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and Suppresses Ongoing Adjuvant Arthritis1
*
Department of Immunology and
Rappaport Family Institute for Research in the Medical Sciences and Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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results in the generation of immunological memory
to its gene product. Upon induction of AA, this memory effectively
inhibited the development of disease. Self-specific Abs developed in
DNA-vaccinated animals were neutralizing in vitro and could adoptively
transfer the beneficial effect of the vaccine. Administration of CFA to
induce a local delayed-type hypersensitivity response rather than AA
did not lead to an elicited production of Abs to the gene product of
the above vaccine. Thus, elicitation of protective immunity is
dependent on the development of an autoimmune condition. Most
importantly, the administration of the TNF- DNA construct after the
onset of disease led to a rapid, long-lasting remission. This suggests
a highly effective way by which a DNA vaccine encoding an autologous
proinflammatory cytokine can be used to reprogram the immune system to
generate protective immunity to its own potentially harmful
activities. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and IL-1, in disease manifestation has
been intensively studied and explored in experimental models that have
been expanded in clinical trails (3, 4, 5, 6, 7, 8) (for general
review, see also Ref. 9). A major disadvantage in treating
long-lasting autoimmune diseases with neutralizing Abs (or soluble
receptor immunotherapy) to proinflammatory mediators lies in their
short t1/2 time, which requires their
repeated administration. We have recently explored a modification of
naked DNA vaccination as a potential way to circumvent this drawback
(10, 11, 12).
We have cloned cDNA encoding various proinflammatory
cytokines/chemokines including TNF-, monocyte chemoattractant
protein-1 (MCP-1), and macrophage-inflammatory protein-1 into a
mammalian vector with a strong viral promoter (CMV) and a recurring
immunostimulatory sequence (13, 14). Upon repeated
administrations, tolerance to the product of each gene of interest was
broken and immunological memory established. Following the initiation
of experimental autoimmune encephalomyelitis (EAE), a T cell-mediated
autoimmune disease of the CNS that serves as an experimental model for
multiple sclerosis, this memory was turned on to provide protective
immunity. Thus, administration of either TNF-,
macrophage-inflammatory protein-1, or MCP-1 DNA vaccines led to a
high state of EAE resistance in Lewis rats that could be adoptively
transferred by the neutralizing Abs generated in response to each gene
product. Surprisingly, elicitation of these Abs was found to be
dependent on the development of the autoimmune condition and regulated
by the immune system in accordance with disease progression (10, 11). This could provide the immune system of a patient with an
autoimmune disease a powerful tool with which to restrain its own
harmful activities. A major disadvantage of exploring the therapeutic
potential of cytokine/chemokine-based DNA vaccines using the EAE model
in the Lewis rat is that both the active and the transferred form of
disease manifest only a transient disease. This model is thus
impractical for the exploration of the effect of such therapy in
chronic or recurrent disease states, particularly when DNA vaccination
was planned for administration after the onset of disease. Under our
working conditions, Lewis rats manifest a long-term AA that includes an
acute phase and a long-lasting (more than 100 days) chronic phase of
disease. The current study explores, for the first time, TNF--based
naked DNA vaccination in AA and reveals its therapeutic potential.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Female Lewis rats, approximately 6 wk old, were purchased from Harlan (Jerusalem, Israel) and maintained under clean conditions in our animal facility.
Immunizations and active disease induction
Rats were immunized s.c. in the base tail with 0.1 ml of CFA (IFA supplemented with 10 mg/ml heat-killed Mycobacterium tuberculosis H37Ra in oil; Difco Laboratories, Detroit, MI). Rats were then monitored for clinical signs daily by an observer blind to the treatment protocol, as we described elsewhere (15). Severity of the disease was quantified subjectively by scoring each limb on a scale of 0–4 to indicate the severity of peripheral joint swelling and erythema: 0 = no signs of disease; 1 = disease evident in a small number of distal joints of the limb; 2 = disease evident in all of distal joints of the limb; 3 = disease evident in all the limb; 4 = severe disease evident in all the limb. The arthritic clinical score was determined as the sum of the scores of all four limbs from each animal (0–16). An observer blind to the experimental protocol quantified the degree of arthritis, indicated by swelling, by measuring front and hind limb circumference using a caliper (Lange Skinfold Caliper; Cambridge Scientific Industries, Cambridge, MA).
DNA vaccination
The sequenced PCR product of rat TNF- (10) was
transferred into a pcDNA3 vector (Invitrogen, San Diego, CA).
