The JI
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     
 

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Matsumoto, M.
Right arrow Articles by Hirata, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Matsumoto, M.
Right arrow Articles by Hirata, T.
The Journal of Immunology, 2007, 178: 2499-2506.
Copyright © 2007 by The American Association of Immunologists, Inc.

CD43 Collaborates with P-Selectin Glycoprotein Ligand-1 to Mediate E-Selectin-Dependent T Cell Migration into Inflamed Skin1

Masanori Matsumoto*,{dagger}, Akiko Shigeta*, Yuko Furukawa*,{dagger}, Toshiyuki Tanaka{dagger}, Masayuki Miyasaka{dagger} and Takako Hirata2,*

* The 21st Century Center of Excellence Program, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan; and {dagger} Laboratory of Immunodynamics, Department of Microbiology and Immunology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 Disclosures
 References
 
Activated T cell migration into nonlymphoid tissues is initiated by the interactions of P- and E-selectin expressed on endothelial cells and their ligands on T cells. P-selectin glycoprotein ligand-1 (PSGL-1) has been the only E-selectin ligand demonstrated to function during the in vivo migration of activated T cells. We show in this study that CD43-deficient Th1 cells, like PSGL-1-deficient cells, exhibited reduced E-selectin-binding activity compared with wild-type cells. Th1 cells with a PSGL-1 and CD43 double deficiency showed even less E-selectin-binding activity. In migration assays in which adoptively transferred cells migrate to inflamed skin P- and E-selectin dependently, CD43 contributed significantly to PSGL-1-independent Th1 cell migration. In addition, in vivo activated T cells from the draining lymph nodes of sensitized mice deficient in PSGL-1 and/or CD43 showed significantly decreased E-selectin-binding activity and migration efficiency, with T cells from double-deficient mice showing the most profound decrease. Collectively, these results demonstrate that the CD43 expressed on activated T cells functions as an E-selectin ligand and thereby mediates T cell migration to inflamed sites, in collaboration with PSGL-1.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 Disclosures
 References
 
Naive T cells traffic through secondary lymphoid organs, such as the lymph nodes (LNs)3, Peyer’s patches, and spleen, where they are most likely to encounter their cognate Ag. Following an Ag encounter, naive T cells undergo clonal expansion and differentiate into effector/memory T cells. These T cells leave the secondary lymphoid organs and migrate into nonlymphoid tissues, such as the skin, where they carry out their specific functions (1, 2). The migration of effector/memory T cells to the skin is crucial for successful immune surveillance, as the skin constitutes the primary barrier against trauma and a broad array of microbial pathogens. The presence of T cells in the skin also plays a key role in the pathogenesis of many inflammatory skin diseases, such as atopic dermatitis, allergic contact dermatitis, and psoriasis (3).

A dynamic multistep process is thought to be required for the T cells to exit the bloodstream and enter the skin, and this process is involved in both constitutive memory T cell trafficking and inflammation-induced effector T cell recruitment (4). This multistep process is initiated by a tethering and rolling step that captures T cells from the bloodstream and allows them to roll on the surface of endothelial cells under blood flow. These interactions are primarily mediated by selectins and their ligands. Skin endothelium constitutively expresses both P-selectin (CD62P) and E-selectin (CD62E) (5), and the expression of both selectins is up-regulated during infection and in many inflammatory conditions of the skin (6, 7, 8), which promotes the recruitment of T cells that express P- and E-selectin ligands.

The major ligand for P-selectin on effector/memory T cells is P-selectin glycoprotein ligand-1 (PSGL-1; CD162), a sialomucin expressed on most leukocytes (9, 10). Th1 cells deficient in PSGL-1 do not bind to P-selectin nor do they migrate into the inflamed skin of E-selectin-deficient mice in a contact hypersensitivity (CHS) model, in which the migration is largely dependent on P-selectin, indicating that PSGL-1 plays a critical role as a P-selectin ligand (10). In contrast, the migration of PSGL-1-deficient (PSGL-1–/–) Th1 cells into the inflamed skin of P-selectin-deficient mice is reduced, but not abolished, suggesting that although PSGL-1 is an E-selectin ligand, it is not the only one (10). It is not yet known what physiological E-selectin ligands besides PSGL-1 are expressed on Th1 cells.

E-selectin recognizes sialylated and fucosylated carbohydrate structures such as sialyl LewisX (sLeX) (11). The presentation of such carbohydrates on the protein backbone contributes to the specificity and affinity of the selectin binding. Besides PSGL-1, several glycoproteins that bind E-selectin have been reported. E-selectin ligand-1 (ESL-1) is a transmembrane glycoprotein that was identified using a recombinant E-selectin-IgG chimera as the major E-selectin ligand on mouse neutrophils (12). In contrast to PSGL-1, which carries sLeX on O-linked glycans, the specific glycoform of ESL-1 expressed on myeloid cells carries sLeX on N-linked glycans. L-selectin from human neutrophils and cultured T lymphoblasts, but not from mouse cells, is capable of binding E-selectin (13, 14, 15, 16). However, whether these glycoprotein ligands support physiologically relevant interactions with E-selectin is not yet certain. Recently, CD44, a hyaluronan-binding cell surface glycoprotein, was reported to bind E-selectin through N-linked glycans and to mediate the E-selectin-dependent rolling of neutrophils (17). However, whether CD44 mediates T cell interactions with E-selectin has not been studied.

We recently showed that the 130-kDa glycoform of CD43, expressed on mouse Th1 cells, binds E-selectin (18). In humans, CD43 on activated T cells also binds E-selectin (18, 19), and human CD43 is decorated with the cutaneous lymphocyte-associated Ag, which is a carbohydrate epitope expressed on skin-homing effector/memory T cells (19). These findings raise the possibility that CD43 mediates E-selectin-dependent processes in vivo. Although CD43 is abundant on T cells, its role remains elusive, and it is implicated in both antiadhesive and proadhesive roles (20). Targeted disruption of CD43 in cell lines and mice leads to increased T cell adhesion and proliferation (21, 22) as well as increased T cell migration to secondary lymphoid organs in vivo (23), implicating CD43 in an antiadhesive process. However, the administration of an anti-CD43 mAb inhibits T cell migration into secondary lymphoid organs (24). The same mAb blocks the migration of T cells into inflamed pancreatic islets and prevents diabetes in NOD mice (25). In addition, in a lymphocytic choriomeningitis virus (LCMV) infection model as well as in an experimental autoimmune encephalitis model, Ag-specific T cells exhibit significantly reduced migration into tissues such as the brain in CD43-deficient (CD43–/–) mice (26, 27). These findings are consistent with the idea that CD43 functions as a proadhesive molecule in T cell trafficking. There is no evidence, however, that the proadhesive function of CD43 observed in vivo is related to its function as a selectin ligand.

