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Vol. 283, Issue 1, 123-130, 1997
Department of Biochemical Pharmacology, The William Harvey Research Institute, London, United Kingdom (S.J.G., R.J.F., M.A.M., M.P.); Istituto di Farmacologia e Farmacognosia, Facolt[aa]a di Farmacia, Palermo, Italy (L.P.); Rheumatic Disease Unit, Faculty of Medicine, Université de Sherbrooke, Sherbrooke, Canada (R.d.M., A.L.); Department of Inflammation/Autoimmune Diseases, Hoffman-La Roche Inc., Nutley, New Jersey (B.A.W.)
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Abstract |
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 Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Injection of monosodium urate (MSU) crystals, the etiological cause of gouty arthritis, into murine peritoneal cavities produced an intense recruitment of polymorphonuclear leukocytes (PMN). After 3 mg MSU crystal injection, cell influx was maximal (~ 10 × 106 cells per mouse) at 6 hr postinjection and sustained up to the 24 hr time-point. In mice depleted of mast cells by administration of compound 48/80 72 hr before challenge with MSU crystals a lower PMN influx was measured (58% reduction). The occurrence of endogenous mast cell activation, in the MSU response, was validated by the observation that MSU challenge reduced by more than 90% the number of intact mast cells recovered in the peritoneal washes. Pretreatment of mice with a histamine H1 antagonist (tripolidine; 0.5 mg/kg) or a platelet-activating factor receptor antagonist (WEB2086; 10 mg/kg) significantly reduced by 50 to 60% the number of PMN recovered from the peritoneal cavities. The molecular determinants of this process of leukocyte recruitment were also investigated. Treatment of mice with an anti-CD62P or anti-CD62E monoclonal antibody (mAb; 100 µg i.v.) produced a distinct inhibition of PMN recruitment measured at 6 hr, whereas only a combined administration of both monoclonal antibodies was effective in reducing by 60% the influx of PMN caused by the MSU crystals within 24 hr. In conclusion, these data highlight a role for endogenous mast cells and for endothelial-derived selectins in MSU crystal-induced PMN recruitment into the peritoneal cavity, and may be useful to dissect molecular mechanism(s) which may be operating in gouty arthritis.
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Introduction |
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Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Deposition
of MSU and CPPD crystals in the joint articular space is the
etiological cause of acute inflammatory conditions such as gout and
pseudogout, respectively (McCarthy et al., 1962
; Dieppe
et al., 1979). Clinically, these inflammatory diseases are
associated with edema and erythema of the joints with consequent severe
pain. A strong infiltration of leukocytes in the intraarticular and
periarticular space is also characteristic of these pathologies. In
particular, PMN are the predominant cell type recovered from these
inflammatory joints (Terkeltaub, 1992; Dieppe et al., 1979).
The clear relationship between MSU crystals and gouty arthritis has, in
the past, prompted the characterization of experimental models of
crystal-induced inflammation. For instance, injection of crystals into
a preformed rat air-pouch (Brooks et al., 1987) or within
the rat pleural cavity (Sedgwick et al., 1985) produced an
intense PMN accumulation associated with generation of chemoattractants such as leukotriene B4 (Brooks et al.,
1987). More recently, the ability of MSU crystals to activate human
neutrophils in vitro has been reported. Incubation of these
cells with tumor necrosis factor produces release of both interleukin-1
and interleukin-1 receptor antagonist, and coaddition of MSU crystals
potentiate the release of the former without affecting the latter
(Roberge et al., 1994). Furthermore, in similar conditions
the synthesis of a CXC chemokine selective for PMN, such as
interleukin-8, has also been observed, but not that of a CC chemokine
selective for monocytes, such as monocyte chemoattractant protein-1
(Hachica et al., 1995). In addition, MSU crystal activation
of mononuclear phagocytes, which are normally found in the joint space,
also induces secretion of interleukin-8 (Terkeltaub et al.,
1991). All these studies provide further evidence that gout is mainly a
PMN driven pathology.
