| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Research Laboratories, Yoshitomi Pharmaceutical Industries, Limited,Iruma, Saitama, Japan
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We previously reported that a potent immunosuppressive compound, ISP-I, and its derivatives, mycestericins, were isolated from the culture broth of Isaria sinclairii, a species of vegetative wasp (9, 10). Chemical modification of ISP-I led to a novel synthetic immunosuppressant, 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol hydrochloride (FTY720), which has more potent immunosuppressive activity and less toxicity than ISP-I (11, 12, 13, 14). FTY720, at 0.1 mg/kg or more, significantly prolonged skin or cardiac allograft survival and host survival in lethal graft-vs-host reaction in rats (15, 16, 17). In addition, combination treatment with FTY720 and a subtherapeutic dose of CsA resulted in a synergistic effect on canine renal allografts as well as rat skin or cardiac allografts (15, 16, 18, 19). A striking feature of FTY720 is induction of a marked decrease in the number of PBL, especially T cells, at doses that prolong allograft survival (15, 16). It has been hypothesized that the decrease in lymphocyte number is caused by apoptotic cell death of lymphocytes, because FTY720 at 4 µM (1.4 µg/ml) or more induces apoptosis of rat spleen cells and human peripheral blood cells in vitro (20, 21). However, the trough level of the blood concentration of FTY720 in dogs given 5 mg/kg is less than 200 ng/ml (21). In addition, the blood concentration range of FTY720 is 0.2 to 20 ng/ml when given to rats at 0.1 mg/kg to 1 mg/kg (unpublished data from our laboratories). Thus, the hypothesis concerning FTY720-induced apoptosis is insufficient to explain the intrinsic mechanism of the decreasing effect on PBL number by FTY720, because it is clearly impossible for FTY720 to induce apoptotic cell death of lymphocytes at a dose range of 0.1 to 1 mg/kg in vivo.
The immunologically mature lymphocytes are known to continuously recirculate in the peripheral blood, spleen, lymphatic vessels, TDL, PLN, MLN, and PP (22). It has also been reported that lymphocyte recirculation is regulated by lymphocyte trafficking via lymphocyte-homing receptors, including CD62L, CD49d/ß7 integrin, and CD11a/CD18 (22, 23, 24, 25, 26). We focused on lymphocyte recirculation in vivo and hypothesized that, if lymphocyte homing is modulated by FTY720, the number of PBL would decrease without death of lymphocytes. In this paper, we will describe that the marked decrease in the number of PBL induced by FTY720 is due to acceleration of lymphocyte homing to PLN, MLN, and PP via lymphocyte-homing receptors.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Inbred male F344 rats (RT1lv1) and WKAH rats (RT1k) were purchased from Japan Charles River (Atsugi, Kanagawa, Japan) and Japan SLC (Hamamatsu, Shizuoka, Japan), respectively. All rats were used at 4 to 12 wk of age.
Cell lines
A mouse IL-2-dependent cytotoxic T cell line, CTLL-2 (27), was obtained from the American Type Culture Collection (ATCC, Rockville, MD) and maintained in RPMI 1640 medium supplemented with 2 mM L-glutamine, 10 mM HEPES, 100 U/ml penicillin, 60 µg/ml kanamycin sulfonate, 50 µM 2-ME, 10% FCS (Boehringer Mannheim, Mannheim, Germany) and 20% Con A-stimulated rat spleen cell culture supernatant prepared by the method previously described (28).
Reagents
FTY720 was synthesized according to the method previously
described (13). The chemical structure of FTY720 is shown in Figure 1
. For i.v. injection, FTY720 was
dissolved in 1% 
-cyclodextrin and 5% mannitol solution. CsA
(Sandimmun, for i.v. injection; Sandoz, Basel, Switzerland) and FK506
(Prograf, for i.v. injection, Fujisawa Pharmaceutical, Osaka, Japan)
was diluted with saline. For oral administration, FTY720 was dissolved
in distilled water. CsA (Sandimmun, for oral solution) and FK506
(Prograf, for i.v. injection) were diluted with olive oil (Sigma
Chemicals, St. Louis, MO) and with distilled water, respectively.
Control animals received the vehicle only. For in vitro treatments,
FTY720, CsA (Sandimmun, for i.v. injection) and FK506 (Prograf, for
i.v. injection) were dissolved in saline and diluted to the appropriate
concentrations with RPMI 1640 medium containing 10% FCS.
|
FITC-conjugated anti-rat CD3 mAb (1F4; 29 was obtained from Caltag Laboratories (South San Francisco, CA). Biotinylated anti-rat CD3 mAb (G4.18; 30 and phycoerythrin (PE)-conjugated anti-rat CD45RA or A/B mAb (OX-33; 31 , FITC-conjugated anti-rat CD4 mAb (OX-38; 32 , PE-conjugated anti rat CD8 mAb (OX-8; 33 and streptavidin-Cy-chrome conjugate were obtained from PharMingen (La Jolla, CA). Hamster anti-rat CD62L mAb (HRL3; 34 , mouse anti-rat CD49d mAb (TA-2; 35 , mouse anti-rat CD11a mAb (clone WT.1; 36 , and biotinylated TA-2 were purchased from Seikagaku-kogyo (Tokyo, Japan). PE-conjugated HRL-3, PE-conjugated WT.1, and isotype-matched control IgGs were obtained from PharMingen.
Rat skin allograft
MHC-incompatible rat skin allograft was performed by the method described previously, with WKAH rats (RT1k) as donors and F344 rats (RT1lv1) as recipients (15). Briefly, full-thickness skin grafts (2.0 x 2.0 cm2 pieces) from donor rats were transplanted to the lateral thorax of the recipient rats and covered with sterile bactericidal gauze. The entire chest was then wrapped with an elastic bandage. The dressings were removed on day 5 and the grafts were inspected daily until rejection, which was defined as more than 90% necrosis of the graft epithelium. FTY720, CsA, or FK506 was administered daily to the allografted recipients for 14 days from the day of transplantation.
