphoma cell lines were not changed significantly. In contrast, cells transduced with wild -type p53 gene showed an inhibition of proliferation as well as an increase in apoptosis. Cell loss by apoptosis after p53 gene transfer was up to 40% as compared to transduction with an irrelevant vector. In addition, we determined the effects of DNA damage produced by the DNA topoisomerase II inhibitor etoposide on wild -type p53 transfected lymphoma cells. In Ad -p53 ± transfected Raji cells, treatment with the drug resulted in a marked increase of cell loss in comparison to Ad --Gal ± transfected cells ( 45% vs. 77% ). Interestingly, performing cytotoxicity studies, we could show an increased sensitivity of Raji and Daudi cells against immunological effector cells. In conclusion, transduction of wild -type p53 into lymphoma cells expressing mutated p53 was efficient and led to inhibition of proliferation and increase in apoptotic rate in some cell lines dependent on p53 mutation. This protocol should have an impact on the use of lymphoma cells in cancer gene therapy protocols.

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“…20 While lymphoid cells would appear ideal for testing the Ad-ADH system, the routine introduction of simple Ad5 constructs into lymphoid cells is currently difficult to achieve. 30 In Figure 3a and b, the effects of the enhanced ethanol metabolism and acetaldehyde production on the growth and viability of wild type and Ad-ADH-transduced CMT-64 cells is shown. Native CMT-64 cells are unaffected by ethanol exposure while transduction with Ad-ADH leads to slowing of cell division and cell death, which was enhanced with either higher ratios of virus or longer exposure to ethanol.…”

Section: Discussionmentioning

confidence: 99%

Suicide gene therapy: conversion of ethanol to acetaldehyde mediated by human beta 2 alcohol dehydrogenase

et al. 2004

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Acetaldehyde (AcH) produced in the physiological metabolism of ethanol can be potentially toxic and immunomodulating. The antitumour activity of a suicide gene system using adenovirus delivered alcohol dehydrogenase (ADH) to convert ethanol to acetaldehyde inside cancer cells has been investigated in vitro and in vivo. In vitro experiments confirmed the toxicity of acetaldehyde to a number of tumour cell lines. Daudi lymphoma cells grown in normal media increased by Day 4 to 650% of their starting number, while those exposed to 250 mM, 500 mM and 1 mM acetaldehyde reached 138, 30 and 5% respectively. Adenocarcinoma cells appeared to be less sensitive with CMT-64 cells and HeLa cells numbering 105 and 53% of their starting number by Day 4 with 1 mM acetaldehyde. After transduction with an adenovirus containing the human ADH beta 2 cDNA, CMT-64 cells exposed to 20 mM ethanol had a reduction in number to 74% by Day 2 and to 36% by Day 4. In a preclinical model with Ad-ADH CMT-64 cells, mice exposed to daily pulses of ethanol for 5 days formed tumours only 30% on Day 6 and 42% on Day 13 of the volume of those in mice exposed to water.The ability of this easily administered suicide gene system to produce significant effects on cell proliferation in vivo suggests that further optimized development is warranted.

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Efficient gene transfer into lymphoma cells using adenoviral vectors combined with lipofection

Cited by 31 publications

References 28 publications

West Nile Virus Discriminates between DC-SIGN and DC-SIGNR for Cellular Attachment and Infection

West Nile Virus Discriminates between DC-SIGN and DC-SIGNR for Cellular Attachment and Infection

Effects of adenoviral wild-type p53 gene transfer in p53 -mutated lymphoma cells

Suicide gene therapy: conversion of ethanol to acetaldehyde mediated by human beta 2 alcohol dehydrogenase

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