вход по аккаунту


In vivo suppression of early experimental osteoarthritis by interleukin-1 receptor antagonist using gene therapy.

код для вставкиСкачать
Vol. 40, No. 6, June 1997, pp 1012-1019
0 1993, American College of Rheumatology
Objective. This study explored the therapeutic
effect of interleukin-1 receptor antagonist (IL-lRa),
administered by gene transfer, on the progression of
osteoarthritic (OA) lesions in an experimental dog
Methods. Seventeen mature mongrel dogs were
divided into 3 groups. Group 1 (n = 7) had an anterior
cruciate ligament (ACL) section of the right knee
through a stab wound incision. Groups 2 and 3 (n = 5
per group), had an ACL section of the right knee and
partial synovectomy of the left knee. Each dog’s synovium was subjected to enzymatic digestion, and the
synovial fibroblasts were propagated in monolayer culture. Synovial cells from each dog were transduced in
vitro using the retrovirus MFG with either the Escherichia coli P-galactosidase (lac Z) gene (group 2) or the
human IL-1Ra gene (group 3). Two days after surgery,
the dogs received intraarticular injections as follows:
group 1phosphate buffered saline (PBS) (2 ml); group
2 autologous cells (60 x lo6 cells/2 ml of PBS) transduced with the lac Z gene; group 3 autologous cells
transduced with the IL-1Ra gene. Synovial fluid was
aspirated at 2 weeks and 4 weeks. All dogs were euthanized
at 4 weeks postsurgery. The right knees were dissected,
and lesions were scored for macroscopic and microscopic changes. Synovial explants were dissected and
representative specimens were used for histology or were
cultured for 48 hours. The levels of I GlR a in synovial fluid
and synovial explant conditioned medium were measured
by specific enzyme-linked immunosorbent assay.
Results. The level of IL-1Ra in synovial fluid of
group 3 was 202.8 f 131.5 ng/ml (mean f SEM) at 2
weeks and 2.8 f 2.2 ng/ml a t 4 weeks after surgery.
Membrane explants isolated from dogs that received
synovial cells transduced with the IL-1Ra gene (group 3)
actively produced IL-1Ra (4.0 f 2.0 ng/gm of tissue wet
weight). The severity of OA cartilage lesions was similar
in groups 1 and 2. In contrast, group 3 dogs had a
marked reduction in macroscopic lesion severity on the
tibia1 plateaus (P c 0.01 for grade; P c 0.04 for size)
and femoral condyles. Moreover, the histologic lesion
severity was decreased on both plateaus (Pc 0.06) and
Conclusion. This study showed that a local
increase in IL-1Ra production in OA knee joints
by intraarticular injection of transduced synovial cells
can reduce the progression of experimentally induced
Supported by grants from the Medical Research Council of
Canada and NIH grant no. 44935.
Jean-Pierre Pelletier, MD, Julio C. Fernandes, MD, MSc,
Johannc Martel-Pelletier, PhD: University of Montreal, L.-C. Simard
Research Center, and Notre-Dame Hospital, Montreal, Quebec, Canada;
John P. Caron, DVM, MVSc: L.-C. Simard Research Center, NotreDame Hospital, Montreal, Quebec, Canada, and Michigan State
University, East Lansing; Christopher Evans, PhD, Paul D. Robbins,
PhD, Helga I. Georgescu. BSc: University of Pittsburgh, Pittsburgh,
Pennsylvania; Dragan Jovanovic, MD, PhD: L.-C. Simard Research
Center and Notre-Dame Hospital, Montreal, Quebec, Canada.
Address reprint requests to Jean-Pierre Pelletier, MD, Director, Osteoarthritis Research Unit, L.-C. Simard Research Centre,
Notre-Dame Hospital, 1560 Sherbrookc East, Montreal, Quebec, H2L
4M1 Canada.
Submitted for publication November 13, 1996; accepted in
revised form January 22, 1997.
Osteoarthritis (OA) is a degenerative joint disease with morphologic changes such as cartilage erosion,
osteophyte formation, subchondral bone remodeling,
and synovial inflammation. Although this disease might
arise from multiple origins, evidence emphasizes that
both mechanical and biochemical forces are leading
causes of its appearance and/or progression (1).
