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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
From the Department of Biomedical Engineering,
University of Virginia, Charlottesville, and the Howard Hughes Medical
Institute, Department of Cellular and Molecular Medicine, University of
California, La Jolla.
Leukocyte capture and rolling are mediated by selectins expressed
on leukocytes (L-selectin) and the vascular endothelium (P- and
E-selectin). To investigate the role of core 2 Leukocyte recruitment to sites of inflammation
requires a multistep adhesion cascade beginning with leukocyte capture
and rolling leading to firm adhesion and
transmigration.1,2 Capture and rolling are mediated
largely by the selectin family of adhesion molecules. Intravital
microscopy studies on inflamed tissue of the cremaster muscle conducted
in gene-targeted mice deficient in either E-, P-, or L-selectin or
various combinations have revealed both overlapping and unique
functions of the selectins.3,4 The overlapping function of
selectins is in particular evident in E-selectin-deficient mice. These
mice do not show obvious inflammatory deficits in certain models unless
P-selectin is also blocked,5 which then leads to a
dramatic reduction in the number of rolling leukocytes.6
Characteristic properties of individual selectins become evident in
their ability to mediate leukocyte rolling at distinct velocities.
Untreated wild-type mice show P-selectin-dependent rolling with
typical rolling velocities in venules of the cremaster muscle of below
50 µm/s.7 In tumor necrosis factor- Selectin ligands carry sialylated and fucosylated oligosaccharides
found at the nonreducing termini of core 2-dependent O-linked glycans.8-10 Core 2 structures are formed by a family of
core 2 The present study was designed to test whether P- and/or E-selectin
ligand functions are impaired under physiological conditions in vivo by
the absence of C2GlcNAcT-I. To this end, we investigated leukocyte
rolling in vivo in C2GlcNAcT-I-deficient mice in 2 models of
inflammation (untreated and TNF Animals
Antibodies and cytokines
Flow cytometry Flow cytometry was used to find the saturating dose of PSGL-1-blocking mAb 4RA10 after systemic injection into mice. Four anesthetized control mice were treated with increasing doses of mAb 4RA10 (1 µg, 3 µg, 10 µg, and 100 µg per mouse) or a rat antimouse IgG control antibody (100 µg) (Pharmingen, San Diego, CA). At 20 minutes after injection of the PSGL-1-blocking mAb 4RA10 or the isotype control, peripheral blood was collected from the carotid artery. Red blood cells were lysed in Pharm-Lyse-10X solution (Pharmingen). After centrifugation and removal of the supernatant, blood cells (1 × 106 cells) were suspended in phosphate-buffered saline (PBS)-1% bovine saline solution and incubated with a mouse antirat secondary antibody conjugated with flourescein isothiocyanate (FITC) (Pharmingen) for 30 minutes on ice (1 µg/106 cells). In addition, mAb Ly-6G against the neutrophilic surface antigen GR-1 conjugated with phycoerythrin (Pharmingen) was added to gate for neutrophilic granulocytes. PSGL-1 expression was determined on 10 000 leukocytes per mouse by means of a 4-decade FACScan with Cell Quest software package (Becton Dickinson, San Jose, CA).Western blotting Bone marrow cells were flushed from the femurs and tibias of control and core 2 / mice by means of cold RPMI 1640 (Gibco BRL, Grand Island, NY) and immediately centrifuged at 1000 rpm
for 5 minutes. The pellet was washed once with cold PBS and extracted
with lysis buffer (108 cells/mL) containing 1% Triton
X-100 (Fisher Scientific, Pittsburgh, PA) along with 1 mM phenylmethyl
sulfonyl fluoride, 1 mM egtazic acid, and 1% protease inhibitor
cocktail (all Sigma Chemical, St Louis, MO) in PBS, pH 7.4. Extraction
was carried out at 4°C for 30 minutes on an orbital shaker. The
samples were clarified by centrifugation at 14 000g for 30 minutes at 4°C, and the supernatant was transferred to fresh tubes.