To determine the role of CD43 as an E-selectin ligand in vivo and the relative contribution of PSGL-1 and CD43 to E-selectin-mediated T cell migration, we generated mice deficient in both PSGL-1 and CD43. Using Th1 cells from PSGL-1–/–, CD43–/–, and PSGL-1/CD43 double-deficient (DKO) mice, we analyzed the role of CD43 as an E-selectin ligand in comparison with PSGL-1. We show that DKO Th1 cells exhibited a more profound reduction in E-selectin binding and migration into the inflamed skin in a CHS model than PSGL-1–/– Th1 cells. In addition, we show that LN T cells from sensitized mice deficient in PSGL-1 and/or CD43 migrated into the inflamed skin less efficiently than those from wild-type (WT) mice. These results indicate that CD43 functions as an E-selectin ligand and collaborates with PSGL-1 to mediate T cell migration in vivo.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 Disclosures
 References
 
Mice

C57BL/6 (B6) mice were purchased from CLEA Japan. PSGL-1–/– mice on a B6 background were provided by B. Furie (Harvard Medical School, Boston, MA). CD43–/– mice on a B6 x 129S4/SvJae background were purchased from The Jackson Laboratory. PSGL-1–/– mice were intercrossed with CD43–/– mice to generate double heterozygotes. These doubly heterozygous mice were then bred to yield WT, PSGL-1–/–, CD43–/–, and DKO mice. All the mice used were 6–10 wk old. The mice were housed at the Institute of Experimental Animal Sciences at Osaka University Medical School. All studies and procedures were approved by the Ethics Review Committee for Animal Experimentation of the Osaka University Graduate School of Medicine.

Chimeric proteins

The expression plasmids for mouse P- and E-selectin-IgM chimeric proteins were provided by J. Lowe (University of Michigan Medical School, Ann Arbor, MI). COS-7 cells were transfected with the plasmids using DEAE-dextran. Mouse P- and E-selectin-IgG chimeric proteins were prepared, as described previously (10).

Preparation of mouse Th1 cells

Splenic CD4+ T cells were isolated with a BD IMag CD4 T Lymphocyte Enrichment Set (BD Biosciences). The enriched population was >90% positive for CD4 staining. Purified CD4+ T cells were cultured on 6-cm tissue culture dishes coated with 10 µg/ml anti-CD3{epsilon} (145-2C11; BD Biosciences) and 10 µg/ml anti-CD28 (37.51; BD Biosciences) for 2 days in the presence of 4 ng/ml IL-2 (R&D Systems), 8 ng/ml IL-12 (R&D Systems), and 0.2 µg/ml anti-IL-4 (11B11; BD Biosciences). The cells were then transferred to uncoated dishes and cultured for an additional 4 days.

Flow cytometry

To assess the selectin-IgM binding, cells were incubated with a COS-7 supernatant containing P-selectin-IgM, E-selectin-IgM, or control human IgM. Nonspecific staining was determined by the addition of 5 mM EDTA. The cells were washed and incubated with biotinylated anti-human IgM (American Qualex). The cells were then washed; stained with streptavidin (SA)-PE (BD Biosciences) alone or together with anti-CD44-PE-Cy5 (IM7; eBioscience), anti-CD45RB-FITC (16A; BD Biosciences), and anti-CD4-allophycocyanin (RM4-5; Biolegend); and analyzed on a FACSCalibur (BD Biosciences).

Cell adhesion assays

The cell adhesion assays were performed, as previously described (10). In brief, P- or E-selectin-IgG or control human IgG (Sigma-Aldrich) (10 µg/ml) was immobilized on 96-well plates (Sumilon H; Sumitomo Bakelite) at 4°C overnight, and the plates were blocked with 1% BSA in PBS at 37°C for 2 h. Th1 cells resuspended in HBSS containing 2 mM CaCl2 or 5 mM EDTA were added to the plate and incubated for 20 min at 4°C with rotation (100 rpm). After the plates were washed three times with HBSS containing 2 mM CaCl2 or 5 mM EDTA, the number of bound cells was determined by photographing the cells and counting them.

Precipitation with E-selectin-IgG

WT, PSGL-1–/–, CD43–/–, or DKO Th1 cells were washed three times with PBS and surface biotinylated in PBS containing 0.5 mg/ml sulfo-normal human serum-Langerhans cells-biotin (Pierce) (2.5 x 107 cells/ml) at room temperature for 30 min. The cells were then washed three times with PBS and lysed at a density of 3 x 107 cells/ml in cold lysis buffer (1% Triton X-100, 50 mM Tris (pH 7.4), 150 mM NaCl, 1 mM CaCl2, and 1 mM PMSF) for 30 min. Insoluble materials were pelleted at 15,000 x g for 20 min. The supernatant was aliquoted, and a fraction corresponding to 1 x 107 cells was incubated for 4 h with 50 µl of packed protein A-Sepharose. After removal of the Sepharose beads, the lysate was incubated in the presence of 1 mM CaCl2 with 20 µl of protein A-Sepharose preloaded for 4 h at 4°C with 50 µg of E-selectin-IgG or human IgG. After a 4-h incubation, the beads were washed five times with wash buffer (1% Triton X-100, 50 mM Tris (pH 7.4), 150 mM NaCl, and 1 mM CaCl2). Proteins bound to E-selectin-IgG were eluted with elution buffer (5 mM EDTA, 50 mM Tris (pH 7.4), and 0.05% Triton X-100). Eluted materials were separated by SDS-PAGE under nonreducing conditions and transferred to an Immobilon-P membrane (Millipore). Membranes were blotted with HRP-conjugated SA (SA-HRP; Zymed Laboratories). The membranes were also blotted with a polyclonal anti-mouse PSGL-1 Ab (28), an anti-mouse CD43 mAb 1B11 (BD Biosciences), or a polyclonal anti-mouse ESL-1 Ab (provided by B. Furie, Harvard Medical School, Boston, MA), followed by the appropriate HRP-conjugated secondary Abs (all from American Qualex).

Induction of CHS

Mice were sensitized by the application of 100 µl of 2% (w/v) oxazolone (Sigma-Aldrich) in 4:1 acetone/olive oil on the shaved abdominal skin on day 0. Some mice were painted with acetone/olive oil alone. The mice were challenged on day 6 by applying 20 µl of 0.5% (w/v) oxazolone to the left ear (10 µl per side). The right ear was painted with the vehicle only. Ear-swelling responses were measured using a dial thickness gauge (Mitutoyo).

Isolation of LN cells

Mice were sensitized with oxazolone on the shaved abdomen on day 0. On day 6, the mice were killed, their draining LNs were removed, and the T cells were isolated using a BD IMag T Lymphocyte Enrichment Set (BD Biosciences). The cells were used for flow cytometric analyses or as donor cells in migration assays. For LN total cell counts, mice were sensitized by the application of 20 µl of 2% (w/v) oxazolone to the left ear (10 µl per side). The right ear was painted with the vehicle only. On day 6, the mice were killed, the cervical LNs of both sides were removed, and the nucleated cells were counted on a hemocytometer.