Recent advances in OA research point to important biochemical pathways as key factors in cartilage
matrix breakdown. At the clinical stage of the disease,
there is synovial inflammation that is believed to be
secondary to the release of these products. A lively
interaction between the synovial membrane and the
cartilage prevail at this time, which further enhances
cartilage degradation (2). Morphologic changes of the
inflamed synovium include synovial lining cell hypertrophy and hyperplasia, as well as cellular infiltration,
consisting of a mixed population of mononuclear cells,
into the sublining tissue. In OA, however, it appears that
the lining cells, in particular, type B cells with a
fibroblast-like structure, are the site of important synthesis of proteases, such as matrix metalloproteases
(MMP), as well as proinflammatory cytokines (2,3).
Among the proinflammatory cytokines, interleukin-1
(IL-1) is the most abundantly synthesized by the OA
synovium (2,4) and appears to be primarily responsible
for inducing protease synthesis and cartilage catabolism.
Its biologic action on OA tissue is enhanced by at least
2 principal factors: an increased number of IL-1 receptors (IL-1R) by OA synovial fibroblasts and chondrocytes (5,6), and a relative deficit in the level of a natural
IL-1R antagonist (IL-1Ra) (4,7). Interestingly, we recently reported that, in these pathologic tissues, the
increased production of nitric oxide (NO) induced a
decreased synthesis of IL-1Ra (S), although the exact
reason for this phenomenon remains to be determined.
There are several possible strategies for controlling the biologic activity of IL-1 in the articular joint
tissues. Inhibition of its action at the cell receptor level
is one of them, and the IL-1Ra molecule appeared a
likely candidate. In vitro, this molecule has proven its
capacity to inhibit not only MMP synthesis, but also
cartilage degradation (9-1 1). Moreover, recent ex vivo
and in vivo experiments have shown cartilage protection
with IL-1Ra. Baragi et a1 (12), using an OA cartilage
explant system in which transplantation of chondrocytes
transduced with the IL-1Ra gene, showed that this
molecule protected articular cartilage from IL-1induced matrix degradation. Furthermore, we demonstrated that intraarticular injections of recombinant human IL-1Ra (rHuIL-1Ra) into the OA knee of an
experimental dog model decreased the progression of
OA cartilage lesions (13).
Although it was shown that this molecule could
be delivered by direct injection, the short intraarticular
half-life (a few hours) necessitated repeated injections.
The rapid clearance of IL-1Ra was also a problem in a
recent rheumatoid arthritis (RA) clinical trial in which
patients had to give themselves daily subcutaneous injections of IL-1Ra (14). Gene therapy, however, might
prove to be a suitable alternative. Interestingly, it has
been shown that increased local intraarticular synthesis
of antiarthritic proteins such as IL-1Ra can be achieved
by synovial delivery of the cognate genes (15-17). By
engineering the diseased joint in this way to become the
site of synthesis of its own drug, protein delivery problems are overcome, side effects are minimized, exposure
of nontarget organs is reduced, and the protein expression is prolonged. We therefore pursued our work on
IL-1Ra by exploring whether, when autografted by intraarticular injection, the rHuIL-1Ra gene-transduced
dog OA synovial fibroblasts could influence the course
of early OA.
Experimental groups. Seventeen adult crossbred
dogs weighing 20-25 kg each were used in this study.
Surgical synovectomy of the left knee, through a medial
arthrotomy, was performed in 10 dogs before the surgical transection of the anterior cruciate ligament (ACL)
of the right knee. The synovial fibroblasts (type B
synoviocytes) isolated from each synovium were used for
autografting (see below). ACL transection of the right
knee was performed through a stab wound on all dogs as
previously described (18). Animals had been anesthetized with pentobarbital (25 mglkg intravenously) and
intubated. Following ACL surgery, the dogs were kept in
animal care facilities within our hospital for 3 days, then
sent to a housing farm where they were left free to
exercise in a large field for 4-6 hours every day.
The animals were divided into 3 experimental
groups, and 48 hours after surgery, the right knee was
injected as follows. Group 1 (controls; n = 7) received 2
ml of phosphate buffered saline (PBS). Group 2 (lac Z ;
n = 5 ) received 2 ml of PBS containing 60 X 106
autologous synovial fibroblasts transduced with the lac Z
gene. Group 3 (IL-1Ra; n = 5 ) received 2 ml of PBS
containing 60 X 10‘ autologous synovial fibroblasts
transduced with the human IL-1 Ra complementary
At 2 weeks and 4 weeks after surgery, synovial
fluid from the right knee of each dog was aspirated
through a medial approach using sterile technique (2-ml
syringe, 21-gauge needle). All dogs were euthanized 4
weeks after ACL surgery.