IL-2 production in allogeneic MLC in rats
Effects on IL-2 production in rat allogeneic MLC were evaluated according to the method previously reported (28). Allogeneic MLC was performed by using nylon nonadherent splenic lymphocytes from F344 rats as responder cells and mitomycin C (Kyowa Hakko, Tokyo, Japan)-pretreated spleen cells from WKAH rats as stimulator cells. Responder cells at 5 x 106 cells/well were cocultured with an equal number of stimulator cells in 2.0 ml of RPMI 1640 medium containing 50 µM 2-ME and 10% FCS. After culturing for 48 h at 37°C in 5% CO2, the culture supernatants were collected, and their IL-2 activities were determined by CTLL-2 proliferation assay (27). Briefly, CTLL-2 cells (104 cells/well) were cultured in the presence of serial twofold dilution of culture supernatants for 20 h at 37°C, pulsed with 0.5 µCi of [3H]thymidine (TdR) (Amersham, Tokyo, Japan) for 4 h at 37°C in 5% CO2 and then harvested onto glass fiber filters using an automatic cell harvester. The radioactivity incorporated into the cells was determined by a scintillation counter (1450 MicroBeta; Pharmacia Biotech, Uppsala, Sweden). IL-2 activity in the supernatants is expressed as U/ml in comparison with recombinant rat IL-2 (Genzyme, Cambridge MA) as a standard (27).
Flow cytometry
Peripheral blood was collected from the tail veins of F344 rats. Spleen, PLN (axillary lymph nodes were used as PLN in this study), MLN, and PP were removed from rats, and single cell suspensions were prepared by mincing and passing through stainless mesh. Lymphocytes in TDL were collected by cannulation of thoracic duct under anesthesia according to the method described previously (37). Flow cytometry analysis was performed by using EPICS XL-MCL (Coulter, Miami, FL). Lymphocytes from rat peripheral blood, TDL, spleen, PLN, MLN, or PP were stained with FITC-anti-rat CD3 mAb (1F4) and PE-anti-rat CD45RA or A/B mAb (OX-33), which is reported to bind B cells only (31). The number of total lymphocytes was determined by the lymphocyte gating method. The numbers of T cells and B cells were determined by two-color flow cytometry. The proportions of CD4+ T cells and CD8+ T cells were determined by three-color flow cytometry using biotinylated-anti-rat CD3 mAb (G4.18), FITC-anti-rat CD4 (OX-38), PE-anti-rat CD8 (OX-8), and streptavidin-Cy-chrome conjugate. Expression of lymphocyte-homing receptors on rat T cells was determined by two-color flow cytometry using FITC-anti-rat CD3 (IF4), PE-anti-rat CD62L mAb (HRL3), biotinylated-anti-rat CD49d mAb (TA-2), PE-anti-rat CD11a mAb (WT.1), and streptavidin-Cy-Chrome conjugate.
Analysis of lymphocyte homing with calcein-labeled lymphocytes
Lymphocytes (1 x 108 cells) from PLN and MLN of F344 rats were labeled by incubation for 30 min on ice in 10 ml of RPMI 1640 medium containing 0.2 µM calcein-AM (Molecular Probes, Eugene, OR) (38). After labeling with calcein, the viability of lymphocytes was more than 94% by trypan blue dye exclusion test. The calcein-labeled lymphocytes (5 x 107 cells/0.5 ml) were i.v. transfused at 2.5 h after FTY720 administration to F344 rats. Then, the rats were sacrificed 30 min after the transfusion, and the peripheral blood, spleen, PLN, MLN, and PP were collected. The number of calcein-labeled lymphocytes in these tissues was determined by flow cytometry. To examine the effect of mAbs against lymphocyte-homing receptors, the calcein-labeled lymphocytes were treated with 60 µg/ml of hamster anti-rat CD62L mAb (HRL3), mouse anti-rat CD49d mAb (TA-2), mouse anti-rat CD11a mAb (WT.1), or all three mAbs at 4°C for 30 min. As a control, isotype-matched irrelevant IgGs were used under the same conditions. After the treatment of mAbs, the calcein-labeled lymphocytes were transfused i.v. to the rats, and their tissue distribution was determined as described above.
Statistical analysis
The statistical differences in allograft survival time compared with vehicle-treated control group were calculated by the generalized Wilcoxon test with Hommel’s multiple comparison test. In other experiments, statistical differences compared with the vehicle-treated control were calculated by Dunnett’s test. Differences between groups were considered significant at p < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
To clarify the efficacy and potency of the immunosuppressive
activity of FTY720, the prolonging effect of FTY720, CsA, and FK506 on
rat skin allograft survival was examined in MHC-incompatible rat
strains of WKAH donors and F344 recipients. The immunosuppressants were
administered i.v. or orally for 14 days from the day of the
transplantation. In this skin allograft models, all grafts in the
control (vehicle-treated) group were rejected 6 to 7 days after
transplantation. As shown in Figure 2,
FTY720 at 0.03 mg/kg or higher doses significantly prolonged allograft
survival in a dose-dependent manner by either i.v. or oral
administration. CsA and FK506 were also effective at doses of 3 mg/kg
or more and 0.3 mg/kg or more, respectively, in this model. These
results indicate that FTY720 possesses more potent immunosuppressive
activity than CsA or FK506 on allograft rejection in an
MHC-incompatible combination.