Large-scale preparation of plasmid DNA was conducted using Mega prep
(Qiagen, Chatsworth, CA). Cardiotoxin (Sigma, St. Louis, MO) was
injected into the tibialis anterior muscle of 6- to 8-wk-old female
Lewis rats (10 µM per leg). In addition, cDNA encoding rat 
-actin
(the natural cytoplasmic soluble form of -actin) has been obtained
using specific oligonucleotide primers (sense,
5'-ATGGATGACGATATCGCTGCGCTC-3'; antisense,
5'-CTACCGGCCAGCCAGACG-3'). Following cloning and sequence
verification, the above cDNA was ligated into the pcDNA3 vector to be
used as a control DNA vaccine. Five days following the toxin injection,
rats were injected with 100 µg of the pcDNA3 vector with inserts
encoding either TNF- or soluble -actin. Four to five days after
the first immunization, one rat from each group was sacrificed. RT-PCR
was applied on tibialis anterior muscle samples. At that time, muscle
samples form the rat subjected to DNA vaccine encoding TNF-
exhibited an elevated transcription of TNF-, but not of soluble
-actin, and vice versa. Thus, the relevant insert of each inserted
gene is transcribed in the injected muscle. Thereafter, naked DNA
vaccine was administered four times with intervals of 6–7 days between
each injection.
Evaluation of TNF--specific Ab titer in sera of
DNA-vaccinated rats
A direct ELISA assay was used to determine the anti-TNF-
Ab titer in DNA-vaccinated rats. ELISA plates (Nunc, Roskilde, Denmark)
were coated with 50 ng/well commercially available rat rTNF-
(Genzyme, Cambridge, MA). Sera from DNA-vaccinated rats were added in
serial dilutions from 25 to
230 to wells that were, or were not, coated
previously with rTNF-. Calculation of each titer was done by
comparing the OD measured in wells coated with TNF- with those not
coated with this recombinant cytokine. Goat anti-rat
alkaline phosphatase-conjugated IgG Abs (Sigma) were used as a labeled
Ab. p-Nitrophenyl phosphate (Sigma) was used as a soluble
alkaline phosphatase substrate. Results of triplicates were calculated
as log2 Ab titer ± SE.
Purification of Abs
Sera obtained from TNF- DNA-vaccinated rats were subjected to
two steps of purification before being tested for their in vitro
neutralizing characteristics and their ability to affect the course of
AA. At first, a High-Trap Protein G column (Pharmacia, Piscataway, NJ)
was used (according to the manufacturer’s protocol) to purify the IgG
fraction. Then TNF--specific Abs were purified using a
TNF--CNBr-activated Sepharose column, as follows: commercially
available rTNF- (Genzyme, Cambridge, MA) was bound to a
CNBr-activated Sepharose column, according to the manufacturer’s
instructions (Pharmacia Biotech, Uppsala, Sweden; catalog number
17-0820-01). The IgG fraction was then loaded on the column and eluted
by acidic buffer (glycine pH 2.5). Isotype determination (ELISA)
revealed that TNF--specific Abs obtained from DNA-vaccinated rats
are mostly of the IgG2a isotype (data not shown).
Determination of the neutralizing activity of TNF--specific Abs
Determination of the neutralizing activity of TNF--specific
Abs was done as described (16), with the modification of
using the U937 monocyte cell line (CRL-1593.2; American Type Culture
Collection, Manassas, VA), at a concentration of 4 x
104 cells/well, as a target cell for the assay
(10).
Histopathology
Joints were removed at various time points after disease induction, fixed with 10% buffered formalin, decalcified in 5% EDTA in buffered formalin, embedded in paraffin, and sectioned along the midline through the metatarsal region (17). Sections were stained with hematoxylin and eosin and analyzed by a histopathologist blind to the experimental procedure. Evaluation was based upon the level of inflammatory mononuclear cell infiltrate in the synovial membrane, thickness of the synovial lining, joint space narrowing, and periosteal new bone formation. Clinical score was determined as follows: 0 = no evidence of disease; 1 = mild lymphocytic infiltrate; 2 = widespread mononuclear inflammation and thickening of the synovial lining; and 3 = severe bone destruction, new bone formation, and destruction of the synovial lining (17).
