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4 Integrin to Recruit Leukocytes to the Central Nervous System in Experimental Autoimmune Encephalomyelitis1
Immunology Research Group, Department of Physiology and Biophysics, University of Calgary, Calgary, Alberta, Canada
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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4 integrin increased with time
as anti-4 integrin blocked 
20, 50, and 60% of
leukocyte rolling 2 days before disease onset, 5 days and 2 wk
postonset of symptoms, respectively, and 85% of rolling 5 wk
postsymptoms. Addition of anti-P-selectin to 4
integrin Ab-treated mice blocked all remaining rolling at each time
point. Interestingly, however, 4 integrin-mediated
rolling appeared to be entirely dependent on P-selectin as
anti-P-selectin alone was able to completely block all leukocyte
rolling. In the absence of rolling (with P-selectin Ab), a 70%
reduction in adhesion was noted. A very similar reduction was seen when
mice were treated with 4 integrin-blocking Ab. In
conclusion, we describe increased leukocyte trafficking in the brains
of EAE mice with important overlapping roles for both P-selectin and
4 integrin in mediating leukocyte-endothelial cell
interactions. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The mechanisms of leukocyte recruitment to the CNS are not well
understood. The current paradigm of recruitment has largely been worked
out in vitro and in a few easily accessible tissues in vivo
(mesentery, cremaster, and skin) in which intravital microscopy can be
performed. Circulating leukocytes first tether to and then roll
on endothelial cells expressing adhesion molecules. If appropriate
signals are encountered, leukocytes then firmly adhere to the vessel
wall before transmigrating into the surrounding tissue
(1, 2, 3). The tethering and rolling steps are mediated by
the selectins, as well as 4 integrins. Firm
adhesion occurs when integrins are induced to bind their endothelial
ligands with high affinity. Until very recently, in vivo investigations
of EAE have had to rely on histological observations and clinical
outcome following blockade of various adhesion molecules to infer the
mechanisms of leukocyte recruitment to the CNS. These studies have
primarily focused on a role of 4 integrin.
Indeed, treatment with anti-4 integrin Abs
has some protective effects in EAE and can partially reduce the numbers
of inflammatory cells in the CNS of diseased animals
(4, 5, 6). However, because 4
integrin has additional roles to leukocyte recruitment, including T
cell and effector cell activation, the mechanism of action of
4 integrin blockade in EAE is not yet
clear.
A role for selectins in recruitment to the CNS has been discounted, largely due to their limited expression in the CNS microvasculature (7, 8). Despite this, recent evidence suggests that selectins may yet have a role in leukocyte recruitment to the CNS. Intravital microscopy of TNF- or LPS-treated mice revealed that P-selectin blockade reduced leukocyte rolling in brain microvessels (9, 10). Indeed, a recent study by Carrithers et al. (11) implicated P-selectin in the initial recruitment of activated encephalitogenic T cells into the healthy CNS. However, a role for P-selectin in the inflammatory stage of EAE has not been assessed to date.
We (9) and others (10, 12) have recently
developed intravital microscopy of CNS microvessels. Using this
technique, we for the first time directly investigate the mechanisms of
leukocyte recruitment to the CNS in EAE. Early in disease, P-selectin
played a principal role in mediating leukocyte rolling in brain
microvessels, which corresponded with an induction of P-selectin
expression in the CNS. As disease progressed,
4 integrin gained importance as the molecule
to mediate leukocyte rolling. Nevertheless, 4
integrin was insufficient to support rolling in the absence of
P-selectin. Inhibition of either P-selectin or
4 integrin reduced subsequent leukocyte
adhesion in EAE, supporting their potential role in the pathogenesis of
this disease.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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RB40.34 (anti-P-selectin) and R1-2
(anti-4 integrin) were purchased from BD
PharMingen (Mississauga, Ontario, Canada) as were
FITC-conjugated anti-murine CD3
(clone 145-2C11) and Mac-1
(clone M1/70). RME-1 (anti-E-selectin) was generously supplied by
Dr. A. Issekutz (Dalhousie University, Halifax, Nova Scotia, Canada).