Samples were boiled for 5 minutes in sodium dodecyl sulfate-polymerase
chain reaction sample buffer with  -mercaptoethanol, electrophoresed
on a 10% polyacrylamide gel, and transferred to nitrocellulose. After
blocking with 5% nonfat milk in PBS for 1 hour at room temperature,
the membrane was probed with PSGL-1 mAb 4RA10 or rat isotype control (5 µg/mL, BD Pharmingen) in 2.5% nonfat milk and 0.05% Tween-20
(Fisher Scientific) in PBS for 1 hour at room temperature, washed 3 times with PBS-Tween 20 (0.05%), and then incubated with goat antirat IgG conjugated to horseradish peroxidase (Pierce, Rockford, IL) for 1 hour at room temperature. After 3 washes in PBS-Tween 20 (0.05%) followed by 1 wash in PBS, the blots were developed by enzyme
chemiluminescence by means of the substrate kit (Pierce) and were
exposed to X-omat AR5 film (Eastman Kodak, Rochester, NY).
Intravital microscopy Mice were anesthetized with an intraperitoneal injection of ketamine (125 mg/kg body weight) (Ketalar) (Parke-Davis, Morris Plains, NJ); xylazine (12.5 mg/kg body weight) (Phoenix Scientific, St Joseph, MO); and atropin sulfate (0.025 mg/kg body weight) (Elkins-Sinn, Cherry Hill, NJ) and placed on a heating pad to maintain body temperature. Some mice were pretreated with an intrascrotal injection of recombinant murine TNF as described.6
Microscopic observations were made on an intravital microscope (Axioskop) (Zeiss, Thornwood, NY) with a saline immersion objective (SW 40/0.75 numerical aperture). The trachea was intubated, and one jugular vein was cannulated for administration of anesthetic throughout the intravital microscopic experiment. One carotid artery was cannulated for blood pressure monitoring, blood samples, and systemic mAb injections. Blood pressure was monitored intermittently during the experiment (model BPMT-2) (Stemtech, Menomonee Falls, WI). During the experiment, mice received 0.2 mL/h diluted pentobarbital in saline to maintain anesthesia and a neutral fluid balance. Cremaster muscle preparation The cremaster muscle was prepared for intravital microscopy as described.6 The epididymis and testis were gently pinned to the side, exposing the well-perfused cremaster microcirculation. The cremaster muscle was superfused with thermocontrolled (35°C) bicarbonate-buffered saline. To detect changes in systemic white blood cell count after injection of the various antibodies, systemic blood samples (10 µL) were taken after each mAb injection. Blood samples were diluted 1:10 with Kimura (11 mL of 5% [wt/wt] toluidine blue. 0.8 mL of 0.03% light green SF yellowish, 0.5 mL saturated saponin, and 5 mL of 0.07 M phosphate buffer, pH 6.4) (all from Sigma) and were analyzed for leukocyte concentration (expressed as number of leukocytes per microliter of whole blood). Differential leukocyte counts were taken from blood smears by means of the Hema 3 Kit (Biochemical Sciences, Swedesboro, NJ). To differentiate intravascular and interstitial leukocytes into neutrophils, eosinophils, and mononuclear cells, whole mounts of cremaster muscle were prepared as described elsewhere.25 Both rolling and firmly adhered leukocytes are visible as intravascular cells in whole-mount histological preparations.26Data analysis Microvessel diameters, lengths, and rolling leukocyte velocities were measured by means of a digital image processing system.27 Each rolling leukocyte passing a line perpendicular to the vessel axis was counted, and leukocyte rolling flux was expressed as leukocytes per minute. Rolling flux fraction was calculated as described7 by dividing leukocyte rolling flux by total leukocyte flux estimated as [WBC] × vb × × (d/2)2
where [WBC] is the systemic leukocyte count, vb is the
blood flow velocity, and d is the venular diameter. Leukocyte rolling
velocities were measured as averages over a 2-second time window.
Rolling velocities of 5 leukocytes were measured in each venule.