In vivo migration assays

Th1 cells were harvested after 6 days of culture, and dead cells were removed by centrifugation on Ficoll-PaquePLUS (GE Healthcare). T cells were isolated from draining LNs from oxazolone-sensitized mice. The cells (1~2 x 107/ml) were radiolabeled with 50 µCi/ml sodium [51Cr]chromate (MP Biomedicals) for 1 h at 37°C, washed twice, and resuspended in PBS. Th1 cells (5 x 106) or LN T cells (2 x 107) were injected into the tail veins of mice that had been sensitized 7 days before and challenged on the left ear 24 h before. In some experiments, the anti-P-selectin mAb RB40.34 (BD Biosciences), the anti-E-selectin mAb 9A9 provided by B. Wolitzky (Coelacanth, East Windsor, NJ), or the isotype control (50 µg/mouse) was injected together with the cells. The mice were killed 3 h (Th1 cells) or 24 h (LN T cells) after injection, and the radioactivity in the ear and spleen was measured using a Packard Instrument gamma scintillation counter.

Histologic analyses

Ear specimens were taken 24 h after the challenge. Histological analyses were performed on formalin-fixed, paraffin-embedded sections stained with H&E.

Isolation of skin-infiltrating cells

Skin-infiltrating cells were isolated via enzyme digestion. Briefly, ears taken 24 h after the challenge were separated into ventral and dorsal sheets. The sheets were cut into small pieces and incubated in RPMI 1640 containing 10% FCS, 400 U/ml collagenase (Roche), and 10 µg/ml DNase I (Roche), with continuous stirring at 37°C for 90 min. The resulting cell population was filtered through a 100-µm strainer (BD Falcon) and then enriched for lymphocytes by centrifugation on Ficoll-PaquePLUS. The cells were stained with anti-CD44-FITC (IM7; eBioscience), anti-CD45RB-PE (16A; BD Biosciences), and anti-CD4-allophycocyanin, and analyzed on a FACSCalibur.

Statistical analysis

Data are presented as the mean ± SEM. Statistical analyses were performed using the two-tailed unpaired Student’s t test.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 Disclosures
 References
 
Th1 cells deficient in both PSGL-1 and CD43 show a more profound decrease in E-selectin binding than cells deficient in either molecule alone

To investigate the role of CD43 as an E-selectin ligand, we first tested Th1 cells generated in vitro from CD43–/– mice for selectin-binding activities. As reported previously (18), flow cytometric assays using selectin-IgM chimeric proteins showed that Th1 cells from PSGL-1–/– mice did not bind P-selectin-IgM at all and bound E-selectin-IgM slightly less well than WT cells did (Fig. 1A). CD43–/– Th1 cells also showed slightly reduced E-selectin-IgM binding, although they bound P-selectin-IgM about as well as WT cells (Fig. 1A). E-selectin-binding activity was also tested in cell adhesion assays using selectin-IgG chimeric proteins immobilized on plastic plates. The adhesion of CD43–/– Th1 cells to the immobilized E-selectin-IgG was reduced, as was that of PSGL-1–/– cells, by 48%, compared with WT cells (Fig. 1B). Consistent with the results of the flow cytometric analyses shown in Fig. 1A, the adhesion of PSGL-1–/– but not CD43–/– Th1 cells to P-selectin-IgG was completely abolished (Fig. 1B). These results confirm that PSGL-1 is the predominant P-selectin ligand on Th1 cells and indicate that both PSGL-1 and CD43 function as E-selectin ligands.


Figure 1
View larger version (40K):
[in this window]
[in a new window]

  		FIGURE 1. E-selectin-binding activities of Th1 cells generated from WT, PSGL-1–/–, CD43–/–, and DKO mice. A, P- and E-selectin-IgM binding of Th1 cells from WT, PSGL-1–/–, CD43–/–, and DKO mice. Th1 cells of each genotype were incubated with P- and E-selectin-IgM in the presence of calcium (open histograms) or EDTA (shaded histograms). Bound selectin-IgM was detected using biotinylated anti-human IgM and SA-PE. B, Adhesion of WT, PSGL-1–/–, CD43–/–, and DKO Th1 cells to P- and E-selectin-IgG immobilized on 96-well plates. Cells were added to 96-well plates coated with P-selectin-IgG, E-selectin-IgG, or human IgG, in the presence of calcium. The plates were rotated for 20 min, unbound cells were removed, and the number of bound cells was determined. Values are means ± SEM from triplicate wells. One of three similar, independent experiments is shown.

 
To examine the cooperative and relative roles of PSGL-1 and CD43 in E-selectin-mediated processes, we generated mice deficient in both PSGL-1 and CD43 by intercrossing PSGL-1–/– and CD43–/– mice. We confirmed the absence of cell surface expression of PSGL-1 and CD43 on T cells, B cells, and granulocytes in the DKO mice (data not shown). These mice developed normally, were fully fertile, and showed no signs of infection. Th1 cells generated from DKO mice bound E-selectin-IgM much less well than cells deficient in either molecule alone (Fig. 1A). Similarly, in cell adhesion assays, the DKO Th1 cells showed a 73% reduction compared with WT cells, which was a more profound effect than that seen with either deficiency alone (Fig. 1B). The addition of EDTA completely abolished the adhesion of cells from all of the genotypes (data not shown), confirming that the adhesion was calcium dependent. These results indicate that both PSGL-1 and CD43 contribute to E-selectin-binding activity and that the loss of both molecules has an additive effect on the reduction in E-selectin-binding activity.

The 130-kDa protein is not precipitated with E-selectin-IgG from CD43-deficient Th1 cells

We reported previously that, under nonreducing conditions, three bands, of ~130, 150, and 270 kDa, were precipitated from WT Th1 cells with E-selectin-IgG in the presence of calcium (18). The 270-kDa band was not detected in the E-selectin-IgG precipitate from PSGL-1–/– Th1 cells, indicating that this band represents PSGL-1 (18). When the precipitation experiments were performed using CD43–/– Th1 cells, the 150- and 270-kDa bands, but not the 130-kDa band, were detected (Fig. 2A), confirming the identity of the 130-kDa band as CD43. The identity of these bands was also confirmed using a polyclonal anti-PSGL-1 Ab (Fig. 2B) and an anti-CD43 mAb 1B11 (Fig. 2C). As expected, both of the 130- and 270-kDa bands, representing CD43 and PSGL-1, respectively, were absent in the E-selectin-IgG precipitate from DKO Th1 cells, whereas the 150-kDa band remained (Fig. 2A). A polyclonal anti-ESL-1 Ab detected a 150-kDa band in the E-selectin-IgG precipitate from Th1 cells of all four genotypes (Fig. 2D). These biochemical analyses confirm that CD43 is one of the E-selectin-binding proteins expressed on Th1 cells.


Figure 2
View larger version (12K):
[in this window]
[in a new window]

  		FIGURE 2. Precipitation of E-selectin-binding molecules with E-selectin-IgG from Th1 cells. WT, PSGL-1–/–, CD43–/–, and DKO Th1 cells were surface biotinylated, and their detergent extracts were incubated with control human IgG (lane 1) or E-selectin-IgG (lanes 2 and 3) bound to protein A-Sepharose in the presence of calcium (lanes 1 and 2) or EDTA (lane 3). Bound proteins were eluted with EDTA, separated by SDS-PAGE under nonreducing conditions, and subjected to Western blotting with SA-HRP (A), anti-PSGL-1 Ab (B), anti-CD43 mAb 1B11 (C), or anti-ESL-1 Ab (D). Arrowheads indicate the positions of three bands detected by SA-HRP. One of three similar, independent experiments is shown.

