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Receptor-Deficient Mice1
Laboratories of
*
Immunobiology,
Experimental Chemotherapy, and
Molecular Immunology, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Leuven, Belgium; and
AnorMed, Langley, British Columbia, Canada
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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R-deficient
DBA/1 mice was shown to be reduced in severity by treatment with the
bicyclam derivative AMD3100, a specific antagonist of the interaction
between the chemokine stromal cell-derived factor-1 (SDF-1) and its
receptor CXCR4. The beneficial effect of the CXCR4 antagonist was
demonstrable when treatment was initiated between the time of
immunization and appearance of the first symptoms. Treatment also
reduced the delayed-type hypersensitivity response to the autoantigen,
collagen type II. These observations are indicative of an action on a
late event in the pathogenesis, such as chemokine-mediated attraction
of leukocytes toward joint tissues. The notion of SDF-1 involvement was
further supported by the observation that exogenous SDF-1 injected in
periarthritic tissue elicited an inflammatory response that could be
inhibited by AMD3100. The majority of leukocytes harvested from
inflamed joints of mice with CIA were found to be Mac-1+
and CXCR4+, and AMD3100 was demonstrated to interfere
specifically with chemotaxis and Ca2+ mobilization induced
in vitro by SDF-1 on Mac-1+/CXCR4+ splenocytes.
We conclude that SDF-1 plays a central role in the pathogenesis of
murine CIA, by attracting Mac-1+/CXCR4+ cells
to the inflamed joints. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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) first described for
its potent activity against HIV infection (7, 8) and
presently under investigation for clinical applicability in AIDS
patients (9). It selectively antagonizes the CXCR4
(10, 11), which acts as a coreceptor for HIV
(12). AMD3100 also inhibits the in vitro chemotactic and
intracellular Ca2+ flux responses of human
monocytes to SDF-1 (10). This makes AMD3100 an ideal tool
to evaluate the physiological and pathological importance of
SDF-1/CXCR4 interactions. However, AMD3100 is rapidly cleared from the
circulation, and treatment schedules in mice require the use of osmotic
minipumps (13). This restricts the time of treatment to a
maximum of 2 wk. Because development of CIA in wild-type DBA/1 mice
takes on average 6 wk (14), we used IFN-R-deficient
(IFN-R knockout (KO)) mice, as these are known to develop joint
lesions that are pathologically identical with those in wild-type mice
in much less time (average 3 wk) (14, 15, 16).
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Chicken collagen type II (CII; EPC, Owensville, MO) was
dissolved in 0.05 M acetic acid at 2 mg/ml and emulsified in an equal
volume of CFA supplemented with 1.5 mg/ml heat-killed
Mycobacterium butyricum (Difco, Detroit, MI). Eight- to
12-wk-old IFN-R KO mice of DBA/1 strain (17) were
sensitized with a single 100-µl intradermal injection of the emulsion
at the base of the tail. Mice were examined daily for signs of
arthritis. The disease severity was recorded for each limb, as follows:
score 0, normal; 1, redness and/or swelling in one joint; 2, redness
and/or swelling in more than one joint; 3, redness and/or swelling in
the entire paw; 4, deformity and/or ankylosis. In each experiment, mice
were age and sex matched in the different groups.
Histology
Fore- and hind limbs (ankles and interphalanges) were fixed in 10% Formalin and decalcified with formic acid. Paraffin sections (4 µm) were stained with H&E. Severity of arthritis was evaluated blindly, using three parameters: infiltration of mono- and polymorphonuclear cells, hyperplasia of the synovium, and pannus formation. Each parameter was scored on a scale from 0 to 3: score 0 (absent), score 1 (weak), score 2 (moderate), score 3 (severe).