Cell transduction. Each specimen of synovial
membrane dissected from the dogs’ left knees was
submitted to sequential enzymatic digestion, and the
synovial fibroblasts isolated and cultured as described in
detail elsewhere ( 5 ) . Cells were then transduced with the
appropriate gene as described by Bandara et a1 (19).
Briefly, synovial fibroblasts were grown to -75% confluency in 25-cm2 flasks in Ham’s F-12 medium containing 10% fetal bovine serum. Medium was then removed,
and the cells were infected by adding 1 ml of viral
suspension (titer -106/ml) in the presence of 8 pg/mml of
Polybrene. After a 2-hour incubation, 3 ml of fresh
culture medium was added, and the samples were incubated for another 72 hours. The cells were passaged
once and cultured in 175-cm2flasks until confluency.
Human IL-1Ra cDNA or the marker gene lac Z
was transduced to first passage of cultured synovial
fibroblasts with the retroviral vector MFG-IL-1Ra or
MFG-lac Z (19). Expression of the IL-1Ra and lac Z
genes is regulated by the endogenous 5’ long terminal
repeat (LTR) of the retrovirus. After being infected, the
cells were trypsinized, centrifuged, and resuspended in 2
ml (60 X lo6 cells) of PBS. This cell suspension was
injected intraarticularly directly into the right knee joint
of recipient dogs.
Articular tissue lesion grading. Immediately after killing, the dogs’ right knees were dissected on ice,
and cartilage was examined “blindly” by 2 independent
observers for gross morphologic lesions, as described
previously (13,18). The synovial membrane was dissected and parts were cultured in Dulbecco’s modified
Eagle’s medium for 72 hours, and this conditioned
medium was used to measure IL-1Ra content. Representative specimens were also used for macroscopic and
histologic grading (see below).
The cartilage changes on the medial and lateral
femoral condyles and tibial plateaus were graded separately under a dissecting microscope (Stereozoom;
Bausch & Lomb, Rochester, NY). The depth of erosion
was graded on a scale of 0-4 as follows: grade 0 =
normal surface; grade 1 = minimal fibrillation or a slight
yellowish discoloration of the surface; grade 2 = erosion
extending into superficial or middle layers only; grade
3 = erosion extending into deep layers; and grade 4 =
erosion extending to subchondral bone. The surface area
changes were measured and expressed in mm2.
Histologic cartilage grading. Histologic evaluation was performed on sagittal sections of cartilage from
the lesional areas of each femoral condyle and tibial
plateau (13,18). After dissection, specimens were fixed
in 10% buffered formalin and embedded in paraffin.
Serial sections (5 pm) were prepared and stained with
Safranin 0. The severity of OA lesions was graded on a
scale of 0-14 by 2 independent observers using the
histologic-histochemical scale of Mankin et a1 (20). This
scale evaluates the severity of OA lesions based on the
loss of Safranin 0 staining (scale 0-4), cellular changes
(scale 0-3), invasion of tidemark by blood vessels (scale
0-l), and structural changes (scale 0-6, where 0 =
normal cartilage structure and 6 = erosion of the
cartilage down to the subchondral bone). The scoring
system was based on the most severe histologic changes
in the multiple sections.
Representative specimens of synovial membrane
from the medial and lateral knee compartments were
dissected from underlying tissues. The specimens were
fixed, embedded, and sectioned (5 pm) as above, and
stained with hematoxylin and eosin. For each compartment, 2 synovial membrane specimens were examined
for scoring purposes and the highest score from each
compartment was retained. The average was calculated
and considered as a unit for the whole knee. The severity
of synovitis was graded on a scale of 0-10 (21) by 2
independent observers. adding the scores of 3 histologic
criteria: synovial lining cell hyperplasia (scale 0-2),
villous hyperplasia (scale 0-3), and degree of cellular
infiltration by mononuclear and polymorphonuclear
cells (scale 0-5); 0 indicates normal structure.
