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Journal of Virology, January 2000, p. 354-362, Vol. 74, No. 1
Department of Immunology, Scripps Research
Institute, La Jolla, California 92037,1 and
Genetic Therapy, Inc., Gaithersburg, Maryland
208782
Received 8 June 1999/Accepted 22 September 1999
While adenovirus (Ad) gene delivery vectors are useful in many gene
therapy applications, their broad tropism means that they cannot be
directed to a specific target cell. There are also a number of cell
types involved in human disease which are not transducible with
standard Ad vectors, such as Epstein-Barr virus (EBV)-transformed B
lymphocytes. Adenovirus binds to host cells via the viral fiber protein, and Ad vectors have previously been retargeted by modifying the fiber gene on the viral chromosome. This requires that the modified
fiber be able to bind to the cell in which the vector is grown, which
prevents truly specific vector targeting. We previously reported a gene
delivery system based on a fiber gene-deleted Ad type 5 (Ad5) vector
(Ad5. Adenovirus (Ad)-based gene delivery
vectors efficiently infect many different cells and tissues, making
them promising tools for gene therapy (15, 23, 24, 52).
However, this broad tropism means that gene delivery cannot be directed
to a specific target cell. For example, a large fraction of
intravenously administered Ad is retained by the liver, which may not
be a desirable target (18, 44). Ad type 5 (Ad5) also
transduces dendritic cells, which present antigens very efficiently
(20, 60) and may exacerbate the antivector immune response.
Vectors with different targeting preferences might eliminate these
problems, allowing use of a lower (and therefore less immunogenic)
total particle dose. Altered vector tropism would also extend the use
of Ad-mediated gene delivery to cell types not infected by Ad5, such as
cells of the hematopoietic system.
Infection by Ad involves two distinct virus-cell interactions (reviewed
in reference 2). Attachment to a target cell occurs via high-affinity binding of the viral fiber protein to a specific cell
surface receptor (6, 39). Internalization is then mediated by interaction of the viral penton base protein with cellular Gene therapy strategies have been proposed for treatment of
Epstein-Barr virus (EBV)-induced diseases (11, 17, 21, 22, 42,
56). EBV is associated with such life-threatening disorders as
transplant-associated lymphoproliferation (30, 48),
Hodgkin's disease (55), and AIDS-associated B-cell
malignancies (1, 9, 16, 32). EBV-infected B-lymphoid cell
lines (B-LCLs) are infectable only at high particle/cell ratios of
Ad5-based vectors. This is likely due to their low level of CAR
expression, as these cells express elevated levels of We recently described a system consisting of fiber-expressing cell
lines and a fiber gene-deleted Ad vector (50, 51). Since the
fiber incorporated into such a vector during the last round of viral
growth need not bind the producing cells, this method will allow the
use of a much broader variety of fiber proteins in retargeting. While
the cells previously reported could complement a fiber gene-deleted
virus, the level of fiber expression varied from cell to cell and was
significantly below that seen in a normal infection (51). By
improving translational regulation of fiber expression in the packaging
cell lines, we have now increased the amount of fiber incorporated into
particles to near-wild-type levels. Pseudotyped Ad particles with
distinct cell tropisms were produced in packaging cell lines which
expressed two different fiber proteins (Fig.
1). In particular, Ad particles
containing a chimeric fiber protein with the receptor-binding domain of
Ad3 (46) infected EBV-infected B-LCLs much more efficiently
than standard Ad5 vectors.
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Adenovirus Vector Pseudotyping in Fiber-Expressing
Cell Lines: Improved Transduction of Epstein-Barr Virus-Transformed
B Cells
ABSTRACT
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
gal.
F) and packaging cells that express the viral fiber
protein. Expression of different fibers in packaging cells will allow
Ad retargeting without modifying the viral chromosome. Importantly,
fiber proteins which can no longer bind to the producer cells can also
be used. Using this approach, we generated for the first time
pseudotyped Ad5.gal.F particles containing either the wild-type
Ad5 fiber protein or a chimeric fiber with the receptor-binding knob
domain of the Ad3 fiber. Particles equipped with the chimeric fiber
bound to the Ad3 receptor rather than the coxsackievirus-adenovirus
receptor protein used by Ad5. EBV-transformed B lymphocytes were
infected efficiently by the Ad3-pseudotyped particles but poorly by
virus containing the Ad5 fiber protein. The strategy described here represents a broadly applicable method for targeting gene delivery to
specific cell types.
INTRODUCTION
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
v integrins (57). This is distinct from
fiber-receptor binding and appears to be conserved among many Ad
serotypes (35). Ads (including Ad5) of all subgroups except
subgroup B appear to use a fiber receptor termed coxsackievirus-Ad
receptor (CAR) (3, 41, 49). Subgroup B includes Ad3, which
has been shown to use a different fiber receptor (6, 46).
While CAR is widely expressed in vivo (49), low CAR levels
or its expression on an inaccessible part of the cell (such as the
basolateral surface of lung epithelial cells) can prevent efficient
transduction by Ad5 vectors (28, 38, 53). Replacement or
alteration of the fiber gene in the Ad chromosome has been shown to
alter viral tropism (12, 26, 45). In addition to the use of
natural variants, fiber proteins have been engineered to bind different receptors either by genetic modification or by antibody- or
ligand-mediated strategies (8, 13, 25, 26, 36, 40, 54, 58).
However, these modified vectors can be propagated only if their fiber
protein retains the ability to bind to the cells used for virus
production. This places a significant constraint on vector retargeting.