Myelin oligodendrocyte glycoprotein (MOG ) 35–55 peptide was initially
a gift from Dr. C. Bernard (La Trobe University, Melbourne, Australia),
and later generated in our own peptide synthesis laboratory (University
of Calgary, Calgary, Alberta, Canada). Female C57BL/6 mice were
purchased from Charles River Breeding Laboratories (Montreal, Quebec,
Canada).
Induction of EAE
EAE was induced in mice as previously described (13). Briefly, 9–11-wk-old C57BL/6 mice were immunized s.c. with 50 µg of a peptide generated from MOG35–55 in CFA (Sigma-Aldrich, Oakville, Ontario, Canada). Mice were immunized twice, 1 wk apart. Pertussis toxin (200 ng; List Biological Laboratories, Campbell, CA) was injected i.p. on the day of the first immunization and then again 2 days later. Disease was monitored daily and scored as follows: 0, no clinical signs; 1, tail paralysis; 2, tail paralysis and hind-limb weakness; 3, hind limb paralysis; 4, complete hind limb paralysis and front limb weakness.
Intravital microscopy
Intravital microscopy of the mouse cerebromicrovasculature was performed as previously described (9). Briefly, the tail vein was cannulated for the administration of additional anesthetic, fluorescent dyes, and other reagents. A craniotomy was performed using a high-speed drill (Fine Science Tools, North Vancouver, British Columbia, Canada) and the dura matter was removed to expose the underlying pial vasculature. Throughout the experiment, the mouse was maintained at 37°C and the exposed brain was kept moist with an artificial cerebrospinal fluid buffer.
To observe leukocyte/endothelial interactions, leukocytes were fluorescently labeled by i.v. administration of rhodamine 6G (0.3 mg/kg body weight) and observed using a microscope (Axioskop, x10 eyepiece and x25 objective lens; Zeiss, Don Mills, Ontario, Canada) outfitted with a fluorescent light source (epi-illumination at 510–560 nm using a 590-nm emission filter). A low light intensifier charge-coupled device camera (Stanford Photonics, Palo Alto, CA) mounted on the microscope was used to project the image to a monitor. Three different postcapillary venules with a diameter between 30 and 70 µm were chosen for observation. All experiments were recorded for later analysis. Rolling leukocytes were defined as those cells moving at a velocity less than that of erythrocytes within a given vessel. Leukocytes were considered adherent if they remained stationary for 30 s or longer.
Experimental protocol
Leukocyte rolling and adhesion was observed by intravital microscopy at various time points corresponding to different phases of disease: 2–3 days before predicted onset of symptoms (10 days post first immunization with MOG35–55) corresponding to developing presymptomatic EAE, 4 days postdevelopment of symptoms corresponding to acute disease, and 2 and 5 wk postsymptoms corresponding to chronic disease. Depending on the experiment, 70 µg R1-2, 100 µg RME-1, or 20 µg RB40.34 in 200 µl saline were administered i.v. based on previous work from our laboratory showing that these are optimal concentrations of Ab needed to inhibit rolling in muscle microvessels. For experiments requiring pretreatment with adhesion molecule blocking Ab, Abs were administered at the same concentrations as described above. Five-hour pretreatment was used for R1-2 and 24-h pretreatment was used for RB40.34.
Flow-assisted cytometry (FACs) analysis of inflammatory cells in the CNS
Infiltrating inflammatory cells in the CNS were analyzed by FACs as previously described (14). Mice were perfused through the heart with PBS to clear circulating blood from the vasculature. Brain and spinal cord tissue of EAE mice were dissociated through a wire mesh. Mononuclear cells were then separated on a Percoll (Amersham Pharmacia Biotech, Baie d’Urfé, Quebec, Canada) gradient. Cells were then incubated with FITC-conjugated anti-CD3 or Mac-1 and analyzed with a BD Biosciences FACScan (Mountain View, CA). Untreated mice were used as controls to ensure that mononuclear cell counts were not simply due to vascular leukocyte content.