Statistical analysis
Significance of differences was examined using Student’s t test. A value of p < 0.05 was considered significant. Mann-Whitney sum of ranks test was used to evaluate significance of differences in mean of maximal clinical score, with p < 0.05 considered significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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naked DNA vaccine
The cloned PCR product of TNF- was ligated into a pcDNA3
mammalian expression vector and used for naked DNA vaccination. Lewis
rats were exposed to four weekly administrations of this construct.
Control rats were injected with either a -actin construct, pcDNA3
vector alone, or PBS. Three weeks after the last immunization, all rats
were immunized with CFA to induce AA. Under our working conditions, AA
manifests a long-lasting form of disease that includes an acute phase,
peaking at about day 20, and a chronic phase that persists for more
than 100 days (Fig. 1
). All control rats
treated with either PBS, pcDNA3 alone, or -actin pcDNA3 (12 per
group) developed a severe form of disease, with a maximal clinical
score (day 20) of 13.5 ± 1.8, 13 ± 1.52, and 13 ±
1.52, respectively (Fig. 1). In contrast, rats subjected to the
administration of the TNF- construct developed a significantly
reduced form of disease (mean maximal score of 6.7 ± 1.1,
p < 0.001 compared with each of the control groups). A
significantly reduced form of disease was also observed in these
animals during the chronic phase of disease (day 45, 1.7 ± 0.7 vs
7.2 ± 0.77, 6.5 ± 1, and 6.7 ± 1.2, p
< 0.001; day 90, 1 ± 0.7 vs 3.3 ± 0.8, 3.3 ± 0.86,
and 3.5 ± 0.7, p < 0.001). Clinical scoring has
also been verified by measuring the changes in the degree of paw
swelling, once again by an observer blind to the experimental
procedure. At all times (days 20, 45, and 90), TNF- DNA-vaccinated
rats exhibited a marked reduction in 
paw swelling compared with
each of the control groups (p < 0.001, data
not shown). Additionally, representative joint sections from all
experimental groups (four animals per group) were obtained on day 30
and screened for histological inflammatory mononuclear cell infiltrate
in the synovial membrane, thickness of the synovial lining, joint space
narrowing, and periosteal new bone formation. Sections obtained from
TNF- DNA-vaccinated rats displayed a marked reduction in each of the
above parameters as compared with control and pcDNA3-treated AA rats
(mean histological score of 12 sections from four animals, 0.5 ±
0.2 in TNF- DNA-vaccinated rats compared with 2.8 ± 0.2,
2.66 ± 0.4, and 3 ± 0 in pcDNA3-, -actin-, or
PBS-treated rats, p < 0.001, respectively). Thus,
TNF--based naked DNA vaccination can serve as a powerful tool to
prevent AA.
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DNA vaccination can potentially elicit both cellular and humoral
responses against products of a given construct (18, 19, 20, 21, 22).
To assess the contribution of the humoral response to the tolerant
state, DNA-vaccinated AA rats were followed for the production of Abs
to TNF-. Then Abs were evaluated for their ability to neutralize
TNF- (in vitro) and to interfere in the development of AA in an
adoptive transfer experiment. Thus, Lewis rats were subjected to
administration of PBS, pcDNA3 alone, -actin construct, or TNF-
naked DNA. Three weeks later, these rats were separated to subgroups
that were immunized with CFA in one of two different ways: 1) footpad
injection to induce a local delayed-type hypersensitivity (DTH)
response, or 2) tail-base administration to induce polyarthritis. At
the peak of the acute phase of disease, the appearance of
anti-TNF- Ab in the serum was determined. Rats developing
polyarthritis develop Ab to TNF- (Fig. 2, A and B), but
not to -actin (data not shown), even without DNA vaccination.
Interestingly, this titer differed significantly not only from the one
in naive rats, but also from that in rats immunized to manifest a local
DTH response (Fig. 2, on day 20 log2 Ab titer of
13 ± 0.55 in AA rats vs 9 ± 0.33 and 8 ± 0.47 in rats
with developing a local DTH response and naive rats, respectively,
p < 0.05 for the comparison of AA rats with each of
the control groups). However, this increase in titer was insufficient
to prevent the development and progression of inflammation in AA (Fig. 1
). In TNF- naked DNA-vaccinated rats, TNF--specific Ab titer was
elicited following AA induction, and to a much lesser extent following
the injection of CFA to induce a DTH response, Fig. 2A
(log2 Ab titer of 25 ± 1.2 vs 16 ±
0.8, p < 0.001). The above elicited Ab titer continued
to persist during the chronic phase of disease (Fig. 2C), as
did the clinical effect of these vaccines (Fig. 1). To determine the
specificity of anti-TNF- Abs, even at the lower dilutions,
rTNF- was added at various concentrations to sera from rats with
developing active AA (day 20, log2 Ab titer of
13 ± 0.3). Then the Ab titer was reassessed. It appears that as
much as 0.1 ng/ml of rTNF- was required to reduce this titer back to
the one observed in control rats immunized with CFA to induce a local
inflammatory process (log2 Ab titer of 9
± 0.3).