 
CD43 contributes significantly to PSGL-1-independent Th1 cell migration into inflamed skin

To study the role of CD43 in T cell migration in vivo, Th1 cells from WT, PSGL-1–/–, CD43–/–, and DKO mice were tested for their migration efficiency in an oxazolone-induced CHS model. Th1 cell migration into the inflamed skin in this model is dependent on P- and E-selectin (29). Donor Th1 cells of the four genotypes were radioactively labeled with chromium 51 and injected into WT recipient mice that had been sensitized to oxazolone applied to the abdomen 7 days before and challenged with oxazolone applied to the left ear 24 h before. The right ear was painted with vehicle and served as the noninflamed control. Three hours after the injection, the radioactivity of the cells that accumulated in the ears was measured. Confirming our previous results (10), the accumulation of PSGL-1–/– Th1 cells in the challenged skin was reduced by 60% compared with WT cells (Fig. 3A). In contrast, the accumulation of CD43–/– cells was not significantly different from that of WT cells. Importantly, DKO Th1 cells migrated significantly less efficiently than PSGL-1–/– Th1 cells into the inflamed skin (Fig. 3A). The migration into the control ear and spleen was comparable among the four genotypes (Fig. 3A and data not shown). These results indicate that CD43 contributes significantly to PSGL-1-independent migration of Th1 cells into inflamed skin.


Figure 3
View larger version (6K):
[in this window]
[in a new window]

  		FIGURE 3. Migration of Th1 cells into inflamed skin in a CHS model. A, Accumulation of chromium 51-labeled Th1 cells in the control and challenged ears. Th1 cells from WT, PSGL-1–/–, CD43–/–, and DKO mice were labeled with chromium 51 and injected into the tail veins of WT mice that had been sensitized 7 days before with oxazolone and challenged 24 h before on the left ear. The mice were killed 3 h after injection, and the radioactivity in the control and challenged ears was counted. Values are means ± SEM from four mice. B, Effect of an anti-P-selectin mAb on Th1 cell accumulation in the control and challenged ears. Th1 cells from WT, PSGL-1–/–, CD43–/–, and DKO mice were labeled with chromium 51 and injected with the anti-P-selectin mAb RB40.34 into sensitized and challenged WT mice. Values are means ± SEM from four mice. C, Effect of an anti-E-selectin mAb on Th1 cell accumulation in the control and challenged ears. Chromium 51-labeled Th1 cells from WT, PSGL-1–/–, CD43–/–, and DKO mice were injected with the anti-E-selectin mAb 9A9 into sensitized and challenged WT mice. Values are means ± SEM from four mice. Results represent one of three similar experiments. *, p < 0.05; **, p < 0.005; ***, p < 0.0005.

 
To examine the role of CD43 in a process specifically mediated by E-selectin, Th1 cells were injected along with the blocking anti-P-selectin mAb RB40.34, which allows E-selectin-mediated migration to be studied in isolation. Confirming our previous results (10), PSGL-1–/– Th1 cell migration was reduced by 26% relative to WT cells (Fig. 3B). CD43–/– cells also showed a slight reduction in migration efficiency, although this reduction did not reach statistical significance (Fig. 3B). These results suggest that CD43 plays only a small role in this E-selectin-mediated process or that the lack of CD43 can be compensated for by another E-selectin ligand, most likely PSGL-1. Consistent with the idea that PSGL-1 is the sole P-selectin ligand on Th1 cells, blocking the P-selectin function with RB40.34 did not affect the migration efficiency of PSGL-1–/– or DKO Th1 cells (Fig. 3, compare A with B), and thus DKO Th1 cells migrated less efficiently than PSGL-1–/– Th1 cells in this E-selectin-mediated process as well (Fig. 3B). These results indicate that CD43 acts as an E-selectin ligand during Th1 cell migration into the inflamed skin, although the contribution by CD43 appears relatively subtle when PSGL-1 is expressed normally. We also confirmed that CD43 does not play a significant role in an E-selectin-independent process of Th1 cell migration, because injecting the anti-E-selectin mAb 9A9 reduced the migration efficiency of WT and CD43–/– Th1 cells to a similar level (Fig. 3C). Because this E-selectin-independent process is mostly P-selectin dependent (29), this result is compatible with the observation that CD43–/– Th1 cells bind P-selectin as well as WT cells (Fig. 1).

The residual migration of DKO Th1 cells into the challenged ear was almost completely abrogated by injecting the anti-E-selectin mAb 9A9 (Fig. 3C), but not an isotype control mAb (data not shown). This result is compatible with the in vitro data showing that DKO cells still bound E-selectin to some extent (Fig. 1) and that the 150-kDa protein was precipitated with E-selectin-IgG from DKO cells (Fig. 2A). These results suggest that, although PSGL-1 and CD43 collaborate to mediate T cell migration into inflamed skin, yet another molecule mediates the residual E-selectin-dependent migration.

LN T cells from sensitized mice deficient in PSGL-1 and/or CD43 show a reduction in E-selectin binding and migration into inflamed skin

The above results showed that CD43 serves as an E-selectin ligand on Th1 cells. Because these Th1 cells were generated in vitro, we next examined whether CD43 expressed on in vivo stimulated T cells functions as an E-selectin ligand. WT, PSGL-1–/–, CD43–/–, and DKO mice were sensitized with oxazolone, and 6 days later draining LNs were removed. Before the sensitization, LNs in the PSGL-1–/–, CD43–/–, and DKO mice tended to be slightly larger than in WT mice and to have relatively higher cell numbers (Fig. 4A), although no alterations were observed in the percentage of cells expressing CD3, CD4, CD8, or B220, or in the ratio of naive/memory cells among the four genotypes (data not shown). After sensitization, draining LNs were enlarged in proportion to their presensitization size in all four genotypes, so they were slightly larger in the PSGL-1–/–, CD43–/–, and DKO mice and contained more cells, compared with those in WT mice (Fig. 4A).


Figure 4
View larger version (46K):
[in this window]
[in a new window]

  		FIGURE 4. E-selectin-binding activity of T cells from the draining LNs of oxazolone-sensitized mice. A, Cell counts in the draining LNs. WT, PSGL-1–/–, CD43–/–, and DKO mice were sensitized with oxazolone on the left ear, and 6 days later the draining left cervical LNs were removed. The right ear was painted with vehicle, and the right cervical LNs served as controls. The number of cells in each LN was counted on a hemocytometer. Values are means ± SEM from three mice. B–D, WT, PSGL-1–/–, CD43–/–, and DKO mice were sensitized with oxazolone on the abdomen, and 6 days later their draining LNs were removed. T cells were isolated from the draining LNs and incubated with P- and E-selectin-IgM in the presence of calcium or EDTA. The cells were then incubated with biotinylated anti-human IgM, followed by SA-PE, anti-CD45RB-FITC, anti-CD44-PE-Cy5, and anti-CD4-allophycocyanin, and analyzed by flow cytometry. P- and E-selectin-IgM binding of the total T cell population (B), percentage of CD4+ T cells that were CD4+CD44highCD45RBlow (C), and P- and E-selectin-IgM binding of the CD4+CD44highCD45RBlow population (D) are shown. B and D, Percentages of selectin-binding cells are given. The numbers in parentheses indicate geometric mean fluorescence intensity values of E-selectin-IgM-binding cells. Results represent one of three similar experiments.