|
The effect of FTY720 on IL-2 production in allogeneic MLC in rats
was examined in comparison with that of CsA and FK506. The results are
shown in Figure 3. CsA and FK506
dose-dependently inhibited IL-2 production in rat allogeneic MLC,
consistent with the results of previous studies in mice and humans
(3, 4, 5). The IC50 values (concentrations that inhibit
50%) of CsA and FK506 for IL-2 production were 3.5 nM and 0.043 nM,
respectively. By contrast, FTY720 up to 1000 nM did not affect IL-2
production in rat allogeneic MLC (Fig. 3) and failed to inhibit T cell
proliferation by alloantigen stimulation or IL-2-dependent
proliferation of CTLL-2 cells (data not shown). These findings suggest
that FTY720 exerts a potent immunosuppressive effect by a mechanism
distinct from that of CsA and FK506.
|
As described in our previous papers, FTY720 significantly
decreases the number of PBLs in allografted rats, especially the number
of T cells (15, 16). To clarify the mechanism of the decrease in
the number of lymphocytes by FTY720, the tissue distribution of
lymphocytes was analyzed in the peripheral blood, TDL, spleen, PLN,
MLN, and PP of F344 rats, following a single oral administration of
FTY720 (0.1 and 1 mg/kg). The proportions of lymphocytes, T cells, T
cell subsets (CD4+ or CD8+ T cells), and B
cells were determined by flow cytometry. Figure 4 shows the time course of lymphocyte, T
cell, and B cell numbers in blood, TDL, and spleen after a single oral
administration of FTY720. The numbers of total lymphocytes, T cells,
and B cells in blood and TDL dramatically decreased to less than 10%
of the control values within 3 to 24 h after administration. The
decrease in the number of lymphocytes by FTY720 was more marked in TDL
than in peripheral blood. The numbers of splenic lymphocytes, T cells,
and B cells were also significantly decreased by FTY720 treatment to
40% to 80% of the control with a time course similar to that of the
PBLs. Lymphocyte numbers in blood and spleen had recovered to the
control level on day 7 after administration. Although the number of
lymphocytes in TDL was still decreased to 20% to 40% of the control
by FTY720 administration, there was almost complete recovery within 2
wk (data not shown). By contrast, the numbers of lymphocytes in PLN,
MLN, and PP were significantly increased in a dose-dependent manner
after administration of FTY720 (Fig. 5).
The number of PLN lymphocytes reached a maximum at 12 h after
FTY720 administration and returned to the control level by 24 h.
Lymphocytes in MLN and PP also increased to 180% and 300%,
respectively, of the controls at 24 h after FTY720 administration
and then returned to the control level within 5 days. The time courses
of the numbers of T cells and B cells in PLN, MLN, and PP were similar
to that of the total lymphocyte number. The increase in numbers of T
cells was especially marked in PLN, MLN, and PP. These findings
indicate that FTY720 modulates the tissue distribution of lymphocytes
in blood, TDL, spleen, PLN, MLN, and PP in rats. Figure 6, A and B, shows
the proportions of T cells, B cells, or T cell subsets
(CD4+ or CD8+ T cells) in these lymphoid
tissues 12 h after administration of FTY720. The numbers of T
cells, B cells, CD4+ T cells or CD8+ T cells
showed changes similar to the total numbers of lymphocytes in all
tissues tested. By contrast, FTY720 did not cause any clear changes in
the number of red blood cells, thymocytes (CD4+,
CD8+ or CD4+CD8+ subpopulation), or
bone marrow cells 12 h after administration of FTY720 (Fig. 6C). Thus, the changes in lymphocyte distribution
induced by FTY720 appear to be specific for mature lymphocytes but do
lack selectivity for T cells, B cells, or T cell subsets.
|
|
|
To determine the effect of FTY720 on lymphocyte trafficking
between blood to various lymphoid tissues in rats, calcein-labeled
lymphocytes were i.v. transfused to the strain- and sex-matched F344
rats 2.5 h after administration of FTY720. The rats were
sacrificed 30 min later, and the tissue distribution of calcein-labeled
lymphocytes was determined by flow cytometry. Figure 7 shows the number of calcein-labeled
lymphocytes found in the PLN, MLN, PP, spleen, and blood of rats given
FTY720 (0.1 and 1 mg/kg) orally as compared with vehicle-treated
control rats. FTY720 significantly increased the number of
calcein-labeled lymphocytes in PLN, MLN, and PP in a dose-dependent
manner but decreased in spleen and blood. These results indicate that
FTY720 accelerates lymphocyte homing from blood or spleen to PLN, MLN,
and PP. In addition, the involvement of lymphocyte-homing receptors in
FTY720-induced acceleration of lymphocyte homing was assessed by
pretreatment with mAb against CD62L, CD49d, or CD11a with
calcein-labeled lymphocytes. With a similarity to the results in
previous studies (39, 40), anti-CD62L mAb and CD49d mAb prevented
normal lymphocyte homing to PLN and MLN, whereas CD11a mAb partially
inhibited homing to PLN, MLN, and PP (Fig. 8A). Anti-CD62 mAb
inhibited normal lymphocyte homing by 90% in PLN, 70% in MLN, and
50% in PP. Treatment with CD49d mAb, on the other hand, resulted in
inhibition of normal lymphocyte homing by 20% in PLN, 60% in MLN, and
80% in PP. Treatment with anti-CD11a mAb displayed a partial
inhibition of normal lymphocyte homing by 35% to 60% in these
lymphoid tissues. Simultaneous treatment with these mAbs resulted in
marked inhibition (>90%) of normal lymphocyte homing in these
lymphoid tissues. As shown in Figure 8B, FTY720-induced
lymphocyte homing, as well as normal lymphocyte homing, was prevented
by treatment with anti-CD49d or CD11a mAb. Unlike normal lymphocyte
homing, CD62L mAb inhibited the FTY720-induced lymphocyte homing by
85% in PP, 70% in MLN, and 50% in PLN. In addition, FTY720-induced
lymphocyte homing is almost completely blocked (>90% inhibition) by
simultaneous treatment with mAbs against CD49d, CD62L, and CD11a in
PLN, MLN, and PP. These results suggest that FTY720-induced
acceleration of lymphocyte homing, as well as normal lymphocyte homing,
is mediated by lymphocyte-homing receptors, including CD62L,
CD49d/ß7 integrin, and CD11a/CD18.