Treatments with AMD3100
AMD3100 treatment schedules were virtually identical with those
developed on the basis of careful pharmacokinetic evaluation by Datema
et al. (13). We used the same Alzet osmotic minipumps
(Alza, Palo Alto, CA), implanted dorsolaterally under the skin. All
mice were anesthetized with 200 µl of a solution in PBS of 0.2%
Rompun (Bayer, Brussels, Belgium) and 1% Ketalar (Parke-Davis,
Zaventem, Belgium). In most experiments, number 2002 pumps (delivering
0.5 µl/h for 14 days) were used and were filled with either 3 or 10
mg of AMD3100 in 200 µl of PBS (AMD3100 pump, delivering the drug at
a rate of, respectively, 180 or 600 µg/day). According to Datema et
al. (13), this corresponds to steady serum levels of,
respectively, 0.3 or 1 µg/ml. In experiment 2 of Fig. 4B,
number 1007D pumps were used and delivered AMD3100 at a rate of 576
µg/day for 7 days. In two experiments (Fig. 2, A and D), groups
of mice implanted with pumps containing PBS only (without AMD3100) were
included. All other untreated mice were anesthetized like treated ones,
but were not implanted with pumps.
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Individual sera were tested for the amount of Abs directed to chicken CII by ELISA, as described (14). For evaluation of DTH reactivity, mice were challenged in the right footpad with 10 µg of chicken CII in 20 µl of PBS. The left footpad received 20 µl of PBS. DTH response was measured as percentage of swelling at 24 and 48 h postchallenge.
Flow cytometric analysis
Spleens were diced and passed through cell strainers (BD
Labware, Franklin Lakes, NJ). Blood, taken by heart puncture, was
collected on heparin. Cells from joint cavities were obtained from an
opening that was made with a 25-gauge needle in the skin on the dorsal
side of the foot, just distally from the interphalangeal space. Eighty
microliters of cold PBS was injected in the interphalangeal spaces, the
needle being inserted on the dorsal side of the foot and oriented from
proximal to distal. Fluid exiting spontaneously from the distal opening
was collected and was only used when it was found to contain <10% of
RBCs. Erythrocytes were removed from blood and splenocyte suspensions
by lysis with NH4Cl (0.83% in 0.01 M Tris-HCl,
pH 7.2; two consecutive incubations of 5 and 3 min, 37°C). Remaining
cells were washed, resuspended in cold PBS, and counted. Aliquots of
2 x 105 cells in 0.2 ml were preincubated
(30 min) with a FcR-blocking Ab (2.4G2, 1 µg/ml; BD PharMingen, San
Diego, CA) and then stained for 30 min with FITC-conjugated
anti-Mac-1 (CD11b) Ab and PE-conjugated anti-CXCR4 (12G5; R&D
Systems Europe, Abingdon, U.K.). Simultest control
1/2a (BD Biosciences,
San Jose, CA) was used as isotype control. Cells were analyzed by a
FACScan flow cytometer (BD Biosciences).
Measurement of intracellular calcium flux
An enriched Mac-1+ cell population was obtained by depletion of T and B cells from splenocytes using CD90 (Thy-1.2) and CD45R (B220) microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany); the purity was analyzed by flow cytometry. Mac-1+ cells were loaded with the fluorescent calcium indicator Fluo-3 acetoxymethyl (Molecular Probes, Leiden, The Netherlands) at 4 µM for 45 min at room temperature. After thorough washing with HBSS containing 20 mM HEPES and 0.2% BSA (pH 7.4), the cells were resuspended in the same buffer and were seeded at 3 x 105 cells/well into a 96-well plate, containing AMD3100 at different concentrations. After preincubation of the cells for 20 min in the presence of AMD3100, 50 ng/ml murine SDF-1 (R&D Systems) was added, and the fluorescence in function of time was monitored simultaneously in all wells by a Fluorometric Imaging Plate Reader (Molecular Devices, Sunnyvale, CA).
Chemotactic assay
SDF-1-induced cell migration was assessed using 5-µm-pore Transwell filter membranes (Costar, Boston, MA). The membrane inserts were placed in the wells of a 24-well plate, containing 600 µl of HBSS (see above) with murine SDF-1. After preincubation with AMD3100 at different concentrations, 1 x 106 purified Mac-1+ cells in 100 µl of buffer were loaded into each Transwell filter. The plate was then incubated at 37°C for 3.5 h, whereafter the filter inserts were carefully removed and the migrated cells were collected from the wells and counted in a flow cytometer. Chemotactic index is the number of migrated cells obtained with 0.1 µg/ml SDF-1 divided by the number of migrated cells in the negative control (without SDF-1). Migrated cells were characterized after centrifugation by cytospin (Shandon, Cheshire, U.K.) and staining with Hemacolor (Merck, Darmstadt, Germany).