IL-1Ra enzyme-linked immunosorbent assay
(ELISA). The level of rHuIL-1Ra in the cell culture
medium, synovial fluid, or synovial membrane explant
conditioned medium of dogs treated with intraarticular
injections of rHuIL-1Ra-transduced cells was determined using a commercial immunoassay kit (Quantikine
Human IL-1Ra; R & D Systems, Minneapolis, MN). For
the synovial fluid, 200 p,I was tested according to the
manufacturer’s instructions. The limit of detection of the
assay is 6.5 pg/ml. The assay used in this study does not
recognize the dog IL-1Ra (unpublished observations).
Statistical analysis. Mean values and SEM were
calculated, and statistical analysis was done using Student’s unpaired t-test.
IL-1Ra levels. The mean t SEM level of IL-1Ra
in the medium from synovial cell culture immediately
after transduction with the IL-1Ra gene was 401.4 t
37.5 ng/106 cells/48 hours (n = 5 ) (Table 1), which was
considerably higher than those obtained with primary
cultures of rabbit or human synovial fibroblasts (unpublished observations). Moreover, this level was comparable to that obtained with cells immediately before the
intraarticular injection (403.4 -+ 2.9 ng/106 cells/48
hours), which was usually about 10 days after transduction. Approximately 1-4 ml of synovial fluid could be
aspirated from 3 dogs at 2 weeks and from 4 dogs at 4
weeks after surgery. The IL-lRa level in the synovial
fluid was 202.8 -+ 131.5 ng/ml2 weeks after surgery and
2.8 5 2.2 ng/ml 4 weeks after surgery, which is considerably diminished. The cultured explants of synovial
Table 1. Amount of IL-1Ra produced by transduced synoviocytes in vitro and in vivo*
Cell culture medium
Freshly transduced
Prior to injection
Synovial fluid
2 weeks
4 weeks
Synovial membrane explants
No. of
lac Z-transduced
Not detectable
Not detectable
401.4 2 37.5 ng/106 cells/48 hours
403.4 ? 2.9 ngilO‘ cellsi48 hours
Not detectable
Not detectable
Not detectable
202.8 2 131.5 ngiml
2.8 2 2.2 ngiml
4.0 2 2.0 ngigm of tissue wet weight
* Osteoarthritis was induced by anterior cruciate ligament transection. Transduced cells were tested for interleukin-1 receptor antagonist (IL-1Ra)
secretion and injected 48 hours after surgery. Synovial fluid was examined at 2 and 4 weeks after surgery. The dogs were killed 4 weeks after surgery,
and synovial membrane explants were tested for the production of IL-1Ra.
membrane also produced a significant amount of IL-1Ra
(4.0 ? 2.0 ng/gm of tissue wet weight).
Macroscopy. Cartilage. In the OA control dogs,
femoral condyles demonstrated small lesions of a mild
grade and size, whereas larger lesions of a slightly more
severe grade were found on the tibial plateaus (Table 2
and Figures 1A and 3).Although slightly smaller lesions
on both condyles and plateaus was noted in the OA dogs
that received the cells transduced with the lac Z gene
than in the OA control dogs, the difference did not reach
statistical significance. However, the grade of both cartilage compartments was similar for OA control and lac
Z groups (Table 2 and Figures 1C and D). Interestingly,
and compared with the lac Z group, the dogs that
received the IL-1Ra gene-transduced cells demonstrated a reduction of cartilage lesions on both the
condyles and the plateaus (Table 2 and Figures 1E and
F). A statistically significant difference was reached for
both the size (P < 0.04) and the grade (P < 0.01) of the
tibial plateau lesions.
Synovial membrane. Synovium from all 3 groups
showed similar changes, with definite signs of inflammaTable 2. Cartilage macroscopic lesions on femoral condyles and
tibial plateaus of a dog model of OA*
Femoral condyles
Tibia1 plateaus
No. of
lac Z
6.4 -C 2.5
2.5 ? 1.6
2.0 t 1.5
0.4 5 0.1
0.5 2 0.3
0.2 2 0.1
13.4 2 4.3
8.0 2 2.9
1.2 2 0.9t
1.0 -C 0.3
1.4 ? 0.3
0.3 + 0.2$
* Osteoarthritis (OA) was induced by anterior cruciate ligament
transection. Transduced cells were injected 48 hours after surgery. The
dogs were killed 4 weeks after surgery. Values are the mean -t SEM.
IL-1Ra = interleukin-1 receptor antagonist.
iP < 0.04 versus lac Z group by Student’s t-test.