|
|
Since the acceleration of lymphocyte homing by FTY720 appears to
be mediated by lymphocyte-homing receptors, there is a possibility that
FTY720 up-regulates the expression of lymphocyte-homing receptors on
lymphocytes. To determine whether FTY720 induces up-regulation, the
expression of CD62L, CD49d, and CD11a on T cells was analyzed in rats
at 1 h in blood and at 3 h in MLN after single oral
administration of FTY720. The expression of these receptors could not
be determined in blood at 3 h after administration, because of the
disappearance of T cells from the peripheral blood. Figure 9 shows the typical CD62L, CD49d, and
CD11a levels on T cells of rats given FTY720 at 1 mg/kg orally. FTY720
did not have any up-regulating effect on CD62L, CD49d, or CD11a
expression on T cells in blood or MLN.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
). CsA and FK506 are
known to exert immunosuppressive activity by inhibiting the production
of Th1 cell-associated cytokines in Ag-stimulated helper T cells (3, 4). As reported previously, FTY720, unlike CsA, did not inhibit IL-2
production by Con A-stimulated T cells in rats (15). Consistent with
the results of mitogen-stimulated IL-2 production, FTY720 did not
affect IL-2 production by alloantigen stimulation in rats (Fig. 3).
Since FTY720 did not affect the process of T cell activation, including
IL-2 production or IL-2-dependent T cell proliferation, FTY720
presumably possesses a unique immunosuppressive mechanism of action
distinct from that of CsA and FK506 and thus shows a synergistic effect
on allograft survival when combined with CsA (15, 16).
The most striking feature of FTY720 is the induction of a
dramatic decrease in number of lymphocytes in peripheral blood and TDL.
Oral administration of FTY720 at 0.1 mg/kg or more selectively
decreased the number of lymphocytes to extremely low levels in blood
and TDL within 3 to 24 h after administration. Similar lymphopenia
is known to be induced by treatment with steroids or cyclophosphamide
(41, 42). However, steroids and cyclophosphamide markedly decrease the
number of PBLs and immunologically incompetent thymocytes associating
atrophy of thymic cortex. By contrast, FTY720 did not have any clear
effect on the numbers of thymocytes and bone marrow cells in rats (Fig. 6C). We also confirmed that FTY720 did not affect the
corticosteriod levels in peripheral blood (our unpublished data). From
these findings, we presume that the decrease in the number of PBLs by
FTY720 is not due to inhibition of intrathymic differentiation of T
cells or corticosteriod induction in vivo. Thus, the decrease in the
number of lymphocytes by FTY720 is likely to be selective for
immunologically mature lymphocytes, which have a capability of
lymphocyte trafficking between blood and lymphoid tissues, recognizing
foreign antigens, and inducing both cell-mediated and humoral immune
responses.
The time course studies of lymphocyte number in blood and
lymphoid tissues revealed that lymphocytes decreased in blood, TDL, and
spleen but increased in PLN, MLN, and PP within 3 to 24 h after
FTY720 administration to rats (Figs. 4 and 5). In addition, the results
of lymphocyte-trafficking studies by transfusion of calcein-labeled
lymphocytes confirmed that the trafficking of circulating lymphocytes
to PLN, MLN, and PP in rats was accelerated at 3 h after FTY720
administration (Fig. 7). From these findings, we conclude that FTY720
sequesters circulating mature lymphocytes into PLN, MLN, and PP by
acceleration of lymphocyte homing and thereby decreases the number of
lymphocytes in blood, TDL, and spleen.
Lymphocyte trafficking between blood and lymphoid tissues,
including PLN, MLN, and PP is known to be regulated and dependent on
the expression of specific cell surface adhesion molecules (22, 23, 24, 25, 26, 39, 40). Recirculation of lymphocytes consists of trafficking from blood to
PLN, MLN, and PP and returning to peripheral blood via lymphatic
vessels and TDL. The homing of circulating lymphocytes to PLN, MLN, and
PP was reported to be mediated by the attachment of lymphocytes to HEV
in these lymphoid tissues (22, 23, 24, 25, 26). The attachment of lymphocytes
to HEV is involved in adhesion between lymphocyte-homing receptors,
including CD62L (L-selectin), CD49d/ß7 integrin
(lymphocyte PP adhesion molecule-1 (LPAM-1)), and CD11a/CD18 (LFA-1),
and their ligands mucosal addressin cell adhesion molecule-1
(MAdCAM-1), glycosylation-dependent cell adhesion molecule-1
(GlyCAM-1), and ICAM-1 (22, 23, 24, 25, 26, 39, 40). In the present study,
FTY720-induced lymphocyte homing to PLN, MLN, and PP was completely
blocked by simultaneous treatment with mAbs against CD49d, CD62L, and
CD11a (Fig. 8). These findings indicate that FTY720 accelerates
lymphocyte homing mediated by lymphocyte-homing receptors, including
CD62L, CD49d/ß7 integrin, and CD11a/CD18. However,
anti-CD62L-mAb treatment resulted in different patterns between
normal and FTY720-induced lymphocyte homings to PLN, MLN, and PP in
rats. Normal lymphocyte homing to PP is predominantly mediated by
CD49d/ß7 integrin, and partially by CD62L. In contrast,
involvement of CD62L appeared to be more dominant in FTY720-induced
lymphocyte homing than in normal lymphocyte homing to PP. In other
experiments, expression of CD62L, CD49d, and CD11a was unaffected by
FTY720. Based on these results, there is a possibility that FTY720
promotes adhesion between lymphocytes and HEV by enhancing
avidity/affinity between adhesion molecules and ligands, including
activation of integrins. Since GlyCAM-1 (43) and macrophage
inflammatory protein-1 (MIP-1ß) (44) have been reported to be
triggering molecules for integrin activation, FTY720 may promote
adhesion between lymphocytes and HEV by induction of these triggering
molecules. We are currently investigating the effect of FTY720 on
activation of integrins and expression of ligands for lymphocyte-homing
receptors. We are also studying the direct effect of FTY720 on
lymphocyte-HEV interaction, by using an established rat HEV cell line
and the Stamper-Woodruff assay (45) in vitro.