Dual-radiolabeled Ab assay for P-selectin expression
P-selectin expression was quantitatively measured in the brain and other tissues of healthy control and EAE mice by the dual-radiolabeled Ab assay as previously described (9, 15). Briefly, a mixture of 10 µg 125I-labeled anti-P-selectin (RB40.34) and a dose of 131I-labeled nonbinding Ab (A110-1, anti-keyhole limpet hemocyanin) calculated to achieve a total 131I activity of 4–6 x 105 cpm were injected i.v. through the jugular vein. The Abs were allowed to circulate for 5 min. A blood sample was taken from the carotid artery and the mouse was then completely perfused with saline through the jugular vein to remove all circulating blood. Tissues were harvested, weighed, and measured for 125I and 131I activity. P-selectin expression was calculated per gram of tissue by subtracting accumulated 131I activity (labeled nonbinding Ab) from accumulated 125I activity (labeled anti-P-selectin). The 131I activity of the nonbinding Ab accounts for circulating Ab or protein that has leaked into tissue nonspecifically. Data are represented as the percentage of the injected dose of Ab per gram of tissue. This approach is sufficiently sensitive to detect small but significant amounts of constitutive P-selectin in wild-type relative to no P-selectin in P-selectin-deficient mice (15).
Statistics
Data in graphs is shown as mean ± SEM unless indicated otherwise. A Student’s t test with Bonferroni correction was used for multiple comparisons. Statistical significance was set at p < 0.05.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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EAE was induced in C57BL/6 mice as described in Materials and Methods. This protocol resulted in a chronic, nonremitting disease consistent with previous reports (16). Symptoms arose between 11 and 15 days following the first immunization with MOG35–55, and escalated rapidly over 6–7 days at which point they stabilized. Only very slight recovery was observed over the next 4 wk (data not shown).
Leukocyte rolling and adhesion is increased in the CNS microvasculature of EAE mice
We used intravital microscopy to directly observe
leukocyte-endothelial interactions in the cerebromicrovasculature of
EAE mice (Fig. 1
). Very little leukocyte
recruitment was observed in the brains of healthy control mice. Rolling
cells were almost never observed and adherent leukocytes were also rare
(Fig. 2). In contrast, profound
recruitment was observed in EAE mice. Approximately 30 cells/min were
observed rolling in postcapillary venules of presymptomatic mice (Fig. 2A). Rolling remained elevated over the next week,
significantly falling to <15 cells/min 2 wk postdevelopment of
symptoms and returning to near-normal levels 5 wk postdevelopment of
symptoms. Leukocyte adhesion was also significantly induced in
presymptomatic mice to 10-fold above control (Fig. 2B).
Adhesion peaked in the acute phase of disease and returned to near
normal levels 2 wk following symptom onset.
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Infiltrating inflammatory cells in the CNS of EAE mice were
determined by FACs (Fig. 3). Very few
inflammatory cells could be detected in healthy control mice. In
contrast, large numbers of lymphocytes and macrophages could be
detected in the CNS of mice with EAE. Inflammatory cells were detected
in greatest numbers in mice 4 days postdevelopment of symptoms, but
were also present in presymptomatic mice and mice 2 wk postdevelopment
of symptoms. The ratio between lymphocyte and macrophages (1:3) did
not change over the course of disease. Neither did the ratio between
CD3+ and CD3- cells in the
lymphocyte population favoring CD3+ cells to a
small degree (58%).
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4 integrin together mediate leukocyte
rolling in the cerebromicrovasculature of EAE mice
To determine the mechanism by which leukocytes are recruited to
the brain in EAE mice, intravital microscopy was used to directly
observe the affects of adhesion molecule blockade on leukocyte
recruitment. After baseline rolling was measured in mice at various
stages of disease, anti-4 integrin was
administered i.v. and leukocyte rolling was observed 20 min later. We
have previously shown that 20 min is required for optimal inhibition of
rolling by R1-2 (17) and that isotype-matched Ab has no
effect on leukocyte recruitment (18). Fig. 4 summarizes that the role for
4 integrin-mediated rolling increased over the
course of disease. Although 4 integrin
blockade decreased rolling by 20% in presymptomatic mice (Fig. 4A), this increased to 50% in mice in the acute (Fig. 4B, 4 days postsymptom onset) and 60% in chronic (Fig. 4C, 2 wk postsymptom onset) phases of disease. The rolling
cells observed in mice 5 wk postsymptom onset were almost completely
(90%) blocked by anti-4 integrin (Fig. 4D). Following blockade of 4
integrin, all remaining rolling was further blocked by
anti-P-selectin at all stages of disease (Fig. 4). We have
previously shown that an isotype-matched Ab has no effect on leukocyte
rolling (18). In a separate series of experiments, we
administered anti-P-selectin alone and surprisingly found that it
was sufficient to completely block all leukocyte rolling (Fig. 5A). This demonstrates that
4 integrin-mediated rolling is completely
dependent on P-selectin. This observation has been made before, that
4 integrin can support rolling but is unable
to tether the cells, an event requiring selectins (17).