$ P < 0.01 versus lac Z group by Student’s &test.
tion. The synovia were hypertrophic and demonstrated a
red-yellowish discoloration with a large number of blood
Microscopy. Cartilage. Specimens from the OA
control (Figures 2A and B) and lac Z (Figures 2C and
D) groups showed a similar degree of morphologic
changes characteristic of early OA. The total histologic
severity of lesions on the condyles and plateaus was
similar in these 2 groups (Figure 3 ) . In the IL-1Ratreated dogs, lesions on condyles and plateaus (Figures
2E and F and Figure 3) were less severe compared with
the lac Z-treated dogs, with statistical significance at
P < 0.06 for the plateaus.
The structural changes (0-6 scale) on the condyles and plateaus were less severe in the IL-1Ra group
(mean ? SEM 0.2 ? 0.1 and 0.5 2 0.2, respectively) than
in the lac Z group (0.8 f 0.5 and 1.4 -t- 0.4). Similarly,
but only for the tibial plateaus, the scores for Safranin 0
staining (0-4 scale) of the plateaus were lower in the
IL-1Ra group (0.9 f 0.2) than in the lac Z group (1.3 ?
0.2). For the cell score (0-3 scale) however, only a trend
toward a reduction could be found in the IL-1Ra group
(0.7 ? 0.1 and 0.5 2 0.1 for condyles and plateaus)
compared with the lac Z group (0.9 ? 0.2 and 1.0 ? 0.2).
Synovial membrane. Synovia from all 3 groups
were thick, with numerous villosities, and showed synovial lining cell hyperplasia (2-5 layers) and marked
infiltration of mononuclear cells. The histologic scores
(0-10 scale) were about equal for the OA control (3.0 -+
0.2, mean ? SEM) and lac Z (2.8 f 0.4) groups, but
higher in the IL-1Ra group (5.5 2 0.6) ( P < 0.008 versus
the lac Z group). Details of the histologic scoring
revealed that the increased inflammation severity in the
IL-1Ra group was due mainly to a more marked cellular
infiltration (0.8 ? 0.3 and 2.7 f 0.4 for the lac Z and
IL-1Ra groups, respectively; P < 0.008).
Figure 1. Macroscopic appearance of cartilage from the femoral
condyles (left panels) and tibial plateaus (right panels). A and B,
Specimens from an osteoarthritic (OA) control dog, treated with
phosphate buffered saline, showing erosion on the condyle and
plateau. C and D, Specimens from a dog that received an intraarticular
injection of synovial cells transduced with the Escherichia coli
/3-galactosidase (lac Z) gene. E and F, Specimens from a dog that
received an intraarticular injection of synovial cells transduced with the
recombinant human interleukin-1 receptor antagonist gene.
vivo delivery, although it is more cumbersome. Successful ex vivo transfer of genes to joints has been reported
for rabbits (17,19,23) and rats (24). Using this approach,
IL-1Ra proved to have an antiarthritic effect in animal
models of RA (25,26), and a clinical trial in humans with
RA is presently under way (27). In OA, however, this is
the first report of the use of gene delivery.
In this study, we demonstrated for the first time
in vivo in an experimental model of OA that synovial
cells transduced with the IL-1Ra gene and grafted
following intraarticular knee injection can retard the
progression of early OA. We showed that the human
IL-1Ra gene was effectively transduced into dog synovial
cells by retroviral infection under the present experimental conditions and that a high level of IL-1Ra was
produced in the transfected knee. In vivo, the level of
synthesis was sustained for about 2 weeks after the
intraarticular injection, and a marked decrease was
noted at 4 weeks. Nevertheless, the autografted synovial
fibroblasts continued to express IL-1Ra for at least 4
weeks, since the synovial explants taken at the time of
Although previous studies have indicated the
usefulness of IL-1Ra as an antiarthritic agent in OA,
there is presently no practical method for delivering this
protein continuously to affected joints. Like all proteins,
IL-1Ra cannot be administered orally, and will be rapidly cleared following intravenous, intramuscular, or
intraarticular injection. Moreover, for the intravenous and
intramuscular routes, only a tiny fraction of the material
will reach the joints, and following systemic administration, it is unlikely that it will target only the OA joints.