Centerline red blood cell velocity in the cremaster muscle preparation
was measured by means of a dual photodiode and a digital on-line
cross-correlation program (Circusoft Instrumentation, Hockessin, DE).
Centerline velocities were converted to mean blood flow velocities by
multiplying with an empirical factor of 0.625.28 Wall
shear rates ( w) were estimated as 2.12 (8 vb/d), where
vb is the mean blood flow velocity, d is the diameter of the vessel,
and 2.12 is a median empirical correction factor obtained from velocity
profiles measured in microvessels in vivo.29
To specifically address the shear rate dependence of leukocyte rolling through E-selectin, rolling flux fractions were stratified and grouped by wall shear rate and used to construct Figure 5. Statistics Statistical analysis was performed with the Sigma-Stat 2.0 software package (SPSS Science, Chicago, IL). Average vessel diameter, leukocyte rolling flux fractions, leukocyte rolling velocities, and shear rates between groups and treatments were compared with the one-way analysis of variance on ranks (Kruskal-Wallis) and with a multiple pairwise comparison test (Dunn test). Leukocyte counts and differentials were compared with the Student t test or by the Wilcoxon rank-sum test as appropriate. Statistical significance was set at P < .05.
All mice used in this work appeared healthy, active, and of normal
size and weight for their ages. The systemic leukocyte counts were
significantly higher in C2GlcNAcT-I-deficient mice than in wild-type
mice. This was true for both the group without pretreatment and the
TNF
Leukocyte rolling in untreated venules Leukocyte rolling was analyzed in 62 venules of 11 core 2/ mice and compared with rolling in 25 venules of 6 littermate controls. Hemodynamic parameters for both groups are
presented in Table 2 and show no
significant differences in diameter, blood flow velocity, shear rate,
or systemic blood pressure. Leukocyte rolling was assessed as leukocyte
rolling flux fraction, which is defined as the number of rolling
leukocytes divided by the total number of leukocytes passing through
the same vessel.30 During the first hour after
exteriorization of the cremaster muscle, rolling flux fraction
was only 4% in C2GlcNAcT-I-deficient mice, compared with 27%
in control mice (P < .05) (Figure
1A). Leukocyte rolling during that time
was almost exclusively P-selectin dependent, because it was blocked by
the P-selectin mAb RB40.34 (Figure 1A). The lower leukocyte rolling
flux fraction in core 2/ mice can therefore be
attributed to a severe impairment in P-selectin-mediated rolling.
To investigate whether PSGL-1 was necessary for P-selectin-dependent
rolling in core 2 Average rolling velocity (Vavg) in core 2 To show that the PSGL-1-blocking mAb 4RA10 (30 µg) was injected at a
saturating dose, 4 control mice were treated with increasing doses (1 µg, 3 µg, 10 µg, and 100 µg) of the PSGL-1-blocking mAb 4RA10.
Fluorescence-activated cell sorter (FACS) analysis of blood obtained 20 minutes after injection showed that 10 µg per mouse saturated all
binding sites on neutrophils, and no further increase was seen at 100 µg 4RA10 (Figure 2). In addition,
rolling flux fraction was assessed before and after injection of 30 µg 4RA10 followed by additional injection of 70 µg 4RA10, for a
total dose of 100 µg 4RA10. The additional injection led to no
further drop in rolling flux fraction. Subsequent injection of mAb
RB40.34 abolished rolling completely. This confirms that the dose of 30 µg 4RA10 per mouse was also functionally saturating.
To verify that PSGL-1 was indeed significantly modified by the
C2GlcNAcT-I enzyme, we probed bone marrow neutrophils and their precursors for PSGL-1 protein. We found an increased mobility of PSGL-1
from core 2
Short-term (2-hour) TNF for 2 hours induces the expression of
E-selectin and enhances the expression of P-selectin on venules of the
cremaster muscle.21 We assessed leukocyte rolling in 55 venules of 11 TNF-treated mice lacking C2GlcNAcT-I and compared the
results with rolling in 43 venules of 11 control animals. Hemodynamic
parameters for both groups are presented in Table 2 and show similar
vessel diameters, centerline velocities, wall shear rates, and
systemic blood pressures. Leukocyte rolling flux fraction
after TNF treatment was reduced to 8% in core 2/
mice compared with 21% in control mice (P < .05) (Figure
4), suggesting a severe defect in
selectin ligand function in C2GlcNAcT-I-deficient mice.