The events that regulate PMN tropism towards the inflammatory sites
have been the subject of extensive investigation in recent years
(Springer, 1994). In the initial phase, the rolling of leukocytes on
the endothelium is a prerequisite for subsequent adhesion (Von Andrian
et al., 1992; Lawrence and Springer, 1991) and it is mainly mediated by the selectins (Lasky, 1992; Abbassi et al.,
1993); interaction between the 
2-integrins on
the leukocyte membrane and their counterpart (members of the
immunoglobulin superfamily) on the endothelium sustains the firm
adhesion (Hynes, 1992; Carlos and Harlan, 1990). Less is known about
the actual emigration process, as characterized by pseudopodia emission
and opening of the endothelial gaps, although it is clear that it
requires G-protein-mediated cell activation (Springer, 1994).
Among the selectins, L-selectin (CD62L) is constitutively expressed on
PMN membrane and is required for the rolling of cells, mediated by
carbohydrate ligands, on the endothelial surface. P-selectin (CD62P)
and E-selectin (CD62E) are expressed on the endothelium under different
molecular mechanisms: P-selectin expression is both constitutive and
inducible upon treatment with several agents, e.g.,
histamine (Asako et al., 1994); E-selectin is expressed, at
least in vitro, only after stimulation of the endothelium
with pro-inflammatory cytokines (Springer, 1994).
We have recently characterized the existence of an endogenous pathway
that controls the PMN extravasation process and it is centered on the
action of leukocyte-derived lipocortin 1. Activation of this pathway by
exogenous administration of lipocortin 1 mimetics, such as its
N-terminus peptide acetyl 2-26 produces a significant inhibition of
PMN accumulation in distinct models of acute inflammation (Perretti
et al., 1993; Getting et al., 1997).
We describe a novel experimental mouse peritonitis model, in which a strong PMN accumulation is observed after challenge with MSU crystals. This was used to characterize the mechanisms of MSU crystal-induced PMN recruitment, mainly in the light of recent understandings of the mechanisms responsible for the initiation of the leukocyte extravasation process, i.e., role of endogenous mast cells and of endothelial-derived selectins. The effect of the lipocortin 1-derived peptide Ac2-26 was also assessed to investigate the potential therapeutic application of agents that activate the lipocortin 1 pathway in gout.
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Materials and Methods |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Animals. Male Swiss Albino mice (20-22 g body weight) were purchased from Banton and Kingsman (T.O. strain; Hull, Humberside, UK), and maintained on a standard food pellet diet with tap water ad libitum using a 12:00 hr light/dark cycle. Animals were used 3 to 4 days after the arrival.
MSU-induced PMN recruitment. The peritonitis was induced by injection of 1 to 5 mg MSU crystals, in 0.5 ml PBS (0.1 M, pH 7.4). At different time-points, animals were euthanized by CO2 exposure, peritoneal cavities washed with 3 ml of PBS containing 3 mM EDTA and 25 U/ml of heparin. Aliquots of the lavage fluids were then stained with Turk's solution (0.01% crystal violet in 3% acetic acid) and differential counting performed using a Neubauer hemacytometer and a light microscope (Olympus B061). Mononuclear cells and PMN could be easily identified. The large predominance of neutrophils in the PMN population in 6-hr lavage fluids was confirmed in cytospin preparations stained with May-Grunwald and Giemsa, confirming that >95% of PMN were neutrophils (not shown). Data are reported as 106 PMN/mouse.
Lavage fluids were then centrifuged at 400 × g 10 min and supernatants stored at -20°C before evaluation of the-glucuronidase activity (see below). Protein contents of cell-free
lavage fluids was also determined after staining in Blue Coomassie.
Readings were made at 570 nm wavelength in a plate-reader, and
absorbance values were converted to mg/ml after comparison with a
standard curve constructed with bovine serum albumin (0-5 mg/ml).
Other model of peritonitis. CPPD crystal- or zymosan-induced PMN recruitment were assessed either at 6 or 24 hr postchallenge. The dose of 3 mg was chosen for each of these insoluble phlogogens since comparison with an identical dose of MSU crystals was made. The number of PMN recruited into the murine peritoneal cavities was quantified as described above.
Endogenous mast cell.