 
T cells were isolated from draining LNs, and the selectin-binding activity of these cells was tested by flow cytometry using P- and E-selectin-IgM. Both P- and E-selectin-binding cells were enriched in the CD44high population (Fig. 4B). As expected, P-selectin-binding activity was entirely dependent on PSGL-1: almost no cells from PSGL-1–/– and DKO mice bound P-selectin (Fig. 4B). In contrast, the difference in the frequency of E-selectin-binding cells was quite subtle among the four genotypes when all T cells were analyzed (Fig. 4B), although the loss of PSGL-1 and CD43 showed a trend toward a gene dose-dependent decrease in the geometric mean fluorescence intensity of E-selectin-binding cells (Fig. 4B, numbers in parentheses). Because E-selectin-binding cells are enriched in the memory cell population, as defined by the CD44highCD45RBlow phenotype, we next analyzed the E-selectin-binding activity gating on CD44highCD45RBlow cells. The percentage of CD4+ and CD8+ T cells or those with a memory phenotype was not significantly different among the four genotypes (Fig. 4C and data not shown). The frequency of E-selectin-binding cells in the memory CD4+ T cell population was decreased in PSGL-1–/– and CD43–/– mice, and further decreased in DKO mice (Fig. 4D). The geometric mean fluorescence intensity of E-selectin-binding cells in this population was also slightly decreased in cells from PSGL-1–/– and CD43–/– mice, and further decreased in those from DKO mice (Fig. 4D, numbers in parentheses). These results indicate that PSGL-1 and CD43 both contribute to the E-selectin-binding activity of in vivo activated T cells.

Next, LN T cells isolated from sensitized WT, PSGL-1–/–, CD43–/–, and DKO mice were labeled with chromium 51 and adoptively transferred to oxazolone-sensitized and -challenged WT recipient mice. In this model, transferred cells migrate into the inflamed skin only if they express P- and/or E-selectin ligands (30). WT T cells accumulated in the challenged ear, but not in the control ear (Fig. 5). The migration of PSGL-1–/– and CD43–/– T cells into the challenged ear was significantly decreased, by 57 and 36%, respectively, compared with WT cells. DKO T cells exhibited the most prominent decrease in accumulation (76%). In contrast, the accumulation of injected T cells in other organs such as the spleen did not vary among the four genotypes (data not shown). These results indicate that CD43 is an E-selectin ligand on in vivo activated T cells and cooperates with PSGL-1 to mediate the migration of these cells into inflamed skin.


Figure 5
View larger version (3K):
[in this window]
[in a new window]

  		FIGURE 5. Migration of in vivo activated T cells to the inflamed skin in a CHS model. LN T cells isolated from the draining LNs of oxazolone-sensitized WT, PSGL-1–/–, CD43–/–, and DKO mice were labeled with chromium 51 and injected into WT mice previously sensitized with oxazolone and challenged on the left ear. The mice were killed 24 h after the injection, and the radioactivity in the control and challenged ears was counted. Values are means ± SEM from four mice. Results represent one of three similar experiments. *, p < 0.05; **, p < 0.005; ***, p < 0.0005.

 
Mice deficient in PSGL-1 and CD43 exhibit reduced T cell infiltration into inflamed skin

The observation that adoptively transferred DKO T cells migrated into inflamed skin with reduced efficiency suggested that DKO mice would have reduced CHS responses. Therefore, the CHS responses were examined in WT, PSGL-1–/–, CD43–/–, and DKO mice. The mice were sensitized with oxazolone and challenged 6 days later on the left ear. The kinetics of ear swelling was not significantly different among the four genotypes (data not shown). Histological analysis of ear skin sections 24 h after the challenge showed characteristic signs of inflammation, including interstitial edema and infiltrated cells in the dermis, in all four genotypes (Fig. 6A). The number of cells infiltrating the dermis, which were mostly neutrophils, was reduced by 34% in the PSGL-1–/– mice and by 33% in the DKO mice (Fig. 6B).


Figure 6
View larger version (48K):
[in this window]
[in a new window]

  		FIGURE 6. CHS responses and cell infiltration in inflamed skin. WT, PSGL-1–/–, CD43–/–, and DKO mice were sensitized with oxazolone and challenged 6 days later on the left ear. A, H&E-stained sections from the challenged ears of WT, PSGL-1–/–, CD43–/–, and DKO mice. Ear specimens were taken 24 h after the challenge. Scale bar, 100 µm. B, The number of infiltrating cells in the dermis of the challenged ears. Data are means ± SEM from four mice. C, The number of skin-infiltrating CD4+ T cells. The skin-infiltrating cells were isolated from challenged ears. The total cells were counted using a hemocytometer. The isolated cells were stained with anti-CD44-FITC, anti-CD45RB-PE, and anti-CD4-allophycocyanin, and analyzed by flow cytometry. Data are means ± SEM from five mice. *, p < 0.05; **, p < 0.005.

 
To quantify the T cells infiltrating the inflamed skin, lymphocytes were isolated from the challenged skin and analyzed by flow cytometry. Almost all of the CD4+ T cells recovered from the inflamed skin were of the memory phenotype (data not shown). The absolute number of recovered CD4+ T cells was 59% lower in PSGL-1–/– mice than in WT mice, but the number was comparable to WT in CD43–/– mice. The greatest reduction of infiltrated CD4+ T cells (76% decrease), compared with WT, was observed in the DKO mice (Fig. 6C). These results confirm that CD43 collaborates with PSGL-1 to mediate CD4+ T cell migration to the inflamed skin during CHS responses. However, the reduced migration of CD4+ T cells to the inflamed skin of DKO mice did not result in a decrease in ear swelling.


   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
Disclosures
 References
 
The identities and physiological contributions of E-selectin ligands are not well characterized. In this study, we demonstrate that CD43, a sialomucin abundantly expressed on T cells, functions as an E-selectin ligand during activated T cell migration into inflamed skin in a CHS model. The role of CD43 as an E-selectin ligand is most apparent in the absence of PSGL-1, the major P-selectin ligand that also serves as an E-selectin ligand on activated T cells (10).

PSGL-1 and CD43 share many features: both are sialomucins with an extended extracellular structure, abundantly expressed on T cells, located on microvilli, and become selectin-binding forms when appropriately glycosylated upon T cell activation. Our data show that the impact of a PSGL-1 deficiency on Th1 cell migration into inflamed skin, which is mediated by P- and E-selectin, is marked, but a CD43 deficiency in the context of a normal PSGL-1 locus does not affect Th1 cell migration detectably. This may be attributable to the role of PSGL-1 as a ligand for both P- and E-selectin. In a process mediated solely by E-selectin, the PSGL-1 deficiency modestly decreased the migration efficiency. The CD43 deficiency also had a tendency to diminish the migration efficiency, but it wasn’t statistically significant. The role of CD43 in Th1 cell migration was most apparent in the absence of PSGL-1, as shown by the significant contribution of CD43 to PSGL-1-independent Th1 cell migration. These results demonstrate that CD43 collaborates with PSGL-1 during E-selectin-mediated Th1 cell migration.