Anti-E-selectin had no effect on leukocyte rolling (Fig. 5B), demonstrating that the blockade was specific for
P-selectin, not E-selectin.
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4 integrin mediates leukocyte adhesion in the brain
microvasculature
To determine the mechanisms mediating leukocyte adhesion in the
cerebromicrovasculature of EAE mice, mice were pretreated with
anti-4 integrin 5 h before adhesion
was observed by intravital microscopy. As
anti-4 integrin was unable to displace
already adherent cells (data not shown) pretreatment with the Ab was
required to prevent the accumulation of new cells. Mice in the acute
phase of disease (4 days postonset of symptoms) were investigated as
maximal adhesion was observed at this time point (Fig. 2
B).
Blockade of 4 integrin resulted in a 70%
reduction in leukocyte adhesion (Fig. 6B), demonstrating that
4 integrin is an important mediator of
adhesion in EAE. A similar reduction in adhesion was observed with 24-h
pretreatment with 4 integrin Ab (data not
shown). To determine whether leukocyte adhesion in the
cerebromicrovasculature was dependent on leukocyte rolling, acute-phase
mice were pretreated for 24 h with anti-P-selectin. Rolling
was completely blocked in these mice over the 24 h (Fig. 6A). Leukocyte adhesion was reduced by 70% (Fig. 6B).
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P-selectin protein expression in the brain and spinal
cord of EAE mice was investigated using a quantitative radiolabeled Ab
assay. Although completely absent in the CNS of healthy control mice, a
dramatic induction of P-selectin expression was observed in the brains
of EAE mice, with greatest expression in presymptomatic mice (Fig. 7A). Levels fell over the
course of disease, but remained significantly elevated compared with
control. P-selectin expression was variable in the spinal cords of
presymptomatic mice (Fig. 7B), and therefore did not reach
significance over healthy controls. However, similar to the brain,
significant expression was observed in mice 4 day and 2 wk
postdevelopment of symptoms.
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). Increased expression was
largely restricted to the CNS, with the exception of presymptomatic
mice which had significantly increased levels of P-selectin protein in
a number of abdominal organs including the liver, pancreas, spleen,
mesentery, and stomach. P-selectin was also up-regulated in the bone
marrow of mice 2 wk after development of symptoms. P-selectin
expression in the brain was very low compared with other tissues as
optimal expression in presymptomatic mice was lower than baseline
expression observed in tissues like the skin and muscle of healthy
control mice. Despite this, the relative increase in P-selectin
expression was greatest in the brain.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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4 integrin to rolling
increased dramatically and was dominant for adhesion. However, the role
for P-selectin has clearly been underappreciated as throughout disease
P-selectin was required for 4 integrin to
mediate rolling.
Initial work proposed that there was a limited role for selectins in
leukocyte recruitment to the CNS, due to limited selectin expression.
For example, Engelhardt et al. (7) reported a lack of
P-selectin and E-selectin expression in EAE. However, more recent work
has proposed a role for the selectins in the inflamed brain
microvasculature. Although Barkalow et al. (8) were unable
to see rapid P-selectin expression in cultured cerebral endothelial
cells, this group reported that these same cells did have the capacity
to synthesize P-selectin at a more delayed time point. Intravital
microscopy of the brain revealed that TNF induces P-selectin- and
E-selectin-dependent neutrophil recruitment in the brain
microvasculature (9). A very recent study by Piccio et al.
(10) suggested that TNF- or LPS-stimulated brain
microvasculature induced autoreactive T cells to roll and adhere. In
that study, the authors also used fluorescently labeled
anti-P-selectin Abs and demonstrated an increase in P-selectin
expression in brain vessels of EAE mice. Using an extremely sensitive
and quantitative approach to measuring P-selectin expression, we
demonstrated no basal P-selectin expression, but a very significant
increase in P-selectin expression before disease onset that remained
elevated for at least 2 wk postsymptoms. P-selectin was detected in
both the brain and the spinal cord, confirming that its expression was
not limited to the brain microvasculature. The potential explanation
for the lack of previous detection of cerebral P-selectin by some
groups may be related to the fact that our assay revealed that optimal
P-selectin expression in the brain was only at levels seen basally in
organs like muscle and skin (Table I). Nevertheless, these levels have
previously been reported to be sufficient to mediate basal rolling in
muscle and skin (19). Most importantly, our study
demonstrates a functional role for P-selectin in the leukocyte rolling
during EAE, particularly a few days before disease onset where
P-selectin mediated the large majority of rolling independent from
4 integrin. P-selectin maintained a role
throughout disease as with disease progression, rolling was mediated by
a combination of P-selectin and 4
integrin.
As the disease progressed, the importance of P-selectin as a rolling
molecule was replaced by the 4 integrin
pathway. This integrin is very unusual in its ability to induce
tethering, rolling, and adhesion thereby potentially bypassing the need
for the selectins. Indeed, Alon et al. (20) and Berlin et
al. (21) both demonstrated that immobilizing a ligand for
4 integrin within a laminar flow chamber was
sufficient to induce flowing lymphocytes to tether and roll, and when
an appropriate activation signal was provided, then the lymphocytes
were also able to firmly adhere. In vivo, 4
integrin was also shown to support rolling and adhesion in inflamed
mesenteric microvessels of adjuvant-treated rats (17), but
in this case selectins were also required. By contrast,
4 integrin did support tethering, rolling, and
adhesion in muscle microvessels treated with IL-4 in the absence of all
selectins (22). Clearly, the tissue and/or the
inflammatory process dictated whether 4
integrin required selectins. In our study, early in EAE development,
4 integrin appeared to have only a minor role
in the rolling process. By contrast, with progression of disease,
blockade of 4 integrin could reduce both
rolling as well as adhesion suggesting a more and more important role
for this molecule. However, 4
integrin-mediated rolling remained additionally dependent on
P-selectin, likely for initial leukocyte tethering.
The importance of 4 integrin is underscored by
the reduced symptoms in EAE mice associated with spinal cord disease
(4, 5, 6, 23). The fact that
anti-4 integrin blocks recruitment to the
brain may bode well for future therapeutic intervention in human
disease. However, blockade of 4 integrin is
not completely protective from disease, and we show in this study that
it cannot prevent all leukocyte adhesion in the brain microvasculature,
suggesting that other adhesion molecules may also have a role.
Interestingly, by blocking all leukocyte rolling with
anti-P-selectin Ab, it was also not possible to completely prevent
leukocyte adhesion. Rolling is usually considered to be a prerequisite
for adhesion, but recent studies have suggested that in some cases
leukocytes may be able to tether and immediately adhere, bypassing the
rolling stage. Vajkoczy et al. (12) recently showed that
activated T cells can adhere without rolling in spinal cord
microvessels via 4 integrin. This may be the
mechanism responsible for residual adhesion we observed in
anti-P-selectin-pretreated mice. Alternatively, other selectins
(E-selectin or L-selectin) may contribute. We observed no role for
E-selectin, but it is not clear whether L-selectin could contribute to
tethering. L-selectin-deficient mice are protected from EAE, however,
this may have more to do with the inability of
L-selectin-/- monocytes to migrate through the
tissue once across the vascular wall or even to recognize targets
rather than a defect in leukocyte recruitment per se (24).
Regardless, the inability of anti-P-selectin to block all leukocyte
adhesion in the CNS of EAE mice may explain why Engelhardt et al.
(7) did not observe any benefit in EAE mice. Based on our
work, we would propose dual inhibition of 4
integrin and P-selectin may provide optimal benefit in human disease.
In conclusion, our data demonstrate that during the development of EAE
in mice, early P-selectin-dependent leukocyte rolling is induced that
correlates with P-selectin expression just before symptom onset.
However, with the progression of disease, P-selectin expression is
somewhat reduced and the importance of the 4
integrin as a rolling molecule increases such that by 5 wk, this
molecule is able to support essentially all rolling and the majority of
adhesion. However, this 4 integrin-mediated
rolling remains entirely dependent on P-selectin, demonstrating the
importance of P-selectin to leukocyte recruitment to the CNS in EAE.