Both direct or indirect strategies could be used to
transfer genes to joints (17). Direct in vivo delivery is
clearly more expeditious, but is presently limited by a
lack of suitable vectors (22). Indirect delivery involving
the use of retroviral vectors is highly developed and
commonly used in clinical gene therapy trials. Because
all genetic manipulations occur outside the body and no
infectious agents are introduced into the patient, this
mode of ex vivo gene delivery is currently safer than in
Figure 2. Representative sections of articular cartilage from the femoral condyle (left panels) and tibial plateau (right panels) of (A and B)
an OA control dog treated with phosphate buffered saline, (C and D)
a dog that received an intraarticular injection of synovial cells transduced with the lac Z gene, and (E and F) a dog that received an
intraarticular injection of synovial cells transduced with the recombinant human interleukin-1 receptor antagonist gene. (Safranin 0
stained; original magnification X 100).
Femoral Condyle
5 1
Tibia1 Plateau
Figure 3. Histologic grading of cartilage from femoral condyles and tibial plateaus of osteoarthritic (OA)
dogs. Values are the mean (2SEM) total score (Mankin scale) of lesions from phosphate buffered
saline-treated control dogs or dogs treated with intraarticular injections of synovial cells transduced with
lac Z or interleukin-1 receptor antagonist (IL-1Ra) genes. P versus lac Z group, by Student’s 2-tailed t-test.
killing were producing a measurable level of this
The exact reasons for the marked reduction in
the production of IL-lRa after 2 weeks remain unknown. But this phenomenon was previously reported in
normal rabbit knees into which genetically modified
synovial cells were transplanted by intraarticular injection (19). Two major possibilities could explain this
phenomenon. It could result from a nonimmune mechanism, since as reported (28), the in vivo expression of
genes driven by viral promoters is often of limited
duration. However, the loss of expression of the autografted synovial fibroblasts could reflect either an
immune reaction to specific neoepitopes generated by
trypsinization or a reaction to the human IL-1Ra that
was synthesized in the dog knee.
This hypothesis of an immunologic reaction is
further supported by the data showing that the synovium
from dogs injected with the IL-1Ra-transduced cells had
more inflammation, particularly with regard to the number of mononuclear cells in the sublining tissue. Recent
data from Tripathy et a1 (29) provide strong support for
an immune-based mechanism for the decline of transgene expression in vivo. Such a phenomenon has not
been reported in previous in vivo experiments using the
human IL-1Ra gene (25,26,28) because it has not been
specifically examined. The greater level of inflammation
observed in only the IL-1Ra-transduced group is intriguing also because bacterial P-galactosidase should
have immunogenetic properties in the dog. Although
speculative, it is possible that P-galactosidase is mainly
an intracellular protein, in contrast to IL-lRa, which is
primarily secreted in the extracellular milieu. In support of this hypothesis is the detection of antibodies
against rHuIL-1Ra in dog synovial fluid, in our previous study of this experimental model, in which
intraarticular injections of rHuIL-1Ra were used to
prevent the progression of OA lesions (unpublished
observations) (13).
Injection of synovial cells transduced with the
IL-1Ra gene was shown to be effective in reducing the
progression of cartilage lesions, especially in the tibial
plateaus. In this compartment, and compared with the
lac Z control group, both the macroscopic and microscopic lesions were significantly reduced. The greater
effect of IL-1Ra on plateaus than on condyles relates to
the fact that the plateaus always showed more pronounced lesions (13,18,21) and may have experienced a
greater benefit from the IL-1Ra treatment. Of note,
conditions used in this study represent an extremely
early phase of OA and do not reflect the evolved stage of
the human disease by the time treatment is considered.
Nevertheless, the findings of this study emphasize the
importance of the cytokine IL-1 in this disease process,
at even an early stage. Moreover, these data point to the
importance of the control of IL-1 activity in the treatment of OA and reinforces our in vivo results showing
that intraarticular injection of rHuIL-1Ra in the dog
ACL model could reduce progression of existing lesions
(13) and Baragi and coworkers’ (12) in vitro results
demonstrating that OA cartilage explants transduced
with the IL-1Ra gene can be protected against IL-1induced cartilage degradation. We have recently described an ex vivo strategy for delivering genes to
chondrocytes (30) which will provide an alternative
method for delivering chondroprotective genes to sites
of cartilage loss.
The use of gene therapy for the administration of
IL-1Ra in the treatment of OA is very appealing. The
results from the experimental work of our study and
others (19,22-26) hold interesting promise, even if there
are yet problems to be solved, such as the stability of
gene expression and the choice of vector. The present
data resulting from gene therapy experiments represent
a promising and novel approach to the treatment of OA
and other rheumatic diseases.