The contribution of CD43 to E-selectin-binding activity and thereby to migration efficiency was even more prominent in LN T cells activated in vivo, indicating that CD43 indeed functions as an E-selectin ligand during immune responses. The CD43 deficiency decreased the E-selectin-binding activity of T cells obtained from the draining LNs of sensitized mice, and it diminished the migration of these cells to the inflamed skin. Both CD4+ Th1 and CD8+ type 1 cytotoxic T cells are crucial effector cells in CHS responses to oxazolone in B6 mice (31). Type 1 cytotoxic T cell migration into the inflamed skin in this model is also partly E-selectin dependent (32). Given that total LN T cells were used in our migration assays, the CD43 deficiency may have diminished not only CD4+, but also CD8+ T cell migration. Additional work will be required to clarify the role of CD43 in CD8+ T cell migration. Among CD4+ T cells, a population that negatively regulates CHS responses has been reported (33, 34). The observation that ear swelling is normal in DKO mice, despite the reduced CD4+ T cell infiltration into the inflamed ear, may suggest that PSGL-1 and CD43 are also involved in the infiltration of these regulatory CD4+ T cells.

The function of CD43 remains elusive (35). Although it is thought to function as an antiadhesive molecule, because of its extended structure and negative charge, a role as a proadhesive molecule has also been suggested. Several potential ligands for CD43 have been reported, including ICAM-1 (36), galectin-1 (37), and sialoadhesin (38), although it is unclear whether any of these potential ligands function in vivo. In this study, we showed that CD43 on activated T cells serves as a ligand for E-selectin during T cell migration in vivo. Interestingly, in a model of LCMV infection, LCMV-specific CD8+ T cells exhibit significantly reduced infiltration to the leptomeninges in CD43–/– mice (26). In an experimental autoimmune encephalitis model, CD43–/– mice show decreased CD4+ T cell infiltration into the brain and spinal cord (27). Given that P- and E-selectin are implicated in lymphocyte recruitment to the inflamed brain (39), it is possible that the function of CD43 as an E-selectin ligand may account for the T cell recruitment defect observed in the CD43–/– mice.

The ability of CD43 to bind E-selectin is regulated by cell-specific posttranslational modifications. E-selectin recognizes sialylated and fucosylated carbohydrate structures such as sLeX, which are generated by the sequential action of several glycosyltransferases such as core 2 beta-1,6-N-acetylglucosaminyltransferase and {alpha}-1,3-fucosyltransferases VII. Our data suggest that in activated T cells that express both core 2 beta-1,6-N-acetylglucosaminyltransferase and {alpha}-1,3-fucosyltransferases VII (40), CD43 acts as one of the core proteins that are modified by these enzymes to carry selectin-binding fucosylated core 2 O-glycans. Thus, although naive T cells express CD43 with a likely antiadhesion function (23), T cell activation may switch the molecule to a proadhesive selectin ligand through the modification of its glycans. How these apparently opposing functions regulate T cell trafficking in vivo needs further investigation. Besides CD43, CD34 (41), podocalyxin (42), and endomucin (43) are other sialomucins reported to function as antiadhesive molecules that become proadhesive selectin ligands when appropriately modified in specific cell types.

Although PSGL-1 and CD43 both function as E-selectin ligands, DKO Th1 cells still migrated into the inflamed skin in an E-selectin-dependent manner, indicating the existence of at least one more E-selectin ligand. Our precipitation experiments using E-selectin-IgG brought down ESL-1, an E-selectin ligand that is expressed on mouse neutrophils (12), from DKO Th1 cells. It is thus possible that ESL-1 mediates the residual migration, although this has to be confirmed using a blocking anti-ESL-1 Ab or ESL-1-deficient cells. Note that whereas PSGL-1 and CD43 were both precipitated from activated human T cells with E-selectin-IgG (18), ESL-1 was not, even though it is expressed in human T cells (our unpublished data), suggesting that on human T cells ESL-1 may not function as an E-selectin ligand. In an earlier report, we observed that another band of ~180 kDa was precipitated with E-selectin-IgG from human T cells (18). This unidentified molecule may serve as another E-selectin ligand in human T cells.

PSGL-1 functions as an E-selectin ligand not only on activated T cells, but also on neutrophils (44). CD43 is abundant on neutrophils, raising the possibility that it functions as an E-selectin ligand on these cells as well. A previous study showed, however, that CD43–/– neutrophils exhibit not decreased, but rather enhanced interactions with immobilized E-selectin under flow conditions (45). In addition, PSGL-1, but not CD43, from the human promyelocytic cell line HL-60, carries selectin-binding fucosylated glycans (46). Our data also showed that in a CHS model, the CD43 deficiency did not detectably decrease the number of cells (predominantly neutrophils) infiltrating the dermis, even in the absence of PSGL-1. These observations may argue against a role for CD43 as a neutrophil E-selectin ligand. CD44 does serve as an E-selectin ligand on neutrophils (17), but our biochemical analyses did not detect CD44 as an E-selectin-binding protein on T cells, which abundantly express this molecule (18). It is possible that the repertoire of molecules serving as core proteins for E-selectin ligands is cell-type dependent, varying with the glycosyltransferase expression profile.

In conclusion, our study shows that CD43 serves as an E-selectin ligand on activated T cells and thereby mediates the migration of these cells to inflamed sites. To our knowledge, this is the first demonstration of an in vivo role of CD43 as an E-selectin ligand on T cells. The relative contributions of PSGL-1 and CD43 to E-selectin-mediated processes should be studied in various inflammatory and infectious conditions in vivo.


   Acknowledgments
 
We thank Dr. Bruce Furie for the PSGL-1-deficient mice and the anti- ESL-1 Ab, Dr. John Lowe for selectin-IgM constructs, and Dr. Barry Wolitzky for the 9A9 mAb.


   Disclosures
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 Disclosures
References
 
The authors have no financial conflict of interest.


   Footnotes
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 This work was supported by a grant-in-aid for the 21st Century Center of Excellence Program from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and a grant-in-aid for scientific research from the Japan Society for the Promotion of Science, Japan. Back

2 Address correspondence and reprint requests to Dr. Takako Hirata, The 21st Century Center of Excellence Program, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan. E-mail address: thirata{at}biken.osaka-u.ac.jp Back

3 Abbreviations used in this paper: LN, lymph node; CHS, contact hypersensitivity; ESL-1, E-selectin ligand-1; DKO, PSGL-1/CD43 double deficient; LCMV, lymphocytic choriomeningitis virus; PSGL-1, P-selectin glycoprotein ligand-1; SA, streptavidin; sLeX, sialyl LewisX; WT, wild type. Back

Received for publication September 21, 2006. Accepted for publication November 28, 2006.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 Disclosures
 References
 