Semiquantitative analysis of SDF-1 mRNA by RT-PCR
The skin was removed from mouse hind limbs, and ankle joints
were dissected. Joint tissues were snap frozen in liquid nitrogen and
pulverized. RNA was extracted by the TRIzol Reagent method (Life
Technologies, Gaithersburg, MD). First-strand cDNA was synthesized
using 1 µg of total RNA and 4.5 U of RAV-2 reverse transcriptase
(Amersham, Aylesbury, U.K.). The reaction mixture was incubated for 80
min at 42°C, followed by a denaturation step of 5 min at 95°C. As
an external control for the amount of RNA in the different samples,
2-microglobulin (2m)
mRNA was first quantitated by PCR with the primers
2mFOR (5'-CTGACCGGCCTGTATGCTATCC-3') and
2mBACK (5'-CATGTCTCGATCCCAGTAGACGG-3')
using 0.25 U SuperTaq DNA polymerase (HT Biotechnology, Cambridge,
U.K.). After determining the volumes of first-strand cDNA sample that
gave the same yield of 2m PCR product, SDF-1
mRNA was quantified with SDF-1-specific primers SDF-1FOR
(5'-TCAGCCTGAGCTACCGATGC-3') and SDF-1BACK
(5'-ACGGATGTCAGCCTTCCTCG-3'). All PCR were performed in 50-µl
reaction mixtures on a GeneAmp PCR System 2400 (PerkinElmer,
Norwalk, CT) for 30 cycles (2m and SDF-1), in
which each cycle consists of 30 s at 95°C, 30 s at 64°C,
and 30 s at 72°C, with a final extension step of 5 min at
72°C. Equal amounts of PCR products (for 2m
and SDF-1) of the samples were analyzed on 1.5% agarose gels and
stained with ethidium bromide.
In vivo effects of SDF-1
Murine SDF-1 (1 µg in 20 µl of PBS) was injected in the right footpads of immunized mice. Redness and swelling reaction were recorded at different time points. The swelling reaction was measured by calipers and expressed as the mean percentage of increase in footpad thickness relative to the thickness before the SDF-1 challenge. At 24 or 48 h postchallenge, cells from the inflamed area were obtained in a similar way as described for arthritic joints, and were stained with Hemacolor after centrifugation by cytospin.
Statistical analyses
The Mann-Whitney U test, Student’s t test, and the log rank test for survival curves (referred to in Ref. 14) were used as indicated.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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R KO mice were implanted with
osmotic minipumps releasing AMD3100 at a rate of 180 µg/day for 14
days (see Materials and Methods). The treatment regime was
adapted from experiments in which AMD3100 successfully inhibited HIV
infection in SCID-hu Thy/Liv mice (13). Two additional
groups of control mice, either implanted with pumps containing PBS or
not implanted, were included. One day after pump implantation, all mice
were immunized with CII in CFA, and examined daily for clinical signs
of arthritis. In control mice and in mice receiving PBS, the symptoms
of arthritis started to appear as early as day 19 (Fig. 2
A).
In AMD3100-treated mice, the first symptoms appeared about 2 wk later.
By that time, nearly 50% of the control and PBS-treated mice had
developed arthritis. The mean day of disease onset in AMD3100-treated
mice was significantly different from that in control or PBS-treated
mice (p < 0.05, Mann-Whitney U
test); mean arthritis scores of AMD3100 and PBS-treated mice were
significantly different (p < 0.05) in the time
interval from day 28 to day 37. The differences in disease onset times
and scores of arthritis were confirmed in an additional experiment
(Fig. 2B). The clinical scores of arthritis were
substantially lower in AMD3100-treated mice (Fig. 2B,
p < 0.03 from day 19 to day 40, Mann-Whitney
U test).