From the results of the analysis of lymphocyte distribution in rats, it is likely that FTY720-induced reduction of mature-lymphocytes in peripheral blood, TDL, and spleen is predominantly caused by sequestration of lymphocytes in PLN, MLN, and PP, and not by cytotoxicity. Lymphocyte trafficking appears to play an important role in the initiation of both cell-mediated and humoral immune responses to foreign antigens (22, 39, 40). Recently, CD62L (L-selectin)-deficient mice were established and analyzed for lymphocyte distribution (46, 47). In contrast to the altered distribution of lymphocytes in FTY720-treated rats, L-selectin-deficient mice were reported to show 70% to 90% reduction in the number of PLN lymphocytes and 30% to 55% increase in spleen cellularity. L-selectin-deficient mice also display impaired delayed-type hypersensitivity and contact hypersensitivity response due to decreased recruitment of leukocytes into sites of inflammation, but immune response in spleen was augmented (47). From these results, it is suggested that systemic immune responses depend on lymphocyte recruitment and localization. Accordingly, if most of the circulating mature lymphocytes rapidly are sequestered in PLN, MLN, and PP, the systemic immune responses would be markedly suppressed. Based on these aspects, the sequestration of circulating mature lymphocytes is presumably a main mechanism of the immunosuppressive activity of FTY720. In our succeeding paper, we will report that FTY720-induced sequestration of circulating mature lymphocytes causes inhibition of intragraft immune responses, including T cell recruitment, and that FTY720 shows a striking synergistic effect when combined with CsA in a rat allograft model.
|
Footnotes |
|---|
2 Abbreviations used in this paper: CsA, cyclosporin A; FK506, tacrolimus; TDL, thoracic duct lymph; PLN, peripheral lymph nodes; MLN, mesenteric lymph nodes; PP, Peyer’s patches; HEV, high endothelial venules; FTY720, 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol hydrochloride; p.o., by mouth; PE, phycoerythrin.
Received for publication October 17, 1997. Accepted for publication January 12, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
4 integrins in lymphocyte homing to mucosal tissue in vivo. J. Immunol. 152:3282.[Abstract]
4ß7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell 74:185.[Medline]
This article has been cited by other articles:
|
|
 |
|
  G. Cattoretti, J. Mandelbaum, N. Lee, A. H. Chaves, A. M. Mahler, A. Chadburn, R. Dalla-Favera, L. Pasqualucci, and A. J. MacLennan Targeted Disruption of the S1P2 Sphingosine 1-Phosphate Receptor Gene Leads to Diffuse Large B-Cell Lymphoma Formation Cancer Res., November 15, 2009; 69(22): 8686 - 8692. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Gao, B. Majchrzak-Kita, E. N. Fish, and J. L. Gommerman Dynamic accumulation of plasmacytoid dendritic cells in lymph nodes is regulated by interferon-{beta} Blood, September 24, 2009; 114(13): 2623 - 2631. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. J. E. Raveney, D. A. Copland, L. B. Nicholson, and A. D. Dick Fingolimod (FTY720) as an Acute Rescue Therapy for Intraocular Inflammatory Disease Arch Ophthalmol, October 1, 2008; 126(10): 1390 - 1395. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Weinreich and K. A. Hogquist Thymic Emigration: When and How T Cells Leave Home J. Immunol., August 15, 2008; 181(4): 2265 - 2270. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Tomura, N. Yoshida, J. Tanaka, S. Karasawa, Y. Miwa, A. Miyawaki, and O. Kanagawa Monitoring cellular movement in vivo with photoconvertible fluorescence protein "Kaede" transgenic mice PNAS, August 5, 2008; 105(31): 10871 - 10876. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Metzler, P. Gfeller, G. Wieczorek, J. Li, B. Nuesslein-Hildesheim, A. Katopodis, M. Mueller, and V. Brinkmann Modulation of T cell homeostasis and alloreactivity under continuous FTY720 exposure Int. Immunol., May 1, 2008; 20(5): 633 - 644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Saab, A. Almony, K. J. Blinder, R. Schuessler, and D. C. Brennan Reversible Cystoid Macular Edema Secondary to Fingolimod in a Renal Transplant Recipient Arch Ophthalmol, January 1, 2008; 126(1): 140 - 141. [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Allende, D. Zhou, D. N. Kalkofen, S. Benhamed, G. Tuymetova, C. Borowski, A. Bendelac, and R. L. Proia S1P1 receptor expression regulates emergence of NKT cells in peripheral tissues FASEB J, January 1, 2008; 22(1): 307 - 315. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kim, M. Kim, N. Kim, V. D. D'Agati, C. W. Emala Sr, and H. T. Lee Isoflurane mediates protection from renal ischemia-reperfusion injury via sphingosine kinase and sphingosine-1-phosphate-dependent pathways Am J Physiol Renal Physiol, December 1, 2007; 293(6): F1827 - F1835. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. P. Coelho, S. G. Payne, R. Bittman, S. Spiegel, and C. Sato-Bigbee The Immunomodulator FTY720 Has a Direct Cytoprotective Effect in Oligodendrocyte Progenitors J. Pharmacol. Exp. Ther., November 1, 2007; 323(2): 626 - 635. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Kunisawa, Y. Kurashima, M. Higuchi, M. Gohda, I. Ishikawa, I. Ogahara, N. Kim, M. Shimizu, and H. Kiyono Sphingosine 1-phosphate dependence in the regulation of lymphocyte trafficking to the gut epithelium J. Exp. Med., October 1, 2007; 204(10): 2335 - 2348. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H. Huang, L. I. Cardenas-Navia, C. C. Caldwell, T. J. Plumb, C. G. Radu, P. N. Rocha, T. Wilder, J. S. Bromberg, B. N. Cronstein, M. Sitkovsky, et al. Requirements for T Lymphocyte Migration in Explanted Lymph Nodes J. Immunol., June 15, 2007; 178(12): 7747 - 7755. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kunisawa, Y. Kurashima, M. Gohda, M. Higuchi, I. Ishikawa, F. Miura, I. Ogahara, and H. Kiyono Sphingosine 1-phosphate regulates peritoneal B-cell trafficking for subsequent intestinal IgA production Blood, May 1, 2007; 109(9): 3749 - 3756. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Maeda, H. Matsuyuki, K. Shimano, H. Kataoka, K. Sugahara, and K. Chiba Migration of CD4 T Cells and Dendritic Cells toward Sphingosine 1-Phosphate (S1P) Is Mediated by Different Receptor Subtypes: S1P Regulates the Functions of Murine Mature Dendritic Cells via S1P Receptor Type 3 J. Immunol., March 15, 2007; 178(6): 3437 - 3446. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Deng, J. A. Clemas, G. Chrebet, P. Fischer, J. J. Hale, Z. Li, S. G. Mills, J. Bergstrom, S. Mandala, R. Mosley, et al. Identification of Leu276 of the S1P1 Receptor and Phe263 of the S1P3 Receptor in Interaction with Receptor Specific Agonists by Molecular Modeling, Site-Directed Mutagenesis, and Affinity Studies Mol. Pharmacol., March 1, 2007; 71(3): 724 - 735. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Daniel, N. Sartory, N. Zahn, G. Geisslinger, H. H. Radeke, and J. M. Stein FTY720 Ameliorates Th1-Mediated Colitis in Mice by Directly Affecting the Functional Activity of CD4+CD25+ Regulatory T Cells J. Immunol., February 15, 2007; 178(4): 2458 - 2468. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Yasuda, T. Kuwabara, H. Nakano, K. Aritomi, T. Onodera, M. Lipp, Y. Takahama, and T. Kakiuchi Chemokines CCL19 and CCL21 promote activation-induced cell death of antigen-responding T cells Blood, January 15, 2007; 109(2): 449 - 456. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. D. Keller, P. Rivera Gil, M. Tolle, M. van der Giet, J. Chun, H. H. Radeke, M. Schafer-Korting, and B. Kleuser Immunomodulator FTY720 Induces Myofibroblast Differentiation via the Lysophospholipid Receptor S1P3 and Smad3 Signaling Am. J. Pathol., January 1, 2007; 170(1): 281 - 292. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kabashima, N. M. Haynes, Y. Xu, S. L. Nutt, M. L. Allende, R. L. Proia, and J. G. Cyster Plasma cell S1P1 expression determines secondary lymphoid organ retention versus bone marrow tropism J. Exp. Med., November 27, 2006; 203(12): 2683 - 2690. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. M. L. Meno-Tetang, H. Li, S. Mis, N. Pyszczynski, P. Heining, P. Lowe, and W. J. Jusko Physiologically Based Pharmacokinetic Modeling of FTY720 (2-Amino-2[2-(-4-octylphenyl)ethyl]propane-1,3-diol hydrochloride) in Rats After Oral and Intravenous Doses Drug Metab. Dispos., September 1, 2006; 34(9): 1480 - 1487. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Schmouder, D. Serra, Y. Wang, J. M. Kovarik, J. DiMarco, T. L. Hunt, and M.-C. Bastien FTY720: Placebo-Controlled Study of the Effect on Cardiac Rate and Rhythm in Healthy Subjects. J. Clin. Pharmacol., August 1, 2006; 46(8): 895 - 904. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Fujii, T. Kanai, Y. Nemoto, S. Makita, S. Oshima, R. Okamoto, K. Tsuchiya, T. Totsuka, and M. Watanabe FTY720 suppresses CD4+CD44highCD62L- effector memory T cell-mediated colitis Am J Physiol Gastrointest Liver Physiol, August 1, 2006; 291(2): G267 - G274. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Zemann, B. Kinzel, M. Muller, R. Reuschel, D. Mechtcheriakova, N. Urtz, F. Bornancin, T. Baumruker, and A. Billich Sphingosine kinase type 2 is essential for lymphopenia induced by the immunomodulatory drug FTY720 Blood, February 15, 2006; 107(4): 1454 - 1458. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Sawicka, G. Dubois, G. Jarai, M. Edwards, M. Thomas, A. Nicholls, R. Albert, C. Newson, V. Brinkmann, and C. Walker The Sphingosine 1-Phosphate Receptor Agonist FTY720 Differentially Affects the Sequestration of CD4+/CD25+ T-Regulatory Cells and Enhances Their Functional Activity J. Immunol., December 15, 2005; 175(12): 7973 - 7980. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. I. Singer, M. Tian, L. A. Wickham, J. Lin, S. S. Matheravidathu, M. J. Forrest, S. Mandala, and E. J. Quackenbush Sphingosine-1-Phosphate Agonists Increase Macrophage Homing, Lymphocyte Contacts, and Endothelial Junctional Complex Formation in Murine Lymph Nodes J. Immunol., December 1, 2005; 175(11): 7151 - 7161. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Kharel, S. Lee, A. H. Snyder, S. L. Sheasley-O'Neill, M. A. Morris, Y. Setiady, R. Zhu, M. A. Zigler, T. L. Burcin, K. Ley, et al. Sphingosine Kinase 2 Is Required for Modulation of Lymphocyte Traffic by FTY720 J. Biol. Chem., November 4, 2005; 280(44): 36865 - 36872. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Bandhuvula, Y. Y. Tam, B. Oskouian, and J. D. Saba The Immune Modulator FTY720 Inhibits Sphingosine-1-phosphate Lyase Activity J. Biol. Chem., October 7, 2005; 280(40): 33697 - 33700. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. R Blazar and W. J Murphy Bone marrow transplantation and approaches to avoid graft-versus-host disease (GVHD) Phil Trans R Soc B, September 29, 2005; 360(1461): 1747 - 1767. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Halin, M. L. Scimone, R. Bonasio, J.-M. Gauguet, T. R. Mempel, E. Quackenbush, R. L. Proia, S. Mandala, and U. H. von Andrian The S1P-analog FTY720 differentially modulates T-cell homing via HEV: T-cell-expressed S1P1 amplifies integrin activation in peripheral lymph nodes but not in Peyer patches Blood, August 15, 2005; 106(4): 1314 - 1322. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. A. Vora, E. Nichols, G. Porter, Y. Cui, C. A. Keohane, R. Hajdu, J. Hale, W. Neway, D. Zaller, and S. Mandala Sphingosine 1-phosphate receptor agonist FTY720-phosphate causes marginal zone B cell displacement J. Leukoc. Biol., August 1, 2005; 78(2): 471 - 480. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Davis, J. J. Clemens, T. L. Macdonald, and K. R. Lynch Sphingosine 1-Phosphate Analogs as Receptor Antagonists J. Biol. Chem., March 18, 2005; 280(11): 9833 - 9841. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. G. Lo, Y. Xu, R. L. Proia, and J. G. Cyster Cyclical modulation of sphingosine-1-phosphate receptor 1 surface expression during lymphocyte recirculation and relationship to lymphoid organ transit J. Exp. Med., January 18, 2005; 201(2): 291 - 301. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Han, X. Zhang, G. Wang, H. Guan, G. Garcia, P. Li, L. Feng, and B. Zheng FTY720 suppresses humoral immunity by inhibiting germinal center reaction Blood, December 15, 2004; 104(13): 4129 - 4133. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Allende, T. Sasaki, H. Kawai, A. Olivera, Y. Mi, G. van Echten-Deckert, R. Hajdu, M. Rosenbach, C. A. Keohane, S. Mandala, et al. Mice Deficient in Sphingosine Kinase 1 Are Rendered Lymphopenic by FTY720 J. Biol. Chem., December 10, 2004; 279(50): 52487 - 52492. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 U. Kunzendorf, E. Ziegler, and D. Kabelitz FTY720--the first compound of a new promising class of immunosuppressive drugs Nephrol. Dial. Transplant., July 1, 2004; 19(7): 1677 - 1681. [Full Text] [PDF]
|
|
|
|
 |
|
 K Mayer, F Birnbaum, T Reinhard, A Reis, S Braunstein, F Claas, and R Sundmacher FTY720 prolongs clear corneal allograft survival with a differential effect on different lymphocyte populations Br J Ophthalmol, July 1, 2004; 88(7): 915 - 919. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kimura, A. M. Boehmler, G. Seitz, S. Kuci, T. Wiesner, V. Brinkmann, L. Kanz, and R. Mohle The sphingosine 1-phosphate receptor agonist FTY720 supports CXCR4-dependent migration and bone marrow homing of human CD34+ progenitor cells Blood, June 15, 2004; 103(12): 4478 - 4486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Forrest, S.-Y. Sun, R. Hajdu, J. Bergstrom, D. Card, G. Doherty, J. Hale, C. Keohane, C. Meyers, J. Milligan, et al. Immune Cell Regulation and Cardiovascular Effects of Sphingosine 1-Phosphate Receptor Agonists in Rodents Are Mediated via Distinct Receptor Subtypes J. Pharmacol. Exp. Ther., May 1, 2004; 309(2): 758 - 768. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Allende, J. L. Dreier, S. Mandala, and R. L. Proia Expression of the Sphingosine 1-Phosphate Receptor, S1P1, on T-cells Controls Thymic Emigration J. Biol. Chem., April 9, 2004; 279(15): 15396 - 15401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Bohler, J. Waiser, M. Schuetz, H. H. Neumayer, and K. Budde FTY720 exerts differential effects on CD4+ and CD8+ T-lymphocyte subpopulations expressing chemokine and adhesion receptors Nephrol. Dial. Transplant., March 1, 2004; 19(3): 702 - 713. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Sawicka, C. Zuany-Amorim, C. Manlius, A. Trifilieff, V. Brinkmann, D. M. Kemeny, and C. Walker Inhibition of Th1- and Th2-Mediated Airway Inflammation by the Sphingosine 1-Phosphate Receptor Agonist FTY720 J. Immunol., December 1, 2003; 171(11): 6206 - 6214. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Billich, F. Bornancin, P. Devay, D. Mechtcheriakova, N. Urtz, and T. Baumruker Phosphorylation of the Immunomodulatory Drug FTY720 by Sphingosine Kinases J. Biol. Chem., November 28, 2003; 278(48): 47408 - 47415. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Welsch, S. Hagiwara, J. F. Goetschy, and N. R. Movva Ubiquitin Pathway Proteins Influence the Mechanism of Action of the Novel Immunosuppressive Drug FTY720 in Saccharomyces cerevisiae J. Biol. Chem., July 11, 2003; 278(29): 26976 - 26982. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Yopp, G. J. Randolph, and J. S. Bromberg Leukotrienes, Sphingolipids, and Leukocyte Trafficking J. Immunol., July 1, 2003; 171(1): 5 - 10. [Full Text] [PDF]