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(in vitro)
and transfer AA resistance. IgG from TNF- DNA-vaccinated rats
abolished the cytotoxic activity of TNF- on U937 cells (Fig. 3). (Neutral red uptake as OD at 570 nm
was 0.18 ± 0.007 vs 0.08 ± 0.008, 0.075 ± 0.004, and
0.05 ± 0.008 in the presence of 100 µg/well IgG from TNF-
DNA-vaccinated rats, normal IgG, IgG from pcDNA3-immunized rats, or PBS
and 100 pg/ml TNF-, with backgrounds of 0.17, 0.19, 0.19, and 0.21,
respectively, p < 0.001 for the comparison of the
first group with each of the control groups.) Thus, neutralizing Abs
are produced in naked DNA-vaccinated rats. Purified Abs were then
evaluated for their competence to provide protection from severe
ongoing AA (Fig. 4). Beginning on day 16
after initiation of AA, rats were challenged (i.v.) with 100 µg of
each of these TNF--specific Abs. Control rats were injected with
either PBS, IgG from naive rats, or from AA rats previously
administered with pcDNA3 alone. Repeated administration of
TNF--specific Abs from DNA-vaccinated rats led to a marked reduction
in disease severity as compared with all control groups (day 20, mean
maximal score of 2.25 ± 0.7 compared with 9.5 ± 1.6,
10 ± 1.4, 10 ± 0.7, and 10.5 ± 1.4 in AA rats treated
with either PBS, IgG from pcDNA3-vaccinated rats, or IgG from naive
rats, p < 0.001 for the comparison with each of the
control groups). Clinical scoring has also been quantitated by
measuring limb swelling and verified histologically (not shown). Taken
together, these results may explain, in part, the effect of TNF- DNA
vaccination on disease manifestation. Five to seven days after the last
administration of neutralizing Abs to TNF-, disease severity
regained the level of Ab-untreated AA rats (not shown). This transient
effect of Ab administration further emphasizes the advantageous of
naked DNA vaccination over neutralizing Ab therapy.
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-encoding DNA vaccine
From the practical perspective, we found it highly important to
evaluate the competence of the TNF- naked DNA to interfere with the
progression of an ongoing disease when the vaccine is administered
after the onset of disease. Thus, Lewis rats were immunized with CFA to
induce active AA and divided into four random groups of 12 rats each.
One day after the onset of disease (day 11) and on days 13 and 15, each
group was subjected to administration of either PBS, pcDNA3 alone, a
-actin construct, or the TNF- construct (300 µg per rat). While
all control and pcDNA3-treated rats continued to develop severe AA,
those exposed to the TNF- DNA vaccine exhibited a markedly reduced
disease severity (Fig. 5, day 20, mean
maximal score of 12 ± 0.9 and 10.2 ± 0.62 and 10.5 ±
1.3 in rats treated with either PBS, -actin construct, or pcDNA3
alone vs 6.2 ± 0.76, p < 0.001 for the
comparison of TNF- DNA-vaccinated rats to each control group; day
25, 11.5 ± 1, 11.5 ± 1, and 11.2 ± 0.9 vs 5.3 ±
0.7, respectively, p < 0.001). The marked reduction in
disease severity persisted during the chronic phase of disease (day 60,
4.8 ± 0.8, 5.2 ± 0.8, and 5.5 ± 0.66 vs 1.5 ±
0.7, respectively, p < 0.001). Clinical scores were
confirmed histologically (Fig. 6). Thus,
30 and 60 days after disease induction, representative joint sections
(four rats per group) were obtained from rats that were treated with
the TNF- construct, pcDNA3 alone, or PBS, and evaluated for
histological analysis of synovitis, cartilage loss, and bone erosion,
compared with sections from naive rats. A and B
show naive joints taken with age matching to the experiment rats (day
30); C and D, naive joints that were taken with
age matching to the experiment rats (day 60). E and
F show arthritic joints taken 30 days after disease
induction; G and H, arthritic joints taken 90
days after disease induction. I and J show
pcDNA3-treated joints taken 30 days after disease induction;
K and L show pcDNA3-treated joints taken 60 days
after disease induction. M and N show
TNF--treated joints taken 30 days after disease induction;
O and P show TNF--treated joints taken 60 days
after disease induction. The arrowheads point to the synovial lining
(b = bones, S = synovial membrane). The above representative
sections clearly show that AA rats treated with the TNF- DNA