To investigate E-selectin-mediated rolling in core 2 Since the defect for E-selectin-mediated rolling in core
2
To study P-selectin-dependent rolling in core 2 Next, we investigated leukocyte rolling velocities and velocity
distributions in TNF
Leukocyte differentials in cremaster muscle venules To determine the effect of eliminating C2GlcNAcT-I on the composition of cells in venules (intravascular) and recruited into cremaster tissue (perivascular), we used Giemsa-stained whole-mount mouse cremaster muscles to differentiate leukocytes in TNF-treated mice. The number of intravascular neutrophils was significantly reduced
by about 90% in core 2/ mice compared with control
mice (Figure 7A). By contrast, adhesion of eosinophils or mononuclear cells appeared to be unaffected by the
absence of C2GlcNAcT-I. To directly address the contribution of
E-selectin to leukocyte accumulation in venules, we investigated the
number of intravascular leukocytes in cremaster muscle of mice treated
with mAbs to L- and P-selectin shortly before injection of TNF. This
treatment did not alter leukocyte adhesion, suggesting that the absence
of C2GlcNAcT-I causes a severe reduction in neutrophil adhesion that
cannot be achieved by blocking P- and L-selectin. Similar to the
findings on intravascular leukocytes, absence of C2GlcNAcT-I also
reduced the number of extravascular neutrophils, but not eosinophils or
mononuclear cells (Figure 7B).
In this study, we have used the mouse cremaster muscle to
investigate the role of C2GlcNAcT-I in the biosynthesis and
physiological function of selectin ligands in vivo. We found that in
cremaster muscle venules of C2GlcNAcT-I-deficient mice, P- and
E-selectin-dependent rolling is sharply reduced. In addition, we show
that the observed defect in E-selectin-mediated rolling becomes
evident only at wall shear rates above 300 s In untreated cremaster muscle venules of C2GlcNAcT-I-deficient but not
control mice, P-selectin-mediated rolling is completely blocked by a
mAb to PSGL-1. This is a novel finding, demonstrating that C2GlcNAcT-I
is required for optimal PSGL-1 function, but that PSGL-1 lacking core 2 oligosaccharides can still bind to P-selectin at a rate and affinity
that allow some leukocyte rolling. The significant size difference in
PSGL-1 monomer and dimer expressed on leukocytes of core
2 Previous in vitro work had suggested that PSGL-1 requires the core 2 modification to bind to P-selectin.15 CHO cells
transfected with PSGL-1 and fucosyltransferase IV required C2GlcNAcT-I
to avidly bind fluid-phase P-selectin. Kumar et al14 found
some residual binding of soluble P-selectin to PSGL-1 when
cotransfected with fucosyltransferase III but without C2GlcNAcT-I into
CHO cells. Binding was much improved by adding C2GlcNAcT-I.
Ramachandran et al33 showed that CHO cells transfected
with fucosyltransferase VII and PSGL-1 did not support rolling of a
pre-B cell line expressing P-selectin in a flow chamber system.