The role played by resident peritoneal
mast cells on PMN accumulation elicited by MSU crystal injection was
validated by depleting animals of their endogenous mast cells, a
procedure already employed successfully to delineate the role of the
chemokine interleukin-8 (Perretti et al., 1994). As adapted
from Diaz et al. (1996), compound 48/80 was given i.p. (10 µg in 0.5 ml) 72 hr before MSU crystals, a time chosen to assure that
any acute inflammatory response elicited by the mast cell activator
would have subsided. Control mice received sterile PBS. PMN recruitment
was assessed 6 hr after MSU crystal (3 mg) injection as described
above.
). Aliquots (0.5 ml) of cell suspension were
taken at 30, 60 and 120 min, centrifuged at 1000 r.p.m. × 10 min
in a minifuge: supernatants were discarded and the resulting pellets
were stained overnight with 100 µl Toluidine blue solution. The
number of intact mast cells was then determined using a Neubauer
hemacytometer and light microscopy.
Drug treatment.
The H1-antagonists,
tripolidine (0.5 mg/kg) and diphenhydramine (9 mg/kg), or the PAF
receptor antagonist WEB2086 (10 mg/kg) were given i.p. simultaneously
with the local injection of MSU crystals (3 mg) (Harris et
al., 1996). Peptide Ac2-26 was given s.c. and the dose (200 µg
per mouse) was chosen from a recent study in which the peptide
effectively inhibited PMN accumulation into the mouse peritoneal cavity
in response to challenge with zymosan (Getting et al.,
1997). The mAbs were administered i.v. according to published protocols
(Rosen and Gordon, 1987; Watson et al., 1991). Mice were
treated i.v. with 100 µg rat anti-mouse CD11b mAb, rat anti-mouse
P-selectin or rat anti-mouse E-selectin mAb 1 hr before challenge with
MSU crystals (3 mg). Control animals received an equal dose of
nonimmune rat IgG. The sulfated polysaccharide, fucoidin, was given
i.v. 2 hr before MSU crystals at the dose of 0.3 mg per mouse (Harris
et al., 1995). In most cases PMN accumulation was evaluated
at the 6 hr time-point. However, the different role of selectins was
investigated also at 24 hr post-MSU crystal challenge: in this case,
mAbs were given i.v. 6 hr after the crystals, and PMN accumulation was
quantified at the 24 time-point as described above.
-Glucuronidase activity.
-Glucuronidase activity in
the supernatants was measured according to a published protocol
(Iwamura et al., 1993). A total of 250 µl of cell-free
lavage fluids was incubated with the substrate phenolphthalein--glucuronic acid (1 mM) in 0.5 ml total volume and
kept in a water bath at 37°C with gentle shaking for 18 hr. Reactions
were stopped by addition of 1 ml of ice cold glycine buffer (200 mM) in
200 mM NaCl (pH 10.4). Absorbance values, measured at 550 nm using a
96-well plate multi-reader, were transformed into U/ml of lavage fluid,
using a standard curve constructed with 0 to 2000 U -glucuronidase.
Data are reported as U per × 106 cells per
mouse.
Materials.
MSU and CPPD crystals were prepared by a
previously described method (Roberge et al., 1994; Denko and
Whitehouse, 1976). A boiling solution of 0.03 M MSU, pH 7.5, was
prepared by dissolving equimolar quantities of uric acid and sodium
hydroxide and filtering with an Acropor membrane filter (AN-3000, 3 µM; Gelman, Ann Arbor, MI). Sodium chloride (0.1 M final
concentration) was added to accelerate and improve the uniformity of
the crystallization. CPPD was obtained by mixing a calcium nitrate
solution (0.1 M final concentration) with an acidic solution of sodium
pyrophosphate (0.025 final concentration in
Na2P2O7
and 0.03 M HNO3). The milky white precipitate
formed CPPD crystals after 1 day at 50 to 60°C. The crystals were
characterized by x-ray diffraction (Rigaku Geirflex D/max), by
examination under phase and polarizing microscopy and by scanning
electron microscopy. The MSU and CPPD crystals showed triclinic
morphological characteristics. Their dimensions, determined by scanning
microscopy, were 10 × 1 × 1 µm to 25 × 1.5 × 1.5 µm, and 12 × 1.4 × 1.4 to 25 × 1.7 × 1.7 µm for MSU and CPPD, respectively. Both preparations were free of
endotoxin as determined by the Limulus Assay (Whittaker, Walkersville,
MS). The in vitro efficacy of these crystals on human PMN
has been recently validated (Hachica et al., 1995).
-glucuronic acid, glycine buffer, -glucuronidase
and all other chemicals were obtained from Sigma Chemical Co.
(Poole, U.K.). mAbs, rat anti-murine P-selectin (clone 5H1) and
E-selectin (clone 9A9) were produced at Hoffman-La Roche as described
(Norton et al., 1993; Labow et al., 1994), and
contained less than 1 endotoxin U/mg. An isotype-matched control rat
IgG was also prepared at Hoffman-La Roche. The rat anti-murine CD11b
mAb (clone C5.6) was purchased from Serotec (Oxford, UK).
Tripolidine-HCl and diphenhydramine-HCl were purchased from Research
Biomedical International (Natick, MA), whereas WEB2086 was a generous
gift of Boheringer Ingleheim (Ingleheim am Rheim, Germany).
Statistics.
Statistical differences were calculated on
original values using analysis of variance followed by Bonferroni test
for intergroup comparisons (Berry and Lindgren, 1990), or by unpaired
Student's t test (two-tailed) when only two groups were
compared. A threshold value of P < .05 was taken as significant.
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Results |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Characterization of MSU crystal-induced peritonitis.
Intraperitoneal injection of MSU crystals produced an intense PMN
accumulation at the 6 hr time-point (fig.
1A). The dose-response curve was
relatively steep, with a maximal effect (ranging between 8 and 12 × 106 PMN per mouse, n = 32) at
the 3-mg dose and a lower effect seen at the highest dose tested of 5 mg (probably due to a toxic action of the crystals). The dose of 3 mg
was selected for the subsequent experiments.
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-glucuronidase release after MSU
crystal injection. In the case of both parameters, maximal values were
obtained at 2 hr post-MSU challenge with subsequent lower amounts being
detected at the other time-points.
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Role of endogenous mast cells in MSU crystal-induced
inflammation.
A number ranging from 10 to 13 × 103 of intact mast cells were routinely recovered
from untreated mice (n = 10). Local administration of
compound 48/80 (10 µg) produced a marked depletion in intact mast
cells as assessed 72 hr postinjection: only 0.8 × 103 cells could be counted (n = 6; P < .05). The effect of this depletion treatment on MSU
crystal-induced leukocyte accumulation was then evaluated. A remarkable
reduction (58%) in the 6-hr PMN influx was observed when MSU crystals
were injected into mice pretreated with compound 48/80 compared to PBS
pretreated animals (fig. 2A). This
indicates that resident mast cells are playing an important role in the
cellular response activated by these crystals. To substantiate this
proposition, the effect of MSU crystals on the number of intact mast
cells recovered from the peritoneal cavities was quantified. Figure 2B
shows that this cell type is relatively sensitive to manipulation such
that a reduction in number was also seen after i.p. injection of 250 µl PBS. However, 3 mg of MSU crystals substantially diminished
(>95% reduction) the number of intact mast cells found at 2 hr
postinjection. As expected, values remained low for the entire period
under observation (up to 24 hr) (fig. 2B).
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Role of endogenous histamine and PAF in MSU crystal-induced
inflammation.
Treatment of mice with the PAF antagonist, WEB2086,
reduced significantly the 6 hr PMN accumulation observed in response to MSU crystal challenge by more than 50% (fig.
3). A similar effect was achieved by
pretreating mice with the H1 antagonist,
tripolidine (47% reduction, n = 17, P < .05).
Interestingly, a combined treatment with both antagonists did not
produce an additive effect, such that a 67% of inhibition of PMN
influx was then measured (n = 6, not significant
vs. either single treatment) (fig. 3). The involvement of
H1 receptors in the MSU response was validated further using diphenhydramine: 10.3 ± 0.20 × 106 PMN were recovered 6 hr after MSU crystal
challenge in control mice, and this figure was reduced to 4.70 ± 0.31 × 106 cells in animals pretreated with
diphenhydramine (9 mg/kg i.p.) (54% reduction, n = 6;
P < .05).
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Role of endothelial selectins in MSU crystal-induced inflammation. Administration of fucoidin (0.3 mg per mouse i.v., corresponding to ~10 mg/kg, -2 hr) significantly reduced MSU crystal-induced PMN accumulation (mean ± S.E.M.): 8.6 ± 0.2 × 106 PMN per mouse in saline-pretreated mice (n = 6) and 3.3 ± 0.3 × 106 PMN in fucoidin-pretreated animals (62% reduction; n = 10; P < .05).
The role of endothelial selectins was then investigated. Intravenous injection of anti-CD62P mAb alone significantly attenuated by 35% the 6-hr cell influx (n = 12, P < .05) (fig. 4A). A similar degree of inhibition was also seen in the group of mice treated with the anti-CD62E mAb (45% reduction, n = 10; P < .05), whereas almost a 70% reduction in PMN accumulation was measured after a combined treatment with both mAbs (n = 13; P < .05 vs. control IgG, and P < .05 vs. anti-CD62P mAb alone) (fig. 4A).
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Effect of anti-CD11b mAb and peptide Ac2-26 on MSU crystal-induced
inflammation.
A potent inhibition of PMN accumulation (-60%,
n = 6; P < .05) was seen after s.c.
administration of the lipocortin 1 pharmacophore, peptide Ac2-26 (fig.
5). A similar degree of inhibition was
also measured after treatment of mice with the anti-CD11b mAb (-68%, n = 10; P < .05) .
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Discussion |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Gouty arthritis is a neutrophil driven disease triggered by
deposition of MSU crystals in the joint space (McCarthy et
al., 1962; Brandt and Schumaker, 1995; Terkeltaub, 1992). The
ability to reproduce experimentally the marked cellular influx in
response to MSU crystals represents a convenient way to investigate the molecular mechanisms underlying this specific pathology. Previous studies have reported that the rat pleural cavity (Sedgwick et al., 1985) or a preformed rat air-pouch (Brooks et al.,
1987) provide convenient tissue sites to observe PMN influx in response to challenge with insoluble crystals. Here, we report that an i.p.
injection of MSU crystals causes a remarkable peritoneal accumulation
of murine PMN. Importantly, the cellular infiltration into the cavity
was sustained up to 24 hr, and this is different from other models of
acute inflammation in which insoluble particles are injected
(i.e., zymosan peritonitis). Another validation of this
experimental system came from the inability of CPPD crystals to produce
a rapid PMN influx similar to that obtained after challenge with MSU.
All these characteristics prompted us to begin an investigation into
the molecular mechanisms responsible for the intense and sustained
accumulation of PMN elicited by MSU crystals in our murine model.
PMN accumulation in response to MSU crystal injection was clearly
dissociated from plasma protein extravasation and -glucuronidase release, both peaking at 2 hr post-challenge. It therefore means that
the enzymatic activity is mainly derived from the resident macrophage
(Adams and Hamilton, 1992), rather than from the infiltrating PMN
(Iwamura et al., 1993).
It is well known that resident mast cells are in close proximity to the
site of leukocyte extravasation (i.e., the postcapillary venule) (Granger and Kubes, 1994), and recent studies have shown that
this cell type plays a central role in the initiation of the PMN
recruitment process (reviewed in Kubes and Granger, 1996). Mast cells
may release an array of mediators (histamine, PAF, pluripotent
cytokines, etc.) which are likely to activate the endothelium to
attract the circulating leukocyte, cause arrest and direct it to the
site of inflammation. We showed that a drastic reduction of resident
intact mast cell numbers in the mouse peritoneal cavity, achieved by
administration of compound 48/80 according to a well-established
protocol (Diaz et al., 1996), produced a marked suppression
of PMN accumulation in response to MSU crystal challenge. The ability
of these crystals to activate mouse mast cells was confirmed by the
dramatic suppression (>90%) of intact mast cell counts following
in vivo administration and, mainly, by the information
obtained with in vitro experiments. With the latter protocol
we could detect that MSU crystals produced a maximal mast cell
degranulation within 30 min of addition.
We then sought to identify, at least in part, which mast cell mediators
could contribute to the PMN influx generated by crystal injection. Mast
cell-derived histamine has been shown to induce leukocyte rolling on
the endothelium of postcapillary venules via up-regulation of
P-selectin on the endothelial cell surface (Asako et al.,
1994). The same study showed that this was effected by activation of
H1 receptors. In our study, when mice were
pretreated with the potent and specific H1
antagonist tripolidine, a remarkable attenuation of MSU-induced PMN
accumulation into the peritoneal cavity was observed. Similar data have
also been obtained with a chemically unrelated H1
antagonist, diphenhydramine. These observations clearly point to mast
cell-derived histamine as to a mediator involved in the cellular
response to MSU crystals.
A more recent study has described mast cell-derived PAF as a mediator
able to induce leukocyte adhesion to the post-capillary endothelium
(Gaboury et al., 1996). PAF action is mediated by 2-integrins, because an anti-CD18 mAb
suppresses PAF-induced cell adhesion (Kubes et al., 1990).
The effect of PAF antagonist, WEB2086 (Casals-Stenzel et
al., 1987), was tested in this study, finding again a significant
inhibition of PMN recruitment activated by MSU crystals. Importantly, a
combined treatment with tripolidine and WEB2086 did not produce an
additive effect. Because the rolling phenomenon is a prerequisite to
subsequent firm adhesion (Lawrence and Springer, 1991), these data
confirm the suggestion that, in the inflammatory response activated by
MSU crystals, mast cell-derived histamine and PAF are mediating
leukocyte rolling and adhesion, respectively, in a sequential manner.
The ability of fucoidin to inhibit PMN accumulation caused by MSU
crystals indicated that endogenous selectins were involved in the
process of cell recruitment (Lindbom et al., 1992). In murine systems a redundancy of endothelial-derived selectins has been
found, such that both adhesion molecules need to be blocked, with
specific mAbs, to inhibit PMN extravasation in experimental inflammation (reviewed in Ley and Tedder, 1995). P-selectin (CD62P), but not E-selectin (CD62E), blockade alone is sufficient per se only in
the initial phases of the inflammatory response activated by
thioglycollate (4 hr time-point) (Labow et al., 1994) or by the chemokine KC (2 hr time-point) (Harris et al., 1996),
when PMN recruitment is really modest. It was therefore somehow
surprising to observe that the intense PMN accumulation seen 6 hr
post-MSU crystal challenge could be inhibited either with the
anti-CD62P mAb or with the anti-CD62E mAb. This indicates not only that
both selectins are expressed after challenge with these crystals, but also that they are acting through different pathways (or by interacting with distinct ligands). Indeed, a combined treatment with both mAbs
produced essentially an additive inhibitory effect. A different scenario was seen when PMN recruitment at times later than 6 hr was
investigated. In this case the phenomenon of redundancy was observed
and only a combined treatment with both mAbs was able to suppress PMN
influx into the murine peritoneal cavities (probably they are now
interacting with an identical ligand). It is of interest that the
degree of inhibition attained (60%) was similar to that observed with
the combined treatment at the 6 hr time-point (~70%). The fact that
high PMN counts are found at 24 hr post-MSU crystal challenge, and that
this is inhibited with antiselectin mAbs, suggests that active cell
recruitment is occurring in the 6- to 24-hr time period. These data are
in agreement with a recent study conducted in the pig skin, in which
CD62E induction within the 4- to 24-hr time interval after intradermal
administration of MSU crystals was demonstrated (Chapman et
al., 1996).
We have previously compared the ability of an anti-CD11b mAb to inhibit
PMN recruitment in murine models of experimental inflammation with that
of the lipocortin 1 pharmacophore, the N-terminus derived peptide
Ac2-26 (Perretti et al., 1993). Both agents were also tested in our study, observing again a similar degree of inhibition (60-70%). The result of the anti-CD11b mAb experiment adds MSU crystals to the list of inflammatory stimuli that cause mouse PMN
recruitment via the 2-integrin
CD11b/CD18. This list includes zymosan, C5a, interleukin-1 and CXC
chemokines specific for PMN (Perretti et al., 1993; Harris
et al., 1995). Similarly, the efficacy of the lipocortin 1 mimetic peptide Ac2-26 may suggest, at least theoretically, a
potential therapeutic application for drugs arising from this line of
research. Gout is a disease characterized by massive neutrophil
accumulation into the joint space, and the data generated in this study
may suggest that this pathology could be susceptible to systemic
administration with lipocortin 1 mimetics. Modulation of endogenous
lipocortin 1 levels in circulating leukocytes by antiinflammatory
glucocorticoid hormones (Perretti and Flower, 1996) may account, at
least in part, for their therapeutic application in acute gout (Brandt
and Schumaker, 1995).
In conclusion, we describe a novel murine model of MSU crystal-induced inflammation. Taking advantage of 1) the simplicity of the model, 2) the intense and sustained PMN accumulation observed and 3) the availability of specific tools, we have attempted to shed light on mechanism(s) which may be operating in the pathology of gout.
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Footnotes |
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Accepted for publication June 2, 1997.
Received for publication February 24, 1997.
1 This work was supported by an endowment to the William Harvey Research Institute by the Ono Pharmaceutical Co. (Osaka, Japan). R.J.F. is a Principal Research Fellow of the Wellcome Trust. L.P. is supported by grants from Consiglio Nazionale delle Ricerche nos. 95.02383.CT04 and 96.03339.CT04. M.A.M. is recipient of a CNPq fellowship (Brasil).
Send reprint requests to: Dr. Mauro Perretti, Department of Biochemical Pharmacology, The William Harvey Research Institute, Charterhouse Square, London EC1 M 6BQ, United Kingdom.
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Abbreviations |
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CPPD, calcium pyrophosphate dihydrate; EDTA, ethylenediaminetetraacetic sodium salt; mAb, monoclonal antibody; MSU, monosodium urate; peptide Ac2-26, lipocortin 1-derived N-terminus peptide; PBS, phosphate-buffered saline; PAF, platelet-activating factor; PMN, polymorphonuclear leukocyte.
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References |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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2-integrin-mediated neutrophil adhesion at physiological shear rates in vivo.
Am. J. Physiol.
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 C. W. Lam, S. J. Getting, and M. Perretti In Vitro and In Vivo Induction of Heme Oxygenase 1 in Mouse Macrophages following Melanocortin Receptor Activation J. Immunol., February 15, 2005; 174(4): 2297 - 2304. [Abstract] [Full Text] [PDF]
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 S. J. Getting, C. Di Filippo, C. W. Lam, F. Rossi, and M. D'Amico Investigation into the Potential Anti-Inflammatory Effects of Endothelin Antagonists in a Murine Model of Experimental Monosodium Urate Peritonitis J. Pharmacol. Exp. Ther., July 1, 2004; 310(1): 90 - 97. [Abstract] [Full Text] [PDF]
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S. J. Getting, H. B. Schioth, and M. Perretti Dissection of the Anti-Inflammatory Effect of the Core and C-Terminal (KPV) {alpha}-Melanocyte-Stimulating Hormone Peptides J. Pharmacol. Exp. Ther., August 1, 2003; 306(2): 631 - 637. [Abstract] [Full Text] [PDF]
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S. J. Getting, H. C. Christian, C. W. Lam, F. N. E. Gavins, R. J. Flower, H. B. Schioth, and M. Perretti Redundancy of a Functional Melanocortin 1 Receptor in the Anti-inflammatory Actions of Melanocortin Peptides: Studies in the Recessive Yellow (e/e) Mouse Suggest an Important Role for Melanocortin 3 Receptor J. Immunol., March 15, 2003; 170(6): 3323 - 3330. [Abstract] [Full Text] [PDF]
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 S. J. Getting, G. H. Allcock, R. Flower, and M. Perretti Natural and synthetic agonists of the melanocortin receptor type 3 possess anti-inflammatory properties J. Leukoc. Biol., January 1, 2001; 69(1): 98 - 104. [Abstract] [Full Text]
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 P. G. McLean, A. Ahluwalia, and M. Perretti Association between Kinin B1 Receptor Expression and Leukocyte Trafficking across Mouse Mesenteric Postcapillary Venules J. Exp. Med., July 31, 2000; 192(3): 367 - 380. [Abstract] [Full Text] [PDF]
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S. J. Getting, L. Gibbs, A. J. L. Clark, R. J Flower, and M. Perretti POMC Gene-Derived Peptides Activate Melanocortin Type 3 Receptor on Murine Macrophages, Suppress Cytokine Release, and Inhibit Neutrophil Migration in Acute Experimental Inflammation J. Immunol., June 15, 1999; 162(12): 7446 - 7453. [Abstract] [Full Text] [PDF]
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