construct displayed a marked reduction in each of the above parameters
as compared with untreated and pcDNA3-treated AA rats. The beneficial
effect of the treatment was long lasting and covered not only the acute
phase (day 30), but also the chronic phase of disease (day 60). During
the acute phase of disease, sections from PBS- and pcDNA3-treated
control rats displayed a massive inflammatory mononuclear cell
infiltrate in the synovial membrane, an increase in thickness of the
synovial lining, narrowing of the joint space, and notable periosteal
new bone formation (Fig. 6, E and F, and
I and J). At this stage of disease, sections from
TNF- naked DNA-treated rats displayed a substantial reduction in
synovial leukocyte infiltration, synovitis, cartilage loss, and bone
erosion (Fig. 6, M and N). During the chronic
phase of disease, the intensity of the synovial leukocyte infiltration
regressed, while cartilage loss, bone erosion, and periosteal new bone
formation increased (Fig. 6, G and H, and
K and L). Interestingly, and most importantly,
during the chronic phase of disease (day 60), massive cartilage loss,
bone erosion, and periosteal new bone formation that characterized
control and pcDNA3-treated rats were entirely absent in joint sections
of rats treated with TNF- naked DNA vaccine after the induction of
AA (Fig. 6, G and H). Thus, TNF- naked DNA
vaccination can serve as a highly effective treatment for ongoing
arthritis.
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-actin
construct, or our TNF- construct, as described above. Surprisingly,
in all (6/6) TNF- DNA-vaccinated rats, the arthritic score was
markedly reduced within 5–6 days after the DNA vaccine was
administered (Fig. 5B, mean maximal score 2 ± 0 vs
14.1 ± 0.8, p < 0.001). These results are
unexpected, since the process of expressing a transgene and generation
of an immune response against its gene product should have taken much
longer time. A partial explanation for these unexpected, yet highly
interesting observations could be obtained from Fig. 5C. Six
days after administration of each DNA vaccine, sera were obtained from
rats treated with either -actin- or TNF--encoding DNA and
evaluated for TNF-- vs -actin-specific Ab titer, compared with
nontreated (PBS) AA rats. Similar to the results presented in Fig. 3, even without being subjected to TNF--encoding DNA vaccine, AA rats
did mount a significant (p < 0.05) Ab titer to
TNF-, but not to -actin. This titer rapidly accelerated following
the administration of TNF--, but not -actin-encoding DNA vaccine
(Fig. 5C, p < 0.0001). Interestingly,
-actin-encoding DNA vaccine did not elicit anti-self Ab
production within 6 days (Fig. 5C). Taken together, these
results suggest that TNF- DNA vaccines interfere and amplify a
preexisting response. This can provide a partial explanation to the
fast effect of this vaccine, as observed in Fig. 5, A and
B. |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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) are activated
during the course of an autoimmune condition (polyarthritis). The
response they mount is, however, not sufficient to prevent the
development and progression of the autoimmune disease developed in
these animals. Naked DNA vaccination can enhance immunity to these gene
products and thus intervene in disease regulation (Figs. 1, 5, and 6).
Administration of cytokine/hormone-encoding gene in a plasmid (i.m.)
might be used as a way of eliciting the production of its gene product,
which can be used for targeted delivery of gene products
(23, 24, 25), or as a way to elicit an immune response against
the gene product encoded by this construct (10, 11, 26, 27). The mechanistic basis leading to these opposing effects is
not fully understood yet, and may be dependent, in part, by the nature
of the selected plasmid. The existence or absence of repeated
immunostimulatory sequence (CpG) may provide a partial explanation for
the ability of certain plasmids to serve as adjuvants (for a very
recent review, see Gurunathan et al.) (28). In our
experience, administration of inflammatory chemokines and
TNF--encoding constructs ligated into the pcDNA3 plasmid always
resulted in the generation of anti-self immunity that was turned on
during an autoimmune process (10, 11).