A method for producing virions that cannot bind their native receptors (e.g., CAR) would be more versatile.
v
integrins on their surfaces as a consequence of EBV infection
(17). More efficient transduction of these cells should be
possible through manipulation of the Ad fiber protein, facilitating the
development of effective therapies.
View larger version (22K):
[in a new window]
FIG. 1.
Strategy used for production of pseudotyped Ad vectors.
A fiber gene-deleted Ad vector such as Ad5. gal.F is grown in
packaging cell lines expressing different fiber proteins. The resulting
particles will have distinct cell tropisms.
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MATERIALS AND METHODS |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Cells and viruses.
THP-1, MRC-5, FaDu, and A-10 cells were
purchased from the American Type Culture Collection. 211B is a
293-derived cell line that expresses the wild-type Ad5 fiber protein
(51). E1-2a (14), an A549-derived cell line which
complements Ad E1 and E2a functions, was obtained from Michael Kadan,
Genetic Therapy, Inc. The JR, TO, and TL LCLs were established as
described previously (17) by EBV infection of lymphocytes
from three healthy donors. B-10 cells are a subclone of the JR LCL and
were produced by limiting dilution followed by PCR analysis to
determine loss of the EBV genome (S. Huang, unpublished data). THP-1,
all LCLs, and B-10 cells were maintained in RPMI 1640 medium
(Gibco)-10% fetal calf serum (FCS; Hyclone). 211B, MRC-5, and A-10
cells were grown in Dulbecco modified Eagle medium-10% FCS. E1-2a and
its derivatives were grown in Richter's modified medium (Bio
Whitaker)-10% FCS. Peripheral blood mononuclear cells (PBMCs) were
isolated from normal human blood (General Clinical Research Center,
Scripps Clinic) by sedimentation on Ficoll-Paque (Pharmacia) according to the manufacturer's instructions. Wild-type Ad2 and Ad3 were purchased from the American Type Culture Collection. Construction of
Ad5.gal.wt and Ad5.gal.F (50) has been previously
described. Av1LacZ4 (37) is a first- generation Ad5 vector
containing a Rous Sarcoma virus-driven -galactosidase reporter gene.
Av9LacZ4 (45) is identical to Av1LacZ4 except that the fiber
gene in the vector chromosome was replaced by a recombinant gene
encoding a chimeric fiber protein with the receptor-binding domain of
the Ad3 fiber (46).
DNA constructs.
The complete Ad5 tripartite leader (TPL)
contained in pDV67 and pDV69 was constructed by assembly of PCR
fragments. The third TPL exon (nucleotides [nt] 9644 to 9731 of the
Ad5 genome) was amplified by using the primers 5' CTC AAC AAT TGT
TGG ATC CGT ACT CC 3' and 5' GTG CTC AGC AGA TCT
TGC GAC TGT G 3'. The resulting product was cloned to the
BamHI and BglII sites of pE1Sp1a (Microbix
Biosystems) by using novel sites in the primers (in boldface) to create
pDV52. A fragment corresponding to the first TPL exon, the natural
first intron, and the second TPL exon (Ad5 nt 6049 to 7182) was
amplified by using primers 5' GGC GCG TTC GGA TCC ACT CTC
TTC C 3' and 5' CTA CAT GCT AGG CAG ATC TCG TTC GGA G 3' and
cloned into the BamHI site of pDV52, again using novel sites
in the primers (in boldface), to create pDV55. This plasmid contains a
1.2-kb BamHI/BglII fragment consisting of the
first TPL exon, the natural first intron, and the fused second and
third TPL exons. Finally, pDV60 was constructed by inserting this TPL
cassette into the BamHI site upstream of the Ad5 fiber gene
in pcDNA3/Fiber (51).
Construction of stable cell lines. E1-2a cells were electroporated as previously described (51) with pDV61, pDV67, or pDV69, and stable lines were selected with Zeocin (600 µg/ml; Invitrogen). Candidate clones were evaluated by immunofluorescence (51) using a polyclonal antibody generated against the Ad2 fiber (57). Those lines expressing the highest level of nuclear fiber were further characterized. Lines 601 and 633 were produced by transfection of pDV61 and pDV67, respectively, and therefore express the wild-type Ad5 fiber. Line 644 contains pDV69 and expresses the chimeric 5T3H fiber.
Virus growth and analysis. Ad stocks were prepared in the indicated cell lines and plaque titered on 633 cells essentially as described elsewhere (50). E1-2a cells (14) and their derivatives contain a dexamethasone-inducible construct for complementation of E1a. 601, 633, and 644 cells were therefore treated with 0.3 µM dexamethasone for 24 h prior to infection, and 0.5 µM dexamethasone was included in the overlay for plaque assays. Protein concentrations of viral preparations were determined by using the Bio-Rad protein assay with purified bovine serum albumin as a standard. Particle number was calculated by using the formula 1 µg of protein = 4 × 109 viral particles. Western blotting was performed as described elsewhere (51), using polyclonal rabbit antibodies raised against either the Ad2 (57) or Ad3 (45) fibers.
Infection and receptor binding assays.
Cells (2 × 105) in a total volume of 200 µl were incubated with the
indicated Ad preparation for 3 h at 37°C. Cells were then washed
twice with fresh medium and returned to 37°C. Two days later, cells
were fixed and stained with
5-bromo-4-chloro-3-indolyl--D-galactopyranoside (X-Gal)
and counted by light microscopy as described previously (50). For competition assays, cells were preincubated on ice for 1 h with either recombinant Ad3 fiber (10 µg/ml) purified from baculovirus or with a crude baculovirus lysate (100 µg/ml) containing the recombinant Ad2 fiber protein (57).