However, rolling was found after cotransfection with C2GlcNAcT-I. This
flow chamber assay is reversed in orientation compared with the in vivo
situation (P-selectin on the rolling cell, PSGL-1 on the substrate),
which could be of significance, since PSGL-1 is known to be
preferentially localized to the tips of microvilli on
leukocytes.34 Taken together, our findings confirm and
extend previous in vitro findings to physiologically relevant wall
shear rates. Interestingly, the velocity of rolling leukocytes after
blocking PSGL-1 was increased to a degree similar to that in
C2GlcNAcT-I-deficient mice. This shows that PSGL-1 in the absence of
C2GlcNAcT-I supports rolling only at a higher velocity, suggesting an
increased off-rate of the P-selectin-PSGL-1 bond in core
2 The present data also show that other ligands for P-selectin may exist
and mediate some leukocyte rolling in control mice, consistent with
previous findings31 and recent findings in
PSGL-1-deficient mice.35 However, there is a quantitative
difference between the degree of rolling after blocking PSGL-1. Under
conditions where a mAb to P-selectin blocked almost all leukocyte
rolling, a mAb to PSGL-1 blocked only 73% in control mice, although
P-selectin-dependent rolling was reduced by 95% in PSGL-1-deficient
mice.35 Therefore, we cannot formally exclude the
possibility that mAb 4RA10, even at saturating concentrations, may not
completely block PSGL-1 function in wild-type mice. Since
PSGL-1-blocking mAb 4RA10 completely blocked rolling in core
2 Requirements for E-selectin-dependent rolling in vivo are not well understood. Therefore, we addressed whether E-selectin ligands require modification by C2GlcNAcT-I. It is clear that E-selectin ligands must be modified by fucosyltransferase VII or IV or another fucosyltransferase in order to be functional.36 In fact, transfection of fucosyltransferase VII into all cell lines tested conferred the ability to bind to E-selectin in a flow chamber assay,37,38 suggesting that the glycoprotein requirements for E-selectin binding may be more relaxed than for P-selectin. Although PSGL-1 has been shown to bind to E-selectin,15,39,40 recent studies in PSGL-1-deficient mice show that PSGL-1 is not required for neutrophil rolling through E-selectin.35 Another candidate ligand for E-selectin, ESL-1,41 awaits confirmation of significance in vivo. ESL-1 is decorated with N-linked but not O-linked carbohydrates. Therefore, ESL-1 glycosylation should not be affected in C2GlcNAcT-I-deficient mice. The present data show that E-selectin-dependent rolling operates in
core 2 In conclusion, the present study has identified some of the mechanisms
by which the absence of C2GlcNAcT-I impairs neutrophil recruitment to
sites of inflammation.16 We conclusively show (1) that
P-selectin-dependent rolling via PSGL-1 is severely impaired, (2) that
rolling through other P-selectin ligands is completely absent in core
2
We thank Drs Ruth Eytner, Martin Wild, and Dietmar Vestweber, Universität Münster, Germany, for providing mAbs RB40.34 and 4RA10, and Dr Barry Wolitzky for providing mAb 9A9.
Submitted November 14, 2000; accepted February 13, 2001.
Supported by National Institutes of Health (NIH) grants HL-64381 and HL-54136 (K.L.); by NIH grant DK 48247 (J.D.M.); by a stipend from the German Research Foundation (DFG) SP 621/1-1 (M.S.); and by National Cancer Institute Fellowship F32CA79130 (L.G.E.).
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Klaus Ley, Department of Biomedical Engineering and Cardiovascular Research Center, University of Virginia, Health Sciences Center, Box 800759, Charlottesville, VA 22908; e-mail: klausley{at}virginia.edu.
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© 2001 by The American Society of Hematology.