|
|
|
|
 |
|
 D. S. Gridley, G. A. Nelson, L. L. Peters, P. J. Kostenuik, T. A. Bateman, S. Morony, L. S. Stodieck, D. L. Lacey, S. J. Simske, and M. J. Pecaut Genetic Models in Applied Physiology: Selected Contribution: Effects of spaceflight on immunity in the C57BL/6 mouse. II. Activation, cytokines, erythrocytes, and platelets J Appl Physiol, May 1, 2003; 94(5): 2095 - 2103. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Fujino, N. Funeshima, Y. Kitazawa, H. Kimura, H. Amemiya, S. Suzuki, and X.-K. Li Amelioration of Experimental Autoimmune Encephalomyelitis in Lewis Rats by FTY720 Treatment J. Pharmacol. Exp. Ther., April 1, 2003; 305(1): 70 - 77. [Abstract] [Full Text]
|
|
|
|
|
|
J. H. Xie, N. Nomura, S. L. Koprak, E. J. Quackenbush, M. J. Forrest, and H. Rosen Sphingosine-1-Phosphate Receptor Agonism Impairs the Efficiency of the Local Immune Response by Altering Trafficking of Naive and Antigen-Activated CD4+ T Cells J. Immunol., April 1, 2003; 170(7): 3662 - 3670. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Brinkmann, M. D. Davis, C. E. Heise, R. Albert, S. Cottens, R. Hof, C. Bruns, E. Prieschl, T. Baumruker, P. Hiestand, et al. The Immune Modulator FTY720 Targets Sphingosine 1-Phosphate Receptors J. Biol. Chem., June 7, 2002; 277(24): 21453 - 21457. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Li, G. M. L. Meno-Tetang, K. Chiba, N. Arima, P. Heining, and W. J. Jusko Pharmacokinetics and Cell Trafficking Dynamics of 2-Amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol Hydrochloride (FTY720) in Cynomolgus Monkeys after Single Oral and Intravenous Doses J. Pharmacol. Exp. Ther., May 1, 2002; 301(2): 519 - 526. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Budde, R. L. Schmouder, R. Brunkhorst, B. Nashan, P. W. Lucker, T. Mayer, S. Choudhury, A. Skerjanec, G. Kraus, and H. H. Neumayer First Human Trial of FTY720, a Novel Immunomodulator, in Stable Renal Transplant Patients J. Am. Soc. Nephrol., April 1, 2002; 13(4): 1073 - 1083. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Bai, J. Liu, Y. Wang, S. Honig, L. Qin, P. Boros, and J. S. Bromberg L-Selectin-Dependent Lymphoid Occupancy Is Required to Induce Alloantigen-Specific Tolerance J. Immunol., February 15, 2002; 168(4): 1579 - 1589. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Hammond-McKibben, P. Lake, J. Zhang, N. Tart-Risher, R. Hugo, and M. Weetall A High-Capacity Quantitative Mouse Model of Drug-Mediated Immunosuppression Based on Rejection of an Allogeneic Subcutaneous Tumor J. Pharmacol. Exp. Ther., June 1, 2001; 297(3): 1144 - 1151. [Abstract] [Full Text]
|
|
|
|
 |
|
 T. Miyamoto, A. Matsumori, M.-W. Hwang, R. Nishio, H. Ito, and S. Sasayama Therapeutic effects of FTY720, a new immunosuppressive agent, in a murine model of acute viral myocarditis J. Am. Coll. Cardiol., May 1, 2001; 37(6): 1713 - 1718. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Nagahara, M. Ikekita, and T. Shinomiya Immunosuppressant FTY720 Induces Apoptosis by Direct Induction of Permeability Transition and Release of Cytochrome c from Mitochondria J. Immunol., September 15, 2000; 165(6): 3250 - 3259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Sun, R. Taniguchi, D. Tanoue, T. Yamaji, H. Takematsu, K. Mori, T. Fujita, T. Kawasaki, and Y. Kozutsumi Sli2 (Ypk1), a Homologue of Mammalian Protein Kinase SGK, Is a Downstream Kinase in the Sphingolipid-Mediated Signaling Pathway of Yeast Mol. Cell. Biol., June 15, 2000; 20(12): 4411 - 4419. [Abstract] [Full Text]
|
|
|
|
|
|
D. D. Pinschewer, A. F. Ochsenbein, B. Odermatt, V. Brinkmann, H. Hengartner, and R. M. Zinkernagel FTY720 Immunosuppression Impairs Effector T Cell Peripheral Homing Without Affecting Induction, Expansion, and Memory J. Immunol., June 1, 2000; 164(11): 5761 - 5770. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Yanagawa, K. Sugahara, H. Kataoka, T. Kawaguchi, Y. Masubuchi, and K. Chiba FTY720, a Novel Immunosuppressant, Induces Sequestration of Circulating Mature Lymphocytes by Acceleration of Lymphocyte Homing in Rats. II. FTY720 Prolongs Skin Allograft Survival by Decreasing T Cell Infiltration into Grafts But Not Cytokine Production In Vivo J. Immunol., June 1, 1998; 160(11): 5493 - 5499. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Mandala, R. Hajdu, J. Bergstrom, E. Quackenbush, J. Xie, J. Milligan, R. Thornton, G.-J. Shei, D. Card, C. Keohane, et al. Alteration of Lymphocyte Trafficking by Sphingosine-1-Phosphate Receptor Agonists Science, April 12, 2002; 296(5566): 346 - 349. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
, FTY720 at 1 mg/kg
p.o. Each symbol represents the mean ± SE of four animals.
Statistical significance was calculated by Dunnett’s test (*:
p < 0.05, **: p < 0.01 vs
vehicle-treated control group).