The current study shows that either neutralizing Abs (self-specific) or
a targeted DNA vaccine encoding TNF- can markedly reduce,
yet not totally abolish AA (Figs. 1, 4, and 5). On one hand, these
results are remarkable since they imply that targeting the function of
a single proinflammatory cytokine excretes a profound effect on a
disease regulated by various cytokines and chemokines
(29, 30, 31, 32, 33, 34, 35, 36, 37). On the other hand, we (current work) and others
could not use either targeted DNA vaccines, Abs to TNF-, or soluble
TNF- receptor to totally eradicate the disease
(38, 39, 40). The clinical implications of these findings are
highly important since successful clinical trials are now being
conducted using either TNF--specific Abs or TNF- soluble
receptor. One way by which these results could be interpenetrated
relies on the hypothesis that at the clinical readout of disease
resides from a balance between the activity various proinflammatory
cytokines/chemokines such as TNF-, IL-1, RANTES, MCP-1, and
regulatory cytokines such as IL-10 and IL-4. Thus, blockade of the
function of one proinflammatory mediator would significantly alter this
balance, yet this effect is limited since other cytokines can overcome,
in part, this absence. Alternatively, it could also be that
neutralizing TNF- by each of the above means does not totally
abolish TNF- functions.
Neutralizing the activity of TNF- by Abs or even by soluble receptor
therapy as a way to treat arthritis has been explored by several
investigators, and is now in clinical trials. However, a major
disadvantage in treating chronic diseases with xenogenic neutralizing
Abs lies in their immunogenicity. This has motivated investigators to
develop chimeric humanized Abs, and mAbs engineered with human Ig heavy
and light chain yeast artificial chromosome (41). However,
following repeated immunization, these engineered Abs can trigger
allotypic responses. The therapeutic strategy suggested in this study
is of advantage over the above methods since it resulted in the
generation of immunity to autologous Ags that accelerates during the
course of an autoimmune condition in accordance with disease
progression. Even though this makes this type of therapy a very
promising means of treatment for rheumatoid arthritis, and possibly
other T cell-mediated autoimmune diseases (10, 11), one
should also be aware of its limitations. Patients developing a chronic
form of disease will have to spend the rest of their lives with Abs to
their own TNF-. The long-term experience gained in clinical trials
that are now being conducted, in which patients are continually
subjected to soluble TNFR or Abs to TNF-, is highly important for
this perspective. Only if it would appear that long-term neutralization
of TNF- does not lead to harmful side effects should
TNF--encoding DNA vaccine gene therapy be considered. Another
obstacle is the differences in effectiveness of vaccines between animal
models and humans. Yet, many efforts are being spent on improving the
effectiveness of DNA vaccines in humans.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Nathan Karin, Institute of Technology, Bruce Rappaport Faculty of Medicine, Technion, Post Office Box 9649, Haifa 31096, Israel.
3 Abbreviations used in this paper: AA, adjuvant-induced arthritis; DTH, delayed-type hypersensitivity; EAE, experimental autoimmune encephalomyelitis; MCP, monocyte chemoattractant protein.
Received for publication March 20, 2000. Accepted for publication August 14, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, IL-1, and IL-1 receptor antagonist by soluble IL-1 receptors and levels of soluble IL-1 receptors in synovial fluids. J. Immunol. 153:4766.[Abstract]
in rheumatoid arthritis. Arthritis Rheum. 38:151.[Medline]
(cA2) versus placebo in rheumatoid arthritis. Lancet 344:1105.[Medline]
therapy of rheumatoid arthritis. Adv. Immunol. 64:283.[Medline]
) by targeted DNA vaccine confers long lasting resistance to experimental autoimmune encephalomyelitis. Gene Ther. 6:1128.[Medline]
and MCP-1 naked DNA vaccines. J. Autoimmun. 13:21.[Medline]
1 suppresses chronic disease in a streptococcal cell wall-induced arthritis model. J. Clin. Invest. 101:2615.[Medline]
and IL-1 in polymorphonuclear leukocyte and T lymphocyte recruitment to joint inflammation in adjuvant arthritis. Clin. Exp. Immunol. 97:26.[Medline]
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G. Wildbaum, N. Netzer, and N. Karin Plasmid DNA Encoding IFN-{gamma}-Inducible Protein 10 Redirects Antigen-Specific T Cell Polarization and Suppresses Experimental Autoimmune Encephalomyelitis J. Immunol., June 1, 2002; 168(11): 5885 - 5892. [Abstract] [Full Text] [PDF]
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