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H. Wang, R. Tang, W. Zhang, K. Amirikian, Z. Geng, J. Geng, R. P. Hebbel, L. Xia, J. D. Marth, M. Fukuda, et al. Core2 1-6-N-Glucosaminyltransferase-I Is Crucial for the Formation of Atherosclerotic Lesions in Apolipoprotein E-Deficient Mice Arterioscler Thromb Vasc Biol, February 1, 2009; 29(2): 180 - 187. [Abstract] [Full Text] [PDF]
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 S. Yakubenia, D. Frommhold, D. Scholch, C. C. Hellbusch, C. Korner, B. Petri, C. Jones, U. Ipe, M. G. Bixel, R. Krempien, et al. Leukocyte trafficking in a mouse model for leukocyte adhesion deficiency II/congenital disorder of glycosylation IIc Blood, August 15, 2008; 112(4): 1472 - 1481. [Abstract] [Full Text] [PDF]
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 D. Frommhold, A. Ludwig, M. G. Bixel, A. Zarbock, I. Babushkina, M. Weissinger, S. Cauwenberghs, L. G. Ellies, J. D. Marth, A. G. Beck-Sickinger, et al. Sialyltransferase ST3Gal-IV controls CXCR2-mediated firm leukocyte arrest during inflammation J. Exp. Med., June 9, 2008; 205(6): 1435 - 1446. [Abstract] [Full Text] [PDF]
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B. Leon and C. Ardavin Monocyte migration to inflamed skin and lymph nodes is differentially controlled by L-selectin and PSGL-1 Blood, March 15, 2008; 111(6): 3126 - 3130. [Abstract] [Full Text] [PDF]
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J. Schymeinsky, A. Sindrilaru, D. Frommhold, M. Sperandio, R. Gerstl, C. Then, A. Mocsai, K. Scharffetter-Kochanek, and B. Walzog The Vav binding site of the non-receptor tyrosine kinase Syk at Tyr 348 is critical for beta2 integrin (CD11/CD18)-mediated neutrophil migration Blood, December 1, 2006; 108(12): 3919 - 3927. [Abstract] [Full Text] [PDF]
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D. A. Carlow and H. J. Ziltener CD43 Deficiency Has No Impact in Competitive In Vivo Assays of Neutrophil or Activated T Cell Recruitment Efficiency J. Immunol., November 1, 2006; 177(9): 6450 - 6459. [Abstract] [Full Text] [PDF]
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G. H. Underhill, D. G. Zisoulis, K. P. Kolli, L. G. Ellies, J. D. Marth, and G. S. Kansas A crucial role for T-bet in selectin ligand expression in T helper 1 (Th1) cells Blood, December 1, 2005; 106(12): 3867 - 3873. [Abstract] [Full Text] [PDF]
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 M. A. V. Landgraf, L. L. Martinez, V. M. F. Rastelli, M. d. C. P. Franco, M. Soto-Suazo, R. d. C. A. Tostes, M. H. C. Carvalho, D. Nigro, and Z. B. Fortes Intrauterine Undernutrition in Rats Interferes with Leukocyte Migration, Decreasing Adhesion Molecule Expression in Leukocytes and Endothelial Cells J. Nutr., June 1, 2005; 135(6): 1480 - 1485. [Abstract] [Full Text] [PDF]
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J. S. Merzaban, J. Zuccolo, S. Y. Corbel, M. J. Williams, and H. J. Ziltener An Alternate Core 2 {beta}1,6-N-Acetylglucosaminyltransferase Selectively Contributes to P-Selectin Ligand Formation in Activated CD8 T Cells J. Immunol., April 1, 2005; 174(7): 4051 - 4059. [Abstract] [Full Text] [PDF]
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U. Syrbe, S. Jennrich, A. Schottelius, A. Richter, A. Radbruch, and A. Hamann Differential regulation of P-selectin ligand expression in naive versus memory CD4+ T cells: evidence for epigenetic regulation of involved glycosyltransferase genes Blood, November 15, 2004; 104(10): 3243 - 3248. [Abstract] [Full Text] [PDF]
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M. J. Smith, B. R. E. Smith, M. B. Lawrence, and K. R. Snapp Functional Analysis of the Combined Role of the O-Linked Branching Enzyme Core 2 {beta}1-6-N-Glucosaminyltransferase and Dimerization of P-selectin Glycoprotein Ligand-1 in Rolling on P-selectin J. Biol. Chem., May 21, 2004; 279(21): 21984 - 21991. [Abstract] [Full Text] [PDF]