1. Pelletier J-P, Martel-Pelletier J, Howell DS: Etiopathogenesis of
osteoarthritis. In, Arthritis and Allied Conditions: A Textbook of
Rheumatology. Edited by DJ McCarty, WJ Koopman. Philadelphia, Lea and Febiger, 1996
2. Pelletier J-P, DiBattista JA, Roughley P, McCollum R, MartelPelletier J: Cytokines and inflammation in cartilage degradation.
Rheum Dis Clin North Am 19:545-568, 1993
3. Okada Y, Takeuchi N, Tomita K, Nakanishi I, Nagase H: Immunolocalization of matrix metalloproteinase 3 (stromelysin) in rheumatoid synovioblasts (B cells): correlation with rheumatoid arthritis. Ann Rheum Dis 48:645-653, 1989
4. Pelletier J-P, McCollum R, Cloutier JM, Martel-Pelletier J: Synthesis of metalloproteases and interleukin 6 (IL-6) in human
osteoarthritic synovial membrane is an IL-1 mediated process.
J Rheumatol 22 (Suppl 43):109-114, 1995
5. Sadouk M, Pelletier J-P, Tardif G, Kiansa K, Cloutier JM,
Martel-Pelletier J: Human synovial fibroblasts coexpress
interleukin-I receptor type I and type I1 mRNA the increased
level of the interleukin-I receptor in osteoarthritic cells is related
to an increased level of the type 1 receptor. Lab Invest 73:347-355,
6. Martel-Pelletier J, McCollum R, DiBattista JA, Faure M-P, Chin
JA, Fournier S, Sarfati M, Pelletier J-P: The interleukin-1 receptor
in normal and osteoarthritic human articular chondrocytes: identification as the type I receptor and analysis of binding kinetics and
biologic function. Arthritis Rheum 35530-540, 1992
7. Firestein GS, Berger AE, Tracey DE, Chosay JG, Chapman DL,
Paine MM, Yu C, Zvaifler NJ: IL-1 receptor antagonist protein
production and gene expression in rheumatoid arthritis and osteoarthritis synovium. J Immunol 149:1054-1062, 1992
8. Pelletier J-P, Mineau F, Ranger P, Tardif G, Martel-Pelletier J: The
increased synthesis of inducible nitric oxide inhibits IL-1Ra synthesis
by human articular chondrocytes: possible role in osteoarthritic
cartilage degradation. Osteoarthritis Cartilage 477-84, 1996
9. ArendWP,bayer J-M: Inhibition of the production and effects of
interleukin-1 and tumor necrosis factor a in rheumatoid arthritis.
Arthritis Rheum 38:151-160, 1995
10. Arner EC, Harris RR, DiMeo TM, Collins RC, Galbraith W:
Interleukin-1 receptor antagonist inhibits proteoglycan breakdown
in antigen induced but not polycation induced arthritis in the
rabbit. J Rheumatol 22:1338-1346, 1995
11. Arend WP: Interleukin-1 receptor antagonist. Adv Immunol 54:
167-227, 1993
12. Baragi VM, Renkiewicz RR, Jordan H, Bonadio J, Harman JW,
Roessler BJ: Transplantation of transduced chondrocytes protects
articular cartilage from interleukin 1-induced extracellular matrix
degradation. J Clin Invest 96:2454-2460, 1995
13. Caron JP, Fernandes JC, Martel-Pelletier J, Tardif G, Mineau F,
Geng C, Pelletier J-P: Chondroprotective effect of intraarticular
injections of interleukin-1 receptor antagonist in experimental
osteoarthritis: suppression of collagenase-l expression. Arthritis
Rheum 39:1535-1544, 1996
14. Campion GV, Lebsack ME, Lookabaugh J, Gordon G, Catalan0
M, and the IL-1Ra Arthritis Study Group: Dose-range and
dose-frequency study of recombinant human interleukin-I receptor antagonist in patients with rheumatoid arthritis. Arthritis
Rheum 39:1092-1101, 1996
15. Evans CH, Robbins PD: Pathways to gene therapy in rheumatoid
arthritis. Curr Opin Rheumatol 8:230-234, 1996
16. Evans CH, Robbins PD: Gene therapy for arthritis. In, Gene
Therapeutics: Methods and Applications of Direct Gene Transfer.