  1. von Andrian, U. H., C. R. Mackay. 2000. T-cell function and migration: two sides of the same coin. N. Engl. J. Med. 343: 1020-1034. [Free Full Text]
  2. Miyasaka, M., T. Tanaka. 2004. Lymphocyte trafficking across high endothelial venules: dogmas and enigmas. Nat. Rev. Immunol. 4: 360-370. [Medline]
  3. Kupper, T. S., R. C. Fuhlbrigge. 2004. Immune surveillance in the skin: mechanisms and clinical consequences. Nat. Rev. Immunol. 4: 211-222. [Medline]
  4. Ley, K., G. S. Kansas. 2004. Selectins in T-cell recruitment to non-lymphoid tissues and sites of inflammation. Nat. Rev. Immunol. 4: 325-335. [Medline]
  5. Weninger, W., L. H. Ulfman, G. Cheng, N. Souchkova, E. J. Quackenbush, J. B. Lowe, U. H. von Andrian. 2000. Specialized contributions by {alpha}(1,3)-fucosyltransferase-IV and FucT-VII during leukocyte rolling in dermal microvessels. Immunity 12: 665-676. [Medline]
  6. Sunderkotter, C., K. Steinbrink, U. Henseleit, R. Bosse, A. Schwarz, D. Vestweber, C. Sorg. 1996. Activated T cells induce expression of E-selectin in vitro and in an antigen-dependent manner in vivo. Eur. J. Immunol. 26: 1571-1579. [Medline]
  7. Harari, O. A., J. F. McHale, D. Marshall, S. Ahmed, D. Brown, P. W. Askenase, D. O. Haskard. 1999. Endothelial cell E- and P-selectin up-regulation in murine contact sensitivity is prolonged by distinct mechanisms occurring in sequence. J. Immunol. 163: 6860-6866. [Abstract/Free Full Text]
  8. Jones, T. R., N. Shirasugi, A. B. Adams, T. C. Pearson, C. P. Larsen. 2003. Intravital microscopy identifies selectins that regulate T cell traffic into allografts. J. Clin. Invest. 112: 1714-1723. [Medline]
  9. Borges, E., W. Tietz, M. Steegmaier, T. Moll, R. Hallmann, A. Hamann, D. Vestweber. 1997. P-selectin glycoprotein ligand-1 (PSGL-1) on T helper 1 but not on T helper 2 cells binds to P-selectin and supports migration into inflamed skin. J. Exp. Med. 185: 573-578. [Abstract/Free Full Text]
  10. Hirata, T., G. Merrill-Skoloff, M. Aab, J. Yang, B. C. Furie, B. Furie. 2000. P-selectin glycoprotein ligand 1 (PSGL-1) is a physiological ligand for E-selectin in mediating T helper 1 lymphocyte migration. J. Exp. Med. 192: 1669-1676. [Abstract/Free Full Text]
  11. Lowe, J. B.. 2002. Glycosylation in the control of selectin counter-receptor structure and function. Immunol. Rev. 186: 19-36. [Medline]
  12. Steegmaier, M., A. Levinovitz, S. Isenmann, E. Borges, M. Lenter, H. P. Kocher, B. Kleuser, D. Vestweber. 1995. The E-selectin-ligand ESL-1 is a variant of a receptor for fibroblast growth factor. Nature 373: 615-620. [Medline]
  13. Picker, L. J., R. A. Warnock, A. R. Burns, C. M. Doerschuk, E. L. Berg, E. C. Butcher. 1991. The neutrophil selectin LECAM-1 presents carbohydrate ligands to the vascular selectins ELAM-1 and GMP-140. Cell 66: 921-933. [Medline]
  14. Zollner, O., M. C. Lenter, J. E. Blanks, E. Borges, M. Steegmaier, H. G. Zerwes, D. Vestweber. 1997. L-selectin from human, but not from mouse neutrophils binds directly to E-selectin. J. Cell Biol. 136: 707-716. [Abstract/Free Full Text]
  15. Jones, W. M., G. M. Watts, M. K. Robinson, D. Vestweber, M. A. Jutila. 1997. Comparison of E-selectin-binding glycoprotein ligands on human lymphocytes, neutrophils, and bovine {gamma}{delta} T cells. J. Immunol. 159: 3574-3583. [Abstract]
  16. Jutila, M. A., S. Kurk, L. Jackiw, R. N. Knibbs, L. M. Stoolman. 2002. L-selectin serves as an E-selectin ligand on cultured human T lymphoblasts. J. Immunol. 169: 1768-1773. [Abstract/Free Full Text]
  17. Katayama, Y., A. Hidalgo, J. Chang, A. Peired, P. S. Frenette. 2005. CD44 is a physiological E-selectin ligand on neutrophils. J. Exp. Med. 201: 1183-1189. [Abstract/Free Full Text]
  18. Matsumoto, M., K. Atarashi, E. Umemoto, Y. Furukawa, A. Shigeta, M. Miyasaka, T. Hirata. 2005. CD43 functions as a ligand for E-selectin on activated T cells. J. Immunol. 175: 8042-8050. [Abstract/Free Full Text]
  19. Fuhlbrigge, R. C., S. L. King, R. Sackstein, T. S. Kupper. 2006. CD43 is a ligand for E-selectin on CLA+ human T cells. Blood 107: 1421-1426. [Abstract/Free Full Text]
  20. Fukuda, M., S. Tsuboi. 1999. Mucin-type O-glycans and leukosialin. Biochim. Biophys. Acta 1455: 205-217. [Medline]
  21. Manjunath, N., R. S. Johnson, D. E. Staunton, R. Pasqualini, B. Ardman. 1993. Targeted disruption of CD43 gene enhances T lymphocyte adhesion. J. Immunol. 151: 1528-1534. [Abstract]
  22. Manjunath, N., M. Correa, M. Ardman, B. Ardman. 1995. Negative regulation of T-cell adhesion and activation by CD43. Nature 377: 535-538. [Medline]
  23. Stockton, B. M., G. Cheng, N. Manjunath, B. Ardman, U. H. von Andrian. 1998. Negative regulation of T cell homing by CD43. Immunity 8: 373-381. [Medline]
  24. McEvoy, L. M., H. Sun, J. G. Frelinger, E. C. Butcher. 1997. Anti-CD43 inhibition of T cell homing. J. Exp. Med. 185: 1493-1498. [Abstract/Free Full Text]
  25. Johnson, G. G., A. Mikulowska, E. C. Butcher, L. M. McEvoy, S. A. Michie. 1999. Anti-CD43 monoclonal antibody L11 blocks migration of T cells to inflamed pancreatic islets and prevents development of diabetes in nonobese diabetic mice. J. Immunol. 163: 5678-5685. [Abstract/Free Full Text]
  26. Onami, T. M., L. E. Harrington, M. A. Williams, M. Galvan, C. P. Larsen, T. C. Pearson, N. Manjunath, L. G. Baum, B. D. Pearce, R. Ahmed. 2002. Dynamic regulation of T cell immunity by CD43. J. Immunol. 168: 6022-6031. [Abstract/Free Full Text]
  27. Ford, M. L., T. M. Onami, A. I. Sperling, R. Ahmed, B. D. Evavold. 2003. CD43 modulates severity and onset of experimental autoimmune encephalomyelitis. J. Immunol. 171: 6527-6533. [Abstract/Free Full Text]
  28. Atarashi, K., T. Hirata, M. Matsumoto, N. Kanemitsu, M. Miyasaka. 2005. Rolling of Th1 cells via P-selectin glycoprotein ligand-1 stimulates LFA-1-mediated cell binding to ICAM-1. J. Immunol. 174: 1424-1432. [Abstract/Free Full Text]