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T.-Y. Yen, B. A. Macher, S. Bryson, X. Chang, I. Tvaroska, R. Tse, S. Takeshita, A. M. Lew, and A. Datti Highly Conserved Cysteines of Mouse Core 2 {beta}1,6-N-Acetylglucosaminyltransferase I Form a Network of Disulfide Bonds and Include a Thiol That Affects Enzyme Activity J. Biol. Chem., November 14, 2003; 278(46): 45864 - 45881. [Abstract] [Full Text] [PDF]
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S. M. Barry, D. G. Zisoulis, J. W. Neal, N. A. Clipstone, and G. S. Kansas Induction of FucT-VII by the Ras/MAP kinase cascade in Jurkat T cells Blood, September 1, 2003; 102(5): 1771 - 1778. [Abstract] [Full Text] [PDF]
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Y.-C. Lim, G. Garcia-Cardena, J. R. Allport, M. Zervoglos, A. J. Connolly, M. A. Gimbrone Jr., and F. W. Luscinskas Heterogeneity of Endothelial Cells from Different Organ Sites in T-Cell Subset Recruitment Am. J. Pathol., May 1, 2003; 162(5): 1591 - 1601. [Abstract] [Full Text] [PDF]
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A. E. R. Hicks, S. L. Nolan, V. C. Ridger, P. G. Hellewell, and K. E. Norman Recombinant P-selectin glycoprotein ligand-1 directly inhibits leukocyte rolling by all 3 selectins in vivo: complete inhibition of rolling is not required for anti-inflammatory effect Blood, April 15, 2003; 101(8): 3249 - 3256. [Abstract] [Full Text] [PDF]
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C. J. Dimitroff, R. J. Bernacki, and R. Sackstein Glycosylation-dependent inhibition of cutaneous lymphocyte-associated antigen expression: implications in modulating lymphocyte migration to skin Blood, January 15, 2003; 101(2): 602 - 610. [Abstract] [Full Text] [PDF]
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M. P. Bernimoulin, X.-L. Zeng, C. Abbal, S. Giraud, M. Martinez, O. Michielin, M. Schapira, and O. Spertini Molecular Basis of Leukocyte Rolling on PSGL-1. PREDOMINANT ROLE OF CORE-2 O-GLYCANS AND OF TYROSINE SULFATE RESIDUE 51 J. Biol. Chem., January 3, 2003; 278(1): 37 - 47. [Abstract] [Full Text] [PDF]
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M. J. Martin, T. Feizi, C. Leteux, D. Pavlovic, V. E. Piskarev, and W. Chai An investigation of the interactions of E-selectin with fuco-oligosaccharides of the blood group family Glycobiology, December 1, 2002; 12(12): 829 - 835. [Abstract] [Full Text] [PDF]
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E. E. Burch, V. R. S. Patil, R. T. Camphausen, M. F. Kiani, and D. J. Goetz The N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated endothelium via P-selectin in vivo Blood, June 28, 2002; 100(2): 531 - 538. [Abstract] [Full Text] [PDF]
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Y.-C. Lim, H. Xie, C. E. Come, S. I. Alexander, M. J. Grusby, A. H. Lichtman, and F. W. Luscinskas IL-12, STAT4-Dependent Up-Regulation of CD4+ T Cell Core 2 {beta}-1,6-n-Acetylglucosaminyltransferase, an Enzyme Essential for Biosynthesis of P-Selectin Ligands J. Immunol., October 15, 2001; 167(8): 4476 - 4484. [Abstract] [Full Text] [PDF]
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K. R. Snapp, C. E. Heitzig, L. G. Ellies, J. D. Marth, and G. S. Kansas Differential requirements for the O-linked branching enzyme core 2 {beta}1-6-N-glucosaminyltransferase in biosynthesis of ligands for E-selectin and P-selectin Blood, June 15, 2001; 97(12): 3806 - 3811. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
) and homozygous (+/+)
littermates. All experiments were performed on healthy mice that were
at least 8 weeks of age. Mice were housed in a barrier facility under
specific-pathogen-free conditions, and animal experiments were
approved by the animal care and use committee of the University of Virginia.
) and control mice (
). * indicates
significant differences (P < .05) in leukocyte rolling
flux fraction between core 2
) and core 2