The effect of treatment with AMD3100, initiated at a later time point,
was also investigated. In two independent experiments, mice were
implanted with AMD3100 pumps on day 7 postimmunization. Here again, the
incidence of arthritis was lower, the onset was delayed, and the
clinical disease scores were significantly lower in mice treated with
AMD3100 (Fig. 2, C and D). These experiments were
terminated on days 25 and 20 to allow for, respectively, histological
examination and testing DTH (see below). The joints were analyzed
histologically at this rather early time point, to ascertain that the
protective effect of AMD3100 on the clinical symptoms of arthritis
correlated with inhibition of tissue damage. The data, summarized in
the inset of Fig. 2C, confirmed the protective
effect of AMD3100 against the development of arthritis: infiltration of
mono- and polymorphonuclear cells, hyperplasia of the synovium, and
pannus formation were all dramatically reduced in treated
mice.
Finally, we assessed whether AMD3100 could control CIA when
administered at the time of disease onset. To this end, CII-immunized
mice were divided into two groups at the time when clinical symptoms of
arthritis started to appear. In one group, the mice were implanted with
minipumps containing AMD3100, whereas in the other group the mice were
left untreated. In a first experiment, in which mice were treated with
the regular dose of AMD3100 (180 µg/day for 14 days), the average
clinical score did remain below that in the control group, although the
difference was not statistically significant (mean end scores ±
SEM on day 40 postimmunization: 0.6 ± 0.3 for 10 AMD3100-treated
mice vs 2.3 ± 0.9 for 10 control mice). In a second experiment,
the dose of AMD3100 was increased to 600 µg/day for 14 days (see
Materials and Methods) and pumps were implanted at the time
of disease onset, i.e., on day 22 postimmunization. As can be seen in
Fig. 2E, the clinical scores in treated mice were
significantly lower than in control mice. Whereas control mice showed
scores gradually increasing to a maximum averaging 5.5, treated mice
remained below score 1 throughout the experiment.
We also measured body weight changes to evaluate potential toxicity of
the compound. In fact, AMD3100-treated mice gained more weight than the
controls (data not shown), suggesting that reduced CIA scores were not
due to possible toxic effects of the compound (18).
Moreover, spleens, lymph nodes, livers, kidneys, lungs, hearts, and
intestines of AMD3100-treated mice had normal histological appearance
(data not shown). The protective effect of AMD3100 on the clinical
severity of arthritis was confirmed by histological examination (day
53). Representative images are shown in Fig. 3. Treated mice showed a marked decrease
in cellular infiltration and hyperplasia of the synovium. The mean
joint scores of AMD3100-treated vs control mice (n = 16
limbs from four mice) were, respectively, 0.25 ± 0.12 vs
2.06 ± 0.17 for infiltration (p <
0.00001, Student’s t test) and 0.63 ± 0.18 vs
2.25 ± 0.19 for hyperplasia (p <
0.0001). Moreover, a moderate to severe pannus formation, which is
often seen at this stage of the disease (Fig. 3C), was
barely detectable in AMD3100-treated mice (6% in limbs of treated mice
vs 81% in those of control mice; mean scores were, respectively,
0.50 ± 0.16 vs 2.19 ± 0.19, p <
0.00001).
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A. Serum levels of
anti-CII Abs (Fig. 4A)
were not affected by AMD3100 treatment. The cellular immune response to
CII was examined by measuring DTH reactivity against CII injected in
the footpad. On day 7 postimmunization with CII in CFA, treatment with
either AMD3100 or PBS was initiated. On day 20, no clinical signs of
arthritis were detectable in the AMD3100-treated mice, whereas by that
time 50% of the mice in the PBS group had developed limb inflammation.
On this day, mice without clinical swelling of the hind limbs were
selected from each group and challenged with CII by injection in the
footpad. As shown in Fig. 4B (Expt. 1), in PBS pump-treated
mice, a distinct DTH reaction was observed at 24 and 48 h
postchallenge. The swelling reaction was significantly less pronounced
in the mice that had received AMD3100. Importantly, a reduced swelling
reaction was also seen in another experiment in which AMD3100 treatment
was started as late as day 25 postimmunization, i.e., 2 days before the
challenge with CII (Fig. 4, B (Expt. 2) and
C).
As the anti-HIV effect of AMD3100 originates from specific binding
to the chemokine receptor CXCR4 (10, 11) (vide infra), the
inhibitory effect of AMD3100 on CIA and on the DTH reaction against CII
may be due to interference with migration of
CXCR4+ leukocytes into the inflamed tissues.
Therefore, we examined the expression of CXCR4 on splenocytes, on blood
leukocytes, and on cells obtained from the arthritic joints of mice
with CIA. As shown in Fig. 5, all three
cell populations strongly expressed CXCR4. In accordance with the
established involvement of Mac-1+
(CD11b+) cells in the pathogenesis of CIA
(16, 19), >90% of the cells harvested from arthritic
joint cavities appeared to be Mac-1+.
Interestingly, a pattern of CXCR4 and Mac-1 expression similar to that
in the arthritic joints was seen in inflamed footpads after a CII
challenge to elicit DTH.
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A). Then, we also
investigated the capacity of AMD3100 to inhibit SDF-1-induced migration
of Mac-1+ cells (Fig. 6B). Clearly,
murine SDF-1 elicited a strong chemotactic response on
Mac-1+ cells: at 0.1 and 0.5 µg/ml, SDF-1
triggered migration of, respectively, 14 and 30% of the cells, with a
chemotactic index of 30 (for 0.1 µg/ml SDF-1). Moreover, the
percentage of cells that migrated in response to SDF-1 gradually
decreased when the cells were preincubated with AMD3100 at increasing
concentrations (Fig. 6B). Further characterization of the
cells that were attracted by SDF-1 revealed that >95% stained
positively for both Mac-1 and CXCR4. As shown in Fig. 6C,
the vast majority of migrated cells contained a ring-shaped nucleus, a
characteristic attributed to immature monocytes and neutrophils
(20). It is important to note that cells of a similar
phenotype are present in the synovia of arthritic joints (Fig. 3C).
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). Expression was present in joint
tissues of mice free of symptoms (either naive or immunized mice), but
was more pronounced in inflamed joints. We next tested in vivo
interference of AMD3100 with SDF-1-induced cell migration in a
footpad-swelling test. Preliminary tests had established that local
injection of murine SDF-1 (doses of 1, 5, or 7.5 µg) in the footpad
induces redness and swelling in mice that had previously been immunized
with CII in CFA. Therefore, on day 19 postimmunization, mice were given
an i.p. injection of AMD3100 (25 µg in 100 µl of PBS), followed by
injection of 1 µg of SDF-1 in the right footpad; controls received
PBS without the drug and were similarly challenged with SDF-1.
Increases of footpad thickness (mean percentage ± SEM) were most
pronounced at 4 h after SDF-1 injection and were 9.1 ± 3.7
in treated vs 36.8 ± 6.5 in controls (n = 4;
p < 0.05 by Mann-Whitney U test). Fluid
aspirated from the footpad of controls contained leukocytes (mainly
polymorphonuclear cells), whereas virtually no cells could be obtained
from the footpads of AMD3100-treated mice.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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SDF-1 mRNA, unlike that of most other chemokines, is constitutively expressed in a wide range of tissues (2). Overexpression has recently been reported to occur in cultured synoviocytes of rheumatoid arthritis patients, but, interestingly, not in those of patients with osteoarthritis, who lack hyperplasia and inflammatory cell infiltration. In our study, we found expression of SDF-1 mRNA in joint tissue extracts of mice with signs of arthritis. This clearly suggests a role for SDF-1 in the invasion of leukocytes into the inflamed joint tissues (5, 6, 21). These findings are in line with our observation that, in mice sensitized to develop CIA, injection of SDF-1 in the footpad resulted in an inflammatory swelling reaction that was inhibited by AMD3100.
SDF-1 was originally cloned from a bone marrow stromal cell line (22) and found to act as a B cell growth factor (23). It was later found also to induce chemotaxis of immunocompetent (3) and progenitor cells (24) and to be involved in myelopoiesis (25, 26). However, we found no evidence to support the concept that in the CIA model, AMD3100 interfered with myelopoiesis and B cell growth. First, flow cytometric analysis evaluating the following markers (CD11b, CD19, B220, CD4, and CD8) did not reveal effects of AMD3100 on the proportions of Mac-1+, B, or T cell populations in spleen, blood, and bone marrow. In addition, cytospin preparations of spleen and bone marrow cell suspensions failed to reveal any effect of the drug on the numbers of immature myeloid cells (data not shown).
We also assessed whether SDF-1 and AMD3100 could interfere with
osteoclastogenesis, which was recently reported to represent an
important event in the pathogenesis of adjuvant-induced joint
destruction (27). AMD3100 (20 µg/ml) failed to affect
osteoclast formation induced in bone marrow and spleen cultures by
stimulation with either osteoclast differentiation factor or TNF-
.
In this setting, SDF-1 was unable to induce the formation of
tartrate-resistant acid phosphatase-positive osteoclast-like cells
(data not shown); therefore, it seems unlikely that SDF-1 or CXCR4 is
involved in osteoclastogenesis.
Our observations provide evidence for a proinflammatory role of SDF-1
in vivo, and suggest that AMD3100 or similar specific inhibitors of
SDF-1/CXCR4 may be useful not only for the treatment of rheumatoid
arthritis, but also for the treatment of certain DTH-mediated
allergies. However, before tests in patients can be considered,
evidence for activity in animals with an intact IFN- system is
needed, and strict absence of toxic side effects should be
demonstrated.
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Acknowledgments |
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R KO mice. |
Footnotes |
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2 Address correspondence and reprint requests to Dr. Patrick Matthys, Rega Institute, University of Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium. E-mail address: Patrick.Matthys{at}rega.kuleuven.ac.be
3 Abbreviations used in this paper: CIA, collagen-induced arthritis; 2m, 2-microglobulin; CII, collagen type II; DTH, delayed-type hypersensitivity; KO, knockout; SDF-1, stromal cell-derived factor-1.
Received for publication May 11, 2001. Accepted for publication August 2, 2001.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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L. Zou, B. Barnett, H. Safah, V. F. LaRussa, M. Evdemon-Hogan, P. Mottram, S. Wei, O. David, T. J. Curiel, and W. Zou Bone Marrow Is a Reservoir for CD4+CD25+ Regulatory T Cells that Traffic through CXCL12/CXCR4 Signals Cancer Res., November 15, 2004; 64(22): 8451 - 8455. [Abstract] [Full Text] [PDF]
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R. Lande, E. Giacomini, B. Serafini, B. Rosicarelli, G. D. Sebastiani, G. Minisola, U. Tarantino, V. Riccieri, G. Valesini, and E. M. Coccia Characterization and Recruitment of Plasmacytoid Dendritic Cells in Synovial Fluid and Tissue of Patients with Chronic Inflammatory Arthritis J. Immunol., August 15, 2004; 173(4): 2815 - 2824. [Abstract] [Full Text] [PDF]
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M. Lenoir, B. Djerdjouri, and A. Perianin Stroma Cell-Derived Factor 1{alpha} Mediates Desensitization of Human Neutrophil Respiratory Burst in Synovial Fluid from Rheumatoid Arthritic Patients J. Immunol., June 1, 2004; 172(11): 7136 - 7143. [Abstract] [Full Text] [PDF]
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S. J. Curnow, K. Wloka, J. M. Faint, N. Amft, C. M. G. Cheung, V. Savant, J. Lord, A. N. Akbar, C. D. Buckley, P. I. Murray, et al. Topical Glucocorticoid Therapy Directly Induces Up-Regulation of Functional CXCR4 on Primed T Lymphocytes in the Aqueous Humor of Patients with Uveitis J. Immunol., June 1, 2004; 172(11): 7154 - 7161. [Abstract] [Full Text] [PDF]
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H. Bruhl, J. Cihak, M. A. Schneider, J. Plachy, T. Rupp, I. Wenzel, M. Shakarami, S. Milz, J. W. Ellwart, M. Stangassinger, et al. Dual Role of CCR2 during Initiation and Progression of Collagen-Induced Arthritis: Evidence for Regulatory Activity of CCR2+ T Cells J. Immunol., January 15, 2004; 172(2): 890 - 898. [Abstract] [Full Text] [PDF]
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C-Q Chu, Z Song, L Mayton, B Wu, and P H Wooley IFN{gamma} deficient C57BL/6 (H-2b) mice develop collagen induced arthritis with predominant usage of T cell receptor V{beta}6 and V{beta}8 in arthritic joints Ann Rheum Dis, October 1, 2003; 62(10): 983 - 990. [Abstract] [Full Text]
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K. Balabanian, J. Couderc, L. Bouchet-Delbos, A. Amara, D. Berrebi, A. Foussat, F. Baleux, A. Portier, I. Durand-Gasselin, R. L. Coffman, et al. Role of the Chemokine Stromal Cell-Derived Factor 1 in Autoantibody Production and Nephritis in Murine Lupus J. Immunol., March 15, 2003; 170(6): 3392 - 3400. [Abstract] [Full Text] [PDF]
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J. L. Pablos, B. Santiago, M. Galindo, C. Torres, M. T. Brehmer, F. J. Blanco, and F. J. Garcia-Lazaro Synoviocyte-Derived CXCL12 Is Displayed on Endothelium and Induces Angiogenesis in Rheumatoid Arthritis J. Immunol., February 15, 2003; 170(4): 2147 - 2152. [Abstract] [Full Text] [PDF]
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T. Murakami, W. Maki, A. R. Cardones, H. Fang, A. Tun Kyi, F. O. Nestle, and S. T. Hwang Expression of CXC Chemokine Receptor-4 Enhances the Pulmonary Metastatic Potential of Murine B16 Melanoma Cells Cancer Res., December 1, 2002; 62(24): 7328 - 7334. [Abstract] [Full Text] [PDF]
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C. J. Scotton, J. L. Wilson, K. Scott, G. Stamp, G. D. Wilbanks, S. Fricker, G. Bridger, and F. R. Balkwill Multiple Actions of the Chemokine CXCL12 on Epithelial Tumor Cells in Human Ovarian Cancer Cancer Res., October 15, 2002; 62(20): 5930 - 5938. [Abstract] [Full Text] [PDF]
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N. W. Lukacs, A. Berlin, D. Schols, R. T. Skerlj, and G. J. Bridger AMD3100, a CxCR4 Antagonist, Attenuates Allergic Lung Inflammation and Airway Hyperreactivity Am. J. Pathol., April 1, 2002; 160(4): 1353 - 1360. [Abstract] [Full Text] [PDF]
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2 values of 2.8 and
4.4 for the two independent experiments, A and
B, respectively, yielding an overall p
value < 0.05. C, Two groups of mice were either
left untreated (control, n = 7) or implanted with
AMD3100-containing pumps (n = 8) on day 7. The
graph shows mean disease scores ± SEM (significantly different
from day 21 onward; p < 0.02, Mann-Whitney
U test). Inset, Six mice of each group
(including the only two mice with clinical symptoms in the AMD3100
group) were sacrificed on day 25. The forepaws, toes, and ankle
sections were scored histologically for three parameters of arthritis.
Bars represent averages ± SEM of at least 27 sections of six
mice. *, p < 0.05 (infiltration),
p < 0.0001 (hyperplasia), p <
0.02 (pannus); Student’s t test. D, Two
groups of six mice were treated with AMD3100- or PBS-containing pumps
on day 7 postimmunization with CII. On day 20, clinical symptoms of
arthritis were observed in three control mice, but not in animals
treated with AMD3100. On that day, mice were subjected to DTH testing,
as further explained in Fig. 4