Edited by JA Wolff. Boston, Birkhauser, 1994
17. Bandara G, Robbins PD, Georgescu HI, Mueller GM, Glorioso
JC, Evans CH: Gene transfer to synoviocytes: prospects for gene
treatment for arthritis. DNA Cell Biol 11:227-231, 1992
18. Pelletier J-P, Mineau F, Raynauld J-P, Woessner JF Jr, GunjaSmith Z, Martel-Pelletier J: Intraarticular injections with methylprednisolone acetate reduce osteoarthritic lesions in parallel with
chondrocyte stromelysin synthesis in experimental osteoarthritis.
Arthritis Rheum 37:414-423, 1994
19. Bandara G, Mueller GM, Galea-Lauri J, Tindal MH, Georgescu
HI, Suchanek MK, Hung GL, Glorioso JC, Robbins PD, Evans
CH: Intraarticular expression of biologically active interleukin
1-receptor-antagonist protein by ex vivo gene transfer. Proc Natl
Acad Sci U S A 90:10764-10768, 1993
20. Mankin HJ, Dorfman H, Lippiello L, Zarins A. Biochemical and
metabolic abnormalities in articular cartilage from osteoarthritic
human hips. 11. Correlation of morphology with biochemical and
metabolic data. J Bone Joint Surg Am 53523-537, 1971
21. Pelletier J-P, Martel-Pelletier J, Ghandur-Mnaymneh L, Howell
DS, Woessner J F Jr: Role of synovial membrane inflammation in
cartilage matrix breakdown in the Pond-Nuki dog model of
osteoarthritis. Arthritis Rheum 28554-561, 1985
22. Nita I, Ghivizzani SC, Galea-Lauri J, Bandara G, Georgescu HI,
Robbins PD, Evans CH: Direct gene delivery to synovium: an
evaluation of potential vectors in vitro and in vivo. Arthritis
Rheum 39:820-828, 1996
23. Hung GL, Galea-Lauri J, Mueller GM, Georgescu HI, Larkin LA,
Suchanek MK, Tindal MH, Robbins PD, Evans CH: Suppression
of intraarticular responses to interleukin-I by transfer of the
interleukin-1 receptor antagonist gene to synovium. Gene Ther
24. Makarov SS, Olsen JC, Johnston WN, Schwab JH, Anderle SK,
Brown RR, Haskill JS: Retrovirus mediated in vivo gene transfer
to synovium in bacterial cell wall-induced arthritis in rats. Gene
Ther 2:424-428, 1995
25. Makarov SS, Olsen CC, Johnston WN, Anderle SK, Brown RR,
Baldwin AS, Haskill JS, Schwab JH: Suppression of experimental
arthritis by gene transfer of interleukin-1 receptor antagonist
cDNA. Proc Natl Acad Sci U S A 93:402-406, 1996
26. Otani K, Nita IM, Macaulay W, Georgescu HI, Robbins PD,
Evans CH: Suppression of antigen-induced arthritis in rabbits by
ex vivo gene therapy. J Immunol 156:3558-3562, 1996
27. Evans CH, Robbins PD, Ghivizzani SC, Herndon JK, Kang R,
Tommaino MM, Wasko MC, Watkins SC, Whiteside TK, Glorioso
JC, Lotze MT, Wright TM: Clinical trial to assess the safety,
feasibility and efficacy of transferring a potentially anti-arthritic
cytokine gene to human joints with rheumatoid arthritis. Hum
Gene Ther 7:1261-1280, 1996
28. Palmer TD, Rosman GJ, Osborne WR, Miller AD: Genetically
modified skin fibroblasts persist long after transplantation but
gradually inactivate introduced genes. Proc Natl Acad Sci U S A
88:1330-1334, 1996
29. Tripathy SK, Black HB, Goldwasser E, Leiden JM: Immune
responses to transgene-encoded proteins limit the stability of gene
expression after injection of replication-defective adenovirus vectors. Nat Med 2545-550, 1996
30. Kang R, Marui T, Ghivizzani SC, Nita IM, Georgescu HI, Suh JK,
Robbins PD, Evans CH: Ex vivo gene transfer to chondrocytes in
full-thickness articular cartilage defects: a feasibility study. Osteoarthritis Cartilage 5:139-143, 1997
Без категории
Размер файла
832 Кб
using, experimentov, suppression, antagonisms, osteoarthritis, interleukin, vivo, genes, receptov, therapy, early
Пожаловаться на содержимое документа