Expression of v integrins on cell surfaces was assayed
by fluorescence-activated cell sorting assay using monoclonal
antibodies (the gift of David Cheresh, Scripps Research Institute)
against either v3 (LM609) or
v5 (P1F6) as previously described
(17). For virus binding assays, CsCl-purified Ad2 or Ad3 was
labeled with 125I by using Iodogen tubes (Pierce). Free
iodine was removed by filtration with a PD-10 Sephadex column
(Pharmacia). Cells (106 cells in a volume of 200 µl
either with or without a 100-fold excess of unlabeled virus) were
rocked at 4°C for 2 h with 106 cpm of the labeled
virus, washed three times with phosphate-buffered saline, and counted.
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RESULTS |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Increased fiber expression in packaging lines. We previously reported the development of packaging cell lines that expressed Ad5 fiber at a level considerably below that seen in a normal infection (51). In an attempt to increase fiber production, we explored the use of additional viral regulatory elements. During a normal Ad infection, the translation of host cell proteins is inhibited. Viral proteins continue to be produced due to the action of the TPL, three small exons which are spliced onto the 5' end of late viral mRNAs. In addition to the leader's role in translational control, sequences in the first TPL intron have been reported to increase transcription from the viral major late promoter in Ad-infected cells (19, 29, 33, 34). Inclusion of a cassette containing a nearly complete TPL cDNA but lacking any TPL introns (43) allowed nuclear accumulation of fiber in packaging cell lines (51). We hypothesized that a more complete version of the TPL might improve fiber expression. A construct (pDV67) containing the entire first TPL intron, as well as complete copies of all three exons, was constructed and incorporated into expression constructs (Fig. 2A). Both versions of the fiber expression construct were electroporated into the E1a/E2a-complementing cell line E1-2a (14), and stable cell lines were isolated. Fiber expression was assayed by indirect immunofluorescence using a polyclonal antibody raised against the Ad2 fiber protein. As the Ad2 and Ad5 fibers are nearly identical, this antibody efficiently recognizes the Ad5 fiber protein. The improved TPL resulted both in a generally higher level of fiber protein expression and in a much smaller number of low-expressing cells (Fig. 2). One fiber-expressing clone carrying each construct (lines 601 and 633 contain the original and improved constructs, respectively) was selected for further evaluation.
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gal.F
(50) was used to assess fiber complementation by the
various cell lines. We prepared stocks of this virus in cells
expressing the wild-type Ad5 fiber under the control of either the
partial Ad TPL contained in pDV61 (line 601) or the complete TPL plus intron as in pDV67 (line 633). As a control, fiber levels in these preparations were compared to the level in the first-generation vector Ad5.gal.wt, which is identical to Ad5.gal.F
except for the fiber deletion. Consistent with the increased
level of fiber synthesis in line 633, Ad5.gal.F grown in these
cells contained a much larger (nearly normal) amount of fiber protein
than those produced in line 601 (Fig. 2D).
Particle yields of Ad5.gal.F were comparable to those of
Ad5.gal.wt (Table 1). However, the
infectious titers of the viral preparations were quite different. Virus
produced in cells (line 601) expressing fiber from the incomplete TPL
construct was much less infectious than the control, as seen by the
increased particle/PFU and particle/-galactosidase transducing unit
(TU) ratios (Table 1). As well as increasing the amount of fiber on the
particles, growth of Ad5.gal.F in cells carrying the improved
expression construct (line 633) partially rescued the defect in plaque
formation. Interestingly, while the number of particles per PFU was
still approximately 70-fold higher than for a first-generation Ad
vector, LacZ transduction of 293 cells was now similar to (within
fourfold of) normal (Table 1; compare preparations 1 and 2 to
preparations 7 and 8). This may indicate that deletion of the fiber
gene affects late events in the virus life cycle, rather than early
events such as gene delivery to the nucleus (see Discussion). Together, these results demonstrated that the reduced infectivity of
Ad5.gal.F grown in cells (such as 211B or 601) expressing fiber
under the control of the partial TPL was at least partly due to a
deficit of fiber, as increased fiber content on particles due to the
improved expression construct correlated with improved infectivity.
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Pseudotyping of an Ad vector. In addition to producing vectors with the wild-type Ad5 tropism, the packaging cell technology should allow retargeting by pseudotyping a vector with modified fiber proteins or with fibers from different Ad serotypes. To test this hypothesis, we used the chimeric 5T3H fiber described above (46). Virus particles containing this fiber would be expected to bind and infect cells via the Ad3 receptor rather than the CAR protein used by Ad5.
Growth of Ad5.gal.F in cells (line 644) expressing the chimeric
fiber produced yields of viral particles similar to those seen with the
other packaging lines (Table 1), and immunoblot analysis (Fig.
3A) demonstrated that they contained high
levels of the chimeric fiber protein. Purified particles of Av9LacZ4, which is an E1-deleted virus containing the chimeric 5T3H gene in its
chromosome (45), and of Ad5.gal.wt were analyzed as positive controls. While both the wild-type Ad5 and chimeric 5T3H fibers could be detected by a polyclonal antibody raised against the
Ad2 fiber, an anti-Ad3 fiber antibody detected only the chimeric fiber
by virtue of its Ad3 knob domain. As a control for protein loading, the
membrane was reprobed with a polyclonal antibody directed against the
Ad2 penton base protein (which cross-reacts with the Ad5 penton base
protein).
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gal.F viral particles containing the chimeric
fiber protein were slightly more infectious than those equipped with
the Ad5 fiber (Table 1). This might reflect either a somewhat higher
level of the Ad3 receptor on 633 cells or more efficient complementation of the fiber deletion. Since comparing plaque titers of
virions which use different attachment receptors may be misleading, we
have given all multiplicities of infection as the number of physical
particles per cell. Receptor usage by the pseudotyped Ad5.gal.F
was assessed in competition experiments. 211B cells (which express both
CAR and the Ad3 receptor) were infected in the presence or absence of
excess recombinant Ad2 or Ad3 fiber proteins (Fig. 3B). Addition of
recombinant Ad2 fiber completely blocked infection by virus containing
the Ad5 fiber protein but not by Ad3-pseudotyped virus. Conversely, an
excess of recombinant Ad3 fiber which abolished infection by the
Ad3-pseudotyped particles had no effect on those containing the Ad5
fiber. Consistent with the role of v integrins in
infection and internalization of both Ad3 and Ad5 (35),
infection by either preparation of Ad5.gal.F was blocked by
addition of excess recombinant Ad2 penton base protein.
Altered in vitro tropism and infection of B-LCLs.
Experiments
with genetically modified viruses showed that a number of different
cell types are more readily infected through interaction with the Ad3
receptor than by the CAR-dependent pathway used by Ad5 (45).
To further evaluate our pseudotyping system, we assayed the ability of
Ad5.gal.F carrying either the Ad5 or chimeric 5T3H fibers to
infect several of these: FaDu (a head and neck tumor line), THP-1
monocytic cells, and MRC-5 fibroblasts. Consistent with the previous
studies (45), use of the chimeric Ad5-Ad3 fiber protein
increased infection of all of these lines at equal particle/cell ratios
(Fig. 4). In contrast, the rat smooth muscle cell line A-10 was infected somewhat more readily by Ad5- than
by Ad3-pseudotyped particles (Fig. 4), also in agreement with previous
results (S. C. Stevenson, unpublished data).
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(cpm bound with
competitor). Values reported are means of duplicate samples and are
representative of several experiments. (C) v integrins on all four cell lines by
fluorescence-activated cell sorting using antibodies specific for
v5 or v3
integrins. All of the LCLs which supported infection expressed both
v5 and v3
integrins, while B-10 cells expressed neither (Fig. 5C). This finding
emphasizes that expression of both a fiber receptor and of
v integrins are required for efficient Ad infection.
Selective gene delivery to EBV-infected cells.
The results
above suggested that the minority of EBV-infected B cells present in
donor marrow or peripheral blood would be preferentially infected by
vectors using the Ad3 receptor. To test this hypothesis, we performed a
mixing experiment with uninfected PBMCs and EBV-infected cells. JR LCLs
were mixed at various ratios with PBMCs isolated from a healthy human
donor, and the mixture was then infected with Ad5.gal.F particles
containing the 5T3H fiber protein. No infection of normal PBMCs alone
was detected. Moreover, the percent of total cells infected increased
with the fraction of JR cells added (Fig.
6). This finding indicates that EBV-infected cells can be selectively infected in vitro by relatively short (3-h) exposure to a retargeted Ad vector.
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DISCUSSION |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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We previously reported a fiber gene-deleted Ad5 vector and its growth in fiber-expressing packaging cell lines. The data presented here demonstrate that such a vector can be retargeted simply by production in packaging lines expressing different fiber proteins. A wide variety of fibers, including those which cannot bind to the host cells used for virus production, should be useful in such cell lines. This is especially important since developing viruses targeted to infect a single cell type would involve eliminating their binding to the natural fiber receptor (CAR). Our system will also simplify development of Ad-mediated therapies aimed at different applications. Rather than generating a new viral chromosome each time a transgene is to be delivered to a new target, a single vector could be targeted to different cell populations by pseudotyping with the appropriate fiber proteins. The cell lines described here were derived from the very efficient E1a/E2a-complementing line (14) and should allow the future development of Ad vectors with deletions of E1a, E3, L5 (fiber), and E2a.
Our original generation of packaging cell lines (51) expressed the Ad5 fiber at a relatively low level. They produced Ad particles containing a substoichiometric amount of fiber protein, with reduced infectivity relative to first-generation vector particles (50). This appears to be a property of the fiber expression construct used to generate these lines. While inclusion of an incomplete TPL fragment greatly increased fiber expression over that seen from a construct lacking TPL sequences, there was a large amount of cell-to-cell variation in fiber expression (51). Attempts to isolate cells that expressed fiber more uniformly by recloning line 211B were unsuccessful (data not shown).
In addition to the TPL's role in translational regulation, its natural first intron has been reported to act as a transcriptional enhancer for the Ad major late promoter (19, 29, 33, 34). We found that inclusion of this intron could also greatly increase fiber production from our cytomegalovirus-driven construct. Cell lines (such as line 633) carrying this construct not only expressed more fiber protein but exhibited much less cell-to-cell variability in expression level. Since synthesis of the major Ad structural proteins is coordinately regulated, this type of construct may be useful in developing systems for complementation of other viral proteins such as penton and hexon.
Ad5.gal.F growth in the improved packaging lines resulted in a
near-normal fiber content on the particles and improved infectivity severalfold relative to particles grown in cells carrying the original
expression construct. Interestingly, while the ability of
Ad5.gal.F particles produced in the new cell lines to transduce LacZ is very close to the wild-type level (Table 1), the particle/PFU ratio remains significantly (approximately 70-fold) higher. This suggests that there is an additional defect associated with the fiber
gene deletion, beyond a simple deficit of fiber protein, which is not
efficiently complemented by the packaging system that we are using. The
fact that plaque formation but not gene delivery is reduced suggests
that this defect might affect a stage in the viral life cycle
occurring after delivery of the viral chromosome to the nucleus.
In this light, it is interesting that Yeh and coworkers (59)
reported similar findings in work with an E4-lacking Ad5 vector.
There were several previous reports (4, 7, 10) of fiber
mutants being associated with defects in viral assembly or maturation,
but we did not detect obvious assembly problems with Ad5.gal.F
even in the complete absence of fiber. Indeed, a cryoelectron microscopic analysis of our fiberless particles showed their structure to be essentially normal (50). However, another group
recently reported production of a fiber gene-deleted Ad vector and did find differences in several aspects of the viral biology
(27). In addition to the expected reduced infectivity, they
reported slight differences in maturation of some capsid proteins as
determined by pulse-labeling with [35S]methionine,
increased cytoplasmic versus nuclear localization of the particles, and
altered particle density on CsCl gradients (27). It is
possible that while the defects detected by LeGrand et al.
(27) are functionally significant, they were not severe enough to have been detected by the methods that we used. We are now
examining the phenotype of our vector particles in more detail in order
to understand the differences in our results.
Use of the chimeric 5T3H fiber protein generated Ad5 vector particles
which infected cells via the Ad3 receptor rather than the CAR protein
used by Ad5 (Fig. 3 and 4). This is in agreement with previous data
showing that an Ad5 vector with this chimeric fiber gene substituted
into its chromosome displayed Ad3 tropism (45). The
retargeted virus remained dependent on the integrin-penton base
interaction, since infection by particles containing either fiber was
blocked by competition with excess recombinant Ad2 penton base. We also
found that the LCL-derived B-10 cell line, which lacks detectable
v integrins, was refractory to infection even though the
cells could bind labeled Ad3 particles.
EBV-infected B lymphocytes are logical therapeutic targets for a number
of diseases such as AIDS-associated central nervous system lymphoma or
transplant-associated lymphoproliferation. Although uninfected B
cells are not efficiently infected by Ad due to lack of integrin
expression, the upregulation of v integrins induced by
EBV infection allows virus internalization and gene delivery
(17) if very high particle/cell ratios of an Ad5 vector are
used. This inefficient infection is likely due to the low level of CAR
expression which we and others (47) have found on these
cells. As would be predicted from the binding data shown here, we found
that LCLs were much more readily infected by vector particles that
contained the Ad3 fiber knob. Our results are in agreement with a
previous report (5) showing that LCLs could be transduced by
an Ad-polylysine complex (in which the Ad moiety mediated endosome
disruption) provided that the complex was able to bind to the cells.
Interestingly, recent studies have detected replication of subgroup B
(the subgroup which includes Ad3) Ads, notably Ad35, in patients who
are immunosuppressed due to transplantation or to AIDS (31).
These serotypes are rarely found in healthy individuals. EBV infection
is often reactivated in such patients (48), and it is
possible that the EBV-induced upregulation of integrins allows
productive infection of lymphocytes by these Ad serotypes. In
preliminary studies, we have found that wild-type Ad3 is indeed capable
of infecting and replicating in B-LCLs (D. J. Von Seggern
and S. K. Fleck, unpublished data).
The differential infectivities of normal and EBV-infected B cells may be useful in such gene therapy strategies as purging donor bone marrow of EBV-infected cells before transplantation or the in vivo treatment of EBV-induced lymphomas. Previous work in our group has shown that introduction of a ribozyme targeted against the EBNA-1 transcript can reduce the EBV genome to undetectable levels in LCLs (17). Gene therapy strategies for these diseases based on specific transcription of suicide genes driven by EBV-responsive promoters have also been proposed (11, 21, 22, 42). Combining selective infection of EBV-positive cells by retargeted Ad vectors with such EBV-targeted therapeutic strategies might provide effective and specific treatments.
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ACKNOWLEDGMENTS |
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This work was supported by NIH grants EY11431 and HL54352 to G. R. Nemerow and by GTI/Novartis grant SFP1089 to D. J. Von Seggern and G. R. Nemerow.
We thank Joan Gausepohl for assistance with the manuscript, and we thank Phyllis Frosst and Colleen McKiernan for their comments. We also thank David Cheresh for his gift of monoclonal antibodies. Normal human donor blood was obtained from the GCRC under protocol 95-13.
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FOOTNOTES |
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* Corresponding author. Mailing address: Department of Immunology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-8072. Fax: (858) 784-8472. E-mail: gnemerow{at}scripps.edu.
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REFERENCES |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
| 1. |
Bashir, R.,
J. Luka,
K. Cheloha,
M. Chamberlain, and F. Hochberg.
1993.
Expression of Epstein-Barr virus proteins in primary CNS lymphoma in AIDS patients.
Neurology
43:2358-2361 |
| 2. | Bergelson, J. M. 1999. Receptors mediating adenovirus attachment and internalization. Biochem. Pharmacol. 57:975-979[CrossRef][Medline]. |
| 3. |
Bergelson, J. M.,
J. A. Cunningham,
G. Droguett,
E. A. Kurt-Jones,
A. Krithivas,
J. S. Hong,
M. S. Horwitz,
R. L. Crowell, and R. W. Finberg.
1997.
Isolation of a common receptor for coxsackie B viruses and adenoviruses 2 and 5.
Science
275:1320-1323 |
| 4. |
Chee-Sheung, C. C., and H. S. Ginsberg.
1982.
Characterization of a temperature-sensitive fiber mutant of type 5 adenovirus and effect of the mutation on virion assembly.
J. Virol.
42:932-950 |
| 5. | Curiel, T. J., D. R. Cook, C. Bogedain, W. Jilg, G. S. Harrison, M. Cotten, D. T. Curiel, and E. Wagner. 1999. Efficient foreign gene expression in Epstein-Barr virus-transformed human B-cells. Virology 198:577-585. |
| 6. |
Defer, C.,
M.-T. Belin,
M.-L. Caillet-Boudin, and P. Boulanger.
1990.
Human adenovirus-host cell interactions: comparative study with members of subgroups B and C.
J. Virol.
64:3661-3673 |
| 7. |
D'Halluin, J.-C.,
M. Milleville,
G. R. Martin, and P. Boulanger.
1980.
Morphogenesis of human adenovirus type 2 studied with fiber- and fiber and penton base-defective temperature-sensitive mutants.
J. Virol.
33:88-99 |
| 8. |
Dmitriev, I.,
V. Krasnykh,
C. R. Miller,
M. Wang,
E. Kashentseva,
G. Mikheeva,
N. Belousova, and D. T. Curiel.
1998.
An adenovirus vector with genetically modified fibers demonstrates expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent cell entry mechanism.
J. Virol.
72:9706-9713 |
| 9. | Ernberg, I., and E. Altiok. 1989. The role of Epstein-Barr virus in lymphomas of HIV carriers. APMIS 97:58-61. |
| 10. |
Falgout, B., and G. Ketner.
1988.
Characterization of adenovirus particles made by deletion mutants lacking the fiber gene.
J. Virol.
62:622-625 |
| 11. | Franken, M., A. Estabrooks, L. Cavacini, B. Sherburne, F. Wang, and D. T. Scadden. 1996. Epstein-Barr virus-driven gene therapy for EBV-related lymphomas. Nat. Med. 2:1379-1382[CrossRef][Medline]. |
| 12. | Gall, J., A. Kass-Eisler, L. Leinwand, and E. Falck-Pedersen. 1996. Adenovirus type 5 and 7 capsid chimera: fiber replacement alters receptor tropism without affecting primary immune neutralization epitopes. J. Virol. 70:2116-2123[Abstract]. |
| 13. |
Goldman, C. K.,
B. E. Rogers,
J. T. Douglas,
B. A. Sosnowski,
W. Ying,
G. P. Siegal,
A. Baird,
J. A. Campain, and D. T. Curiel.
1997.
Targeted gene delivery to Kaposi's sarcoma cells via the fibroblast growth factor receptor.
Cancer Res.
57:1447-1451 |
| 14. | Gorziglia, M. I., M. J. Kadan, S. Yei, J. Lim, G. M. Lee, R. Luthra, and B. C. Trapnell. 1996. Elimination of both E1 and E2a from adenovirus vectors further improves prospects for in vivo human gene therapy. J. Virol. 70:4173-4178[Abstract]. |
| 15. | Haddada, H., L. Cordier, and M. Perricaudet. 1995. Gene therapy using adenovirus vectors. Curr. Top. Microbiol. Immunol. 199:297-306. |
| 16. |
Hamilton-Dutoit, S. J.,
M. Raphael,
J. Audouin,
J. Diebold,
I. Lisse,
C. Pedersen,
E. Oksenhendler,
L. Marelle, and G. Pallesen.
1993.
In situ demonstration of Epstein-Barr virus small RNAs (EBER 1) in acquired immunodeficiency syndrome-related lymphomas: correlation with tumor morphology and primary site.
Blood
82:619-625 |
| 17. |
Huang, S.,
D. G. Stupack,
P. Mathias,
Y. Wang, and G. Nemerow.
1997.
Growth arrest of Epstein-Barr virus immortalized B lymphocytes by adenovirus-delivered ribozymes.
Proc. Natl. Acad. Sci. USA
94:8156-8161 |
| 18. | Huard, J., H. Lochmüller, G. Acsadi, A. Jani, B. Massie, and G. Karpati. 1995. The route of administration is a major determinant of the transduction efficiency of rat tissues by adenoviral recombinants. Gene Ther. 2:107-115[Medline]. |
| 19. |
Jansen-Durr, P.,
H. Boeuf, and C. Kédinger.
1988.
Replication-induced stimulation of the major late promoter of adenovirus is correlated to the binding of a factor to sequences in the first intron.
Nucleic Acids Res.
16:3771-3786 |
| 20. |
Jooss, K.,
Y. Yang,
K. J. Fisher, and J. M. Wilson.
1998.
Transduction of dendritic cells by DNA viral vectors directs the immune response to transgene products in muscle fibers.
J. Virol.
72:4212-4223 |
| 21. | Judde, J.-G., G. Spangler, I. Magrath, and K. Bhatia. 1996. Use of Epstein-Barr virus nuclear antigen-1 in targeted therapy of EBV-associated neoplasia. Hum. Gene Ther. 7:647-653[Medline]. |
| 22. | Kenney, S., J.-Q. Ge, E. M. Westphal, and J. Olsen. 1998. Gene therapy strategies for treating Epstein-Barr virus-associated lymphomas: comparison of two different Epstein-Barr virus-based vectors. Hum. Gene Ther. 9:1131-1141[Medline]. |
| 23. | Kovesdi, I., D. E. Brough, J. T. Bruder, and T. J. Wickham. 1997. Adenoviral vectors for gene transfer. Curr. Opin. Biotechnol. 8:583-589[CrossRef][Medline]. |
| 24. | Kozarsky, K. F., and J. M. Wilson. 1993. Gene therapy: adenovirus vectors. Curr. Opin. Genet. Dev. 3:499-503[CrossRef][Medline]. |
| 25. |
Krasnykh, V.,
I. Dmitriev,
G. Mikheeva,
C. R. Miller,
N. Belousova, and D. T. Curiel.
1998.
Characterization of an adenovirus vector containing a heterologous peptide epitope in the HI loop of the fiber knob.
J. Virol.
72:1844-1852 |
| 26. |
Krasnykh, V. N.,
G. V. Mikheeva,
J. T. Douglas, and D. T. Curiel.
1996.
Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism.
J. Virol.
70:6839-6846 |
| 27. |
Legrand, V.,
D. Spehner,
Y. Schlesinger,
N. Settelen,
A. Pavirani, and M. Mehtali.
1999.
Fiberless recombinant adenoviruses: virus maturation and infectivity in the absence of fiber.
J. Virol.
73:907-919 |
| 28. |
Leon, R. P.,
T. Hedlund,
S. J. Meech,
S. Li,
J. Schaack,
S. P. Hunger,
R. C. Duke, and J. DeGregori.
1998.
Adenoviral-mediated gene transfer in lymphocytes.
Proc. Natl. Acad. Sci. USA
95:13159-13164 |
| 29. |
Leong, K.,
W. Lee, and A. J. Berk.
1990.
High-level transcription from the adenovirus major late promoter requires downstream binding sites for late-phase-specific factors.
J. Virol.
64:51-60 |
| 30. | Lucas, K. G., K. E. Pollok, and D. J. Emanuel. 1997. Post-transplant EBV induced lymphoproliferative disorders. Leuk. Lymphoma 25:1-8[Medline]. |
| 31. | Lukashok, S. A., and M. S. Horwitz. 1998. New perspectives in adenovirus. Curr. Clin. Top. Infect. Dis. 18:286-304[Medline]. |
| 32. | MacMahon, E. M. E., J. D. Glass, S. D. Hayward, R. B. Mann, P. S. Becker, P. Charache, J. C. McArthur, and R. F. Ambinder. 1991. Epstein-Barr virus in AIDS-related primary central nervous system lymphoma. Lancet 338:969-973[CrossRef][Medline]. |
| 33. |
Mansour, S. L.,
T. Grodzicker, and R. Tjian.
1986.
Downstream sequences affect transcription initiation from the adenovirus major late promoter.
Mol. Cell. Biol.
6:2684-2694 |
| 34. | Mason, B. B., A. R. Davis, B. M. Bhat, M. Chengalvala, M. D. Lubeck, G. Zandle, B. Kostek, S. Cholodofsky, S. Dheer, K. Molnar-Kimber, S. Mizutani, and P. P. Hung. 1990. Adenovirus vaccine vectors expressing hepatitis B surface antigen: importance of regulatory elements in the adenovirus major late intron. Virology 177:452-461[CrossRef][Medline]. |
| 35. |
Mathias, P.,
T. J. Wickham,
M. Moore, and G. Nemerow.
1994.
Multiple adenovirus serotypes use v integrins for infection.
J. Virol.
68:6811-6814 |
| 36. | Michael, S. I., J. S. Hong, D. T. Curiel, and J. A. Engler. 1995. Addition of a short peptide ligand to the adenovirus fiber protein. Gene Ther. 2:660-668[Medline]. |
| 37. | Mittereder, N., K. L. March, and B. C. Trapnell. 1996. Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J. Virol. 70:7498-7509[Abstract]. |
| 38. | Nalbantoglu, J., G. Pari, G. Karpati, and P. C. Holland. 1999. Expression of the primary coxsackie and adenovirus receptor is downregulated during skeletal muscle maturation and limits the efficacy of adenovirus-mediated gene delivery to muscle cells. Hum. Gene Ther. 10:1009-1019[CrossRef][Medline]. |
| 39. |
Philipson, L.,
K. Lonberg-Holm, and U. Pettersson.
1968.
Virus-receptor interaction in an adenovirus system.
J. Virol.
2:1064-1075 |
| 40. |
Rancourt, C.,
B. E. Rogers,
B. A. Sosnowski,
M. Wang,
A. Piché,
G. F. Pierce,
R. D. Alvarez,
G. P. Siegal,
J. T. Douglas, and D. T. Curiel.
1998.
Basic fibroblast growth factor enhancement of adenovirus-mediated delivery of the herpes simplex virus thymidine kinase gene results in augmented therapeutic benefit in a murine model of ovarian cancer.
Clin. Cancer Res.
4:2455-2461 |
| 41. |
Roelvink, P. W.,
A. Lizonova,
J. G. M. Lee,
Y. Li,
J. M. Bergelson,
R. W. Finberg,
D. E. Brough,
I. Kovesdi, and T. J. Wickham.
1998.
The coxsackievirus-adenovirus receptor protein can function as a cellular attachment protein for adenovirus serotypes from subgroups A, C, D, E, and F.
J. Virol.
72:7909-7915 |
| 42. | Rogers, R. P., J.-Q. Ge, E. Holley-Guthrie, D. K. Hoganson, K. E. Comstock, J. C. Olsen, and S. Kenney. 1996. Killing Epstein-Barr virus-positive B lymphocytes by gene therapy: comparing the efficacy of cytosine deaminase and herpes simplex virus thymidine kinase. Hum. Gene Ther. 7:2235-2245[Medline]. |
| 43. | Sheay, W., S. Nelson, I. Martinez, T.-H. T. Chu, S. Bhatia, and R. Dornburg. 1993. Downstream insertion of the adenovirus tripartite leader sequence enhances expression in universal eukaryotic vectors. Bio Techniques 15:856-862. |
| 44. | Smith, T. A. G., M. G. Mehaffey, D. B. Kayda, J. M. Saunders, S. Yei, B. C. Trapnell, A. McClelland, and M. Kaleko. 1993. Adenovirus mediated expression of therapeutic plasma levels of human factor IX in mice. Nat. Genet. 5:397-402[CrossRef][Medline]. |
| 45. | Stevenson, S. C., M. Rollence, J. Marshall-Neff, and A. McClelland. 1997. Selective targeting of human cells by a chimeric adenovirus vector containing a modified fiber protein. J. Virol. 71:4782-4790[Abstract]. |
| 46. | Stevenson, S. C., M. Rollence, B. White, L. Weaver, and A. McClelland. 1995. Human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain. J. Virol. 69:2850-2857[Abstract]. |
| 47. |
Teoh, G.,
L. Chen,
M. Urashima,
Y.-T. Tai,
L. A. Celi,
D. Chen,
D. Chauhan,
A. Ogata,
R. W. Finberg,
I. J. Webb,
D. W. Kufe, and K. C. Anderson.
1998.
Adenovirus vector-based purging of multiple myeloma cells.
Blood
92:4591-4601 |
| 48. | Thomas, J. A., M. J. Allday, and D. H. Crawford. 1991. Epstein-Barr virus-associated lymphoproliferative disorders in immunocompromised individuals. Adv. Cancer Res. 57:329-380[Medline]. |
| 49. |
Tomko, R. P.,
R. Xu, and L. Philipson.
1997.
HCAR and MAR: the human and mouse cellular receptors for subgroup C adenoviruses and group B coxsackieviruses.
Proc. Natl. Acad. Sci. USA
94:3352-3356 |
| 50. |
Von Seggern, D. J.,
C. Y. Chiu,
S. K. Fleck,
P. L. Stewart, and G. R. Nemerow.
1999.
A helper-independent adenovirus vector with E1, E3, and fiber deleted: structure and infectivity of fiberless particles.
J. Virol.
73:1601-1608 |
| 51. | Von Seggern, D. J., J. Kehler, R. Endo, and G. R. Nemerow. 1998. Complementation of a fiber mutant adenovirus by packaging cell lines stably expressing the Ad5 fiber protein. J. Gen. Virol. 79:1461-1468[Abstract]. |
| 52. | Von Seggern, D. J., and G. R. Nemerow. 1999. Adenoviral vectors for protein expression, p. 111-156. In J. Fernandez, and J. Hoeffler (ed.), Gene expression systems: using nature for the art of expression. Academic Press, San Diego, Calif |
| 53. |
Walters, R. W.,
T. Grunst,
J. M. Bergelson,
R. W. Finberg,
M. J. Welsh, and J. Zabner.
1999.
Basolateral localization of fiber receptors limits adenovirus infection from the apical surface of airway epithelia.
J. Biol. Chem.
274:10219-10226 |
| 54. | Watkins, S. J., V. V. Mesyanzhinov, L. P. Kurochkina, and R. E. Hawkins. 1997. The 'adenobody` approach to viral targeting: specific and enhanced adenoviral gene delivery. Gene Ther. 4:1004-1012[CrossRef][Medline]. |
| 55. | Weiss, L. M., L. A. Movahed, R. A. Warnke, and J. Sklar. 1989. Detection of Epstein-Barr viral genomes in Reed-Sternberg cells of Hodgkin's disease. N. Engl. J. Med. 320:502-506[Abstract]. |
| 56. |
Westphal, E.-M.,
A. Mauser,
J. Swenson,
M. G. Davis,
C. L. Talarico, and S. C. Kenney.
1999.
Induction of lytic Epstein-Barr virus (EBV) infection in EBV-associated malignancies using adenovirus vectors in vitro and in vivo.
Cancer Res.
59:1485-1491 |
| 57. |
Wickham, T. J.,
P. Mathias,
D. A. Cheresh, and G. R. Nemerow.
1993.
Integrins v3 and v5 promote adenovirus internalization but not virus attachment.
Cell
73:309-319[CrossRef][Medline].
|
| 58. | Wickham, T. J., E. Tzeng, L. L. Shears II, P. W. Roelvink, Y. Li, G. M. Lee, D. E. Brough, A. Lizonova, and I. Kovesdi. 1997. Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins. J. Virol. 71:8221-8229[Abstract]. |
| 59. | Yeh, P., J.-F. Dedieu, C. Orsini, E. Vigne, P. Denefle, and M. Perricaudet. 1996. Efficient dual transcomplementation of adenovirus E1 and E4 regions from a 293-derived cell line expressing a minimal E4 functional unit. J. Virol. 70:559-565[Abstract]. |
| 60. | Zhong, L., A. Granelli-Piperno, Y. Choi, and R. M. Steinman. 1999. Recombinant adenovirus is an efficient and non-perturbing genetic vector for human dendritic cells. Eur. J. Immunol. 29:964-972[CrossRef][Medline]. |
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