Finally, our data also suggest that even before disease onset when
P-selectin dominates as a key rolling molecule,
4 integrin mediates much of the observed
adhesion. Clearly, designing molecules to inhibit
4 integrin preferably with anti-selectins
may significantly impact upon the development of disease in EAE and
possibly in MS.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Paul Kubes, Immunology Research Group, Department of Physiology and Biophysics, University of Calgary, 3330 Hospital Drive NW, Calgary, Alberta, Canada T2N 4N1. E-mail address: pkubes{at}ucalgary.ca
3 Abbreviations used in this paper: MS, multiple sclerosis; EAE, experimental autoimmune encephalomyelitis; MOG, myelin oligodendrocyte glycoprotein; FACs, flow-assisted cytometry.
Received for publication February 20, 2002. Accepted for publication May 20, 2002.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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4
1 integrin. Nature 356:63.[Medline]
4-integrin but not 47-integrin. J. Clin. Invest. 102:2096.[Medline]
4 integrin suppresses and reverses active experimental allergic encephalomyelitis. J. Neuroimmunol. 58:1.[Medline]
-induced leukocyte recruitment in the brain microvasculature. Circ. Res. 87:1141.4-integrin-VCAM-1 binding mediates G protein-independent capture of encephalitogenic T cell blasts to CNS white matter microvessels. J. Clin. Invest 108:557.[Medline]
4-integrin supports leukocyte rolling and adhesion in chronically inflamed postcapillary venules in vivo. J. Exp. Med. 183:1995.4-integrin, P-selectin, and E-selectin in an allergic model of inflammation. J. Exp. Med. 185:1077.4 integrins mediate lymphocyte attachment and rolling under physiologic flow. Cell 80:413.[Medline]
4 integrin. J. Immunol. 163:3441.4, but not L-selectin, prevent central nervous system inflammation and experimental encephalomyelitis by blocking secondary leukocyte recruitment. Proc. Natl. Acad. Sci. USA 96:6896.This article has been cited by other articles:
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|
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|
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|
 |
|
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|
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|
|
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|
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|
|
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|
|
|
|
|
|
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|
|
|
|
 |
|
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|
|
|
|
 |
|
 W Qi, K V J Ebbert, A W B Craig, P A Greer, and D-M McCafferty Absence of Fer protein tyrosine kinase exacerbates endotoxin induced intestinal epithelial barrier dysfunction in vivo Gut, August 1, 2005; 54(8): 1091 - 1097. [Abstract] [Full Text] [PDF]
|
|
|
|
|
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|
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|
|
|
|
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|
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|
|
|
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 P. Kivisakk, D. J. Mahad, M. K. Callahan, C. Trebst, B. Tucky, T. Wei, L. Wu, E. S. Baekkevold, H. Lassmann, S. M. Staugaitis, et al. Human cerebrospinal fluid central memory CD4+ T cells: Evidence for trafficking through choroid plexus and meninges via P-selectin PNAS, July 8, 2003; 100(14): 8389 - 8394. [Abstract] [Full Text] [PDF]
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 L. Battistini, L. Piccio, B. Rossi, S. Bach, S. Galgani, C. Gasperini, L. Ottoboni, D. Ciabini, M. D. Caramia, G. Bernardi, et al. CD8+ T cells from patients with acute multiple sclerosis display selective increase of adhesiveness in brain venules: a critical role for P-selectin glycoprotein ligand-1 Blood, June 15, 2003; 101(12): 4775 - 4782. [Abstract] [Full Text] [PDF]
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 U. H. von Andrian and B. Engelhardt {alpha}4 Integrins as Therapeutic Targets in Autoimmune Disease N. Engl. J. Med., January 2, 2003; 348(1): 68 - 72. [Full Text] [PDF]
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W. G. James, D. C. Bullard, and M. J. Hickey Critical Role of the {alpha}4 Integrin/VCAM-1 Pathway in Cerebral Leukocyte Trafficking in Lupus-Prone MRL/faslpr Mice J. Immunol., January 1, 2003; 170(1): 520 - 527. [Abstract] [Full Text] [PDF]
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