  29. Austrup, F., D. Vestweber, E. Borges, M. Lohning, R. Brauer, U. Herz, H. Renz, R. Hallmann, A. Scheffold, A. Radbruch, A. Hamann. 1997. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflamed tissues. Nature 385: 81-83. [Medline]
  30. Tietz, W., Y. Allemand, E. Borges, D. von Laer, R. Hallmann, D. Vestweber, A. Hamann. 1998. CD4+ T cells migrate into inflamed skin only if they express ligands for E- and P-selectin. J. Immunol. 161: 963-970. [Abstract/Free Full Text]
  31. Wang, B., H. Fujisawa, L. Zhuang, I. Freed, B. G. Howell, S. Shahid, G. M. Shivji, T. W. Mak, D. N. Sauder. 2000. CD4+ Th1 and CD8+ type 1 cytotoxic T cells both play a crucial role in the full development of contact hypersensitivity. J. Immunol. 165: 6783-6790. [Abstract/Free Full Text]
  32. Hirata, T., B. C. Furie, B. Furie. 2002. P-, E-, and L-selectin mediate migration of activated CD8 + T lymphocytes into inflamed skin. J. Immunol. 169: 4307-4313. [Abstract/Free Full Text]
  33. Xu, H., N. A. DiIulio, R. L. Fairchild. 1996. T cell populations primed by hapten sensitization in contact sensitivity are distinguished by polarized patterns of cytokine production: interferon {gamma}-producing (Tc1) effector CD8+ T cells and interleukin (IL) 4/IL-10-producing (Th2) negative regulatory CD4+ T cells. J. Exp. Med. 183: 1001-1012. [Abstract/Free Full Text]
  34. Gorbachev, A. V., R. L. Fairchild. 2004. CD4+ T cells regulate CD8+ T cell-mediated cutaneous immune responses by restricting effector T cell development through a Fas ligand-dependent mechanism. J. Immunol. 172: 2286-2295. [Abstract/Free Full Text]
  35. Ostberg, J. R., R. K. Barth, J. G. Frelinger. 1998. The Roman god Janus: a paradigm for the function of CD43. Immunol. Today 19: 546-550. [Medline]
  36. Rosenstein, Y., J. K. Park, W. C. Hahn, F. S. Rosen, B. E. Bierer, S. J. Burakoff. 1991. CD43, a molecule defective in Wiskott-Aldrich syndrome, binds ICAM-1. Nature 354: 233-235. [Medline]
  37. Baum, L. G., M. Pang, N. L. Perillo, T. Wu, A. Delegeane, C. H. Uittenbogaart, M. Fukuda, J. J. Seilhamer. 1995. Human thymic epithelial cells express an endogenous lectin, galectin-1, which binds to core 2 O-glycans on thymocytes and T lymphoblastoid cells. J. Exp. Med. 181: 877-887. [Abstract/Free Full Text]
  38. Van den Berg, T. K., D. Nath, H. J. Ziltener, D. Vestweber, M. Fukuda, I. van Die, P. R. Crocker. 2001. Cutting edge: CD43 functions as a T cell counterreceptor for the macrophage adhesion receptor sialoadhesin (Siglec-1). J. Immunol. 166: 3637-3640. [Abstract/Free Full Text]
  39. Piccio, L., B. Rossi, L. Colantonio, R. Grenningloh, A. Gho, L. Ottoboni, J. W. Homeister, E. Scarpini, M. Martinello, C. Laudanna, et al 2005. Efficient recruitment of lymphocytes in inflamed brain venules requires expression of cutaneous lymphocyte antigen and fucosyltransferase-VII. J. Immunol. 174: 5805-5813. [Abstract/Free Full Text]
  40. Wagers, A. J., C. M. Waters, L. M. Stoolman, G. S. Kansas. 1998. Interleukin 12 and interleukin 4 control T cell adhesion to endothelial selectins through opposite effects on {alpha}1,3-fucosyltransferase VII gene expression. J. Exp. Med. 188: 2225-2231. [Abstract/Free Full Text]
  41. Drew, E., J. S. Merzaban, W. Seo, H. J. Ziltener, K. M. McNagny. 2005. CD34 and CD43 inhibit mast cell adhesion and are required for optimal mast cell reconstitution. Immunity 22: 43-57. [Medline]
  42. Takeda, T., W. Y. Go, R. A. Orlando, M. G. Farquhar. 2000. Expression of podocalyxin inhibits cell-cell adhesion and modifies junctional properties in Madin-Darby canine kidney cells. Mol. Biol. Cell 11: 3219-3232. [Abstract/Free Full Text]
  43. Kinoshita, M., T. Nakamura, M. Ihara, T. Haraguchi, Y. Hiraoka, K. Tashiro, M. Noda. 2001. Identification of human endomucin-1 and -2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity. FEBS Lett. 499: 121-126. [Medline]
  44. Xia, L., M. Sperandio, T. Yago, J. M. McDaniel, R. D. Cummings, S. Pearson-White, K. Ley, R. P. McEver. 2002. P-selectin glycoprotein ligand-1-deficient mice have impaired leukocyte tethering to E-selectin under flow. J. Clin. Invest. 109: 939-950. [Medline]
  45. Woodman, R. C., B. Johnston, M. J. Hickey, D. Teoh, P. Reinhardt, B. Y. Poon, P. Kubes. 1998. The functional paradox of CD43 in leukocyte recruitment: a study using CD43-deficient mice. J. Exp. Med. 188: 2181-2186. [Abstract/Free Full Text]
  46. Wilkins, P. P., R. P. McEver, R. D. Cummings. 1996. Structures of the O-glycans on P-selectin glycoprotein ligand-1 from HL-60 cells. J. Biol. Chem. 271: 18732-18742. [Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 J. Immunol.Home page
M. Matsumoto, A. Shigeta, M. Miyasaka, and T. Hirata
CD43 Plays Both Antiadhesive and Proadhesive Roles in Neutrophil Rolling in a Context-Dependent Manner
J. Immunol., September 1, 2008; 181(5): 3628 - 3635.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
S. C. Kerr, C. B. Fieger, K. R. Snapp, and S. D. Rosen
Endoglycan, a Member of the CD34 Family of Sialomucins, Is a Ligand for the Vascular Selectins
J. Immunol., July 15, 2008; 181(2): 1480 - 1490.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
M. E. Gainers, L. Descheny, S. R. Barthel, L. Liu, M.-A. Wurbel, and C. J. Dimitroff
Skin-Homing Receptors on Effector Leukocytes Are Differentially Sensitive to Glyco-Metabolic Antagonism in Allergic Contact Dermatitis
J. Immunol., December 15, 2007; 179(12): 8509 - 8518.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Matsumoto, M.
Right arrow Articles by Hirata, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Matsumoto, M.
Right arrow Articles by Hirata, T.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS