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,,
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Departments of Neurology and
Immunology, Juntendo University School of Medicine, Tokyo, Japan;
Core Research for Evolutional Science and Technology of Japan Science and Technology Corp., Tokyo, Japan;
Department of Joint Disease and Rheumatism, Nippon Medical School, Tokyo, Japan;
¶ Third Department of Medicine (Neurology), Shinshu University School of Medicine, Matsumoto, Japan; and
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Department of Surgery, Faculty of Medicine, University of Tokyo, Tokyo, Japan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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production by draining lymph
node cells were not reduced or rather enhanced upon restimulation with
proteolipid protein (139–151) in vitro, and these cells effectively
transferred EAE to naive SJL mice. Flow cytometric analyses showed that
the RM134L treatment inhibited the accumulation of OX40-expressing
CD4+ T cells and the migration of adoptively transferred
CD4+ T cells in the spinal cord. Immunohistochemical
staining showed that OX40L was most prominently expressed on
endothelial cells in the inflamed spinal cord. These results suggest
that the OX40/OX40L interaction plays a critical role for the migration
of pathogenic T cells into the CNS in the pathogenesis of
EAE. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Despite the above progresses in our knowledge about the physiological function of OX40/OX40L, their role in various pathological conditions remains to be determined. It has been shown that OX40+ T cells preferentially accumulated in inflammatory sites associated with various diseases and disease models, including rheumatoid arthritis (RA) (22), inflammatory skin diseases (ISD) (23), graft vs-host disease (GVHD) (24), and experimental autoimmune encephalomyelitis (EAE) (25, 26). In rat EAE models, it has been shown that pathogenic T cells resided in the OX40+ population and that depletion of the OX40+ T cells with anti-OX40 mAb-conjugated immunotoxin effectively ameliorated EAE (27). A more recent study showed that therapeutic administration of soluble OX40-Ig fusion protein at the onset of mouse EAE significantly reduced ongoing signs of disease, possibly due to interference of activation of pathogenic T cells by OX40L+ APC in the CNS (28).
In the present study, we examined the contribution of OX40/OX40L to the pathogenesis of EAE, particularly focusing on the development of pathogenic Th1 cells and their migration into the CNS, by administering a neutralizing anti-OX40L mAb to an actively induced or adoptively transferred EAE model in SJL mice. Our present results suggest that OX40/OX40L may play the most critical role at migration of pathogenic T cells into the CNS, because OX40L was most prominently expressed on EC in inflamed CNS and the anti-OX40L mAb treatment efficiently inhibited the accumulation of OX40+ T cells and the migration of adoptively transferred CD4+ T cells in the CNS. Clinical relevancies of the finding are discussed.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Female SJL/J mice, 6–10 wk old, were purchased from Charles River (Atsugi, Japan), housed in the animal care facility at the Juntendo University School of Medicine, and used according to the institutional guidelines.
Antibodies
A neutralizing mAb against mouse OX40L (RM134L, rat IgG2b) was prepared as described previously (11). Control rat IgG was purchased from Sigma (St. Louis, MO). Purified anti-CD16/32 (2.4G2; FcBlock) mAb, FITC-conjugated anti-CD3 (145-2C11), CD4 (RM4-4), CD8 (53-6.7) mAbs, biotinylated anti-OX40 (MRC OX86) mAb, biotinylated control IgG, and PE-labeled streptavidin were purchased from PharMingen (San Diego, CA). High pressure liquid chromatography-purified proteolipid protein (PLP) peptide (PLP139–151; HSLGKWLGHPDKF) was purchased from Sawady Technology (Tokyo, Japan).
Induction of EAE and mAb treatment
To induce active EAE, female SJL/J mice were immunized s.c. with 100 µg PLP139–151 emulsified in CFA containing 200 µg Mycobacterium tuberculosis H37Ra (Difco, Detroit, MI) at four sites in the flank on day 0. Initial clinical signs of disease were usually observed between day 10 and 15 after immunization. These mice were treated i.p. with various doses of anti-OX40L mAb or control IgG daily from day 0 to day 17 when remission was first observed in the control group.
For adoptively transferred EAE, SJL/J mice were immunized with PLP139–151/CFA and draining lymph nodes (DLNs) were harvested on day 10. DLN cells (5 x 106/well) were then cultured with PLP139–151 (30 µg/ml) for 4 days. Cells were then washed with PBS, and 3 x 107 cells in 0.5 ml PBS were injected i.p. into naive SJL/J mice on day 0. The recipient mice were treated i.p. with 400 µg anti-OX40L mAb or control IgG daily from day -1 to day 10 when all mice in the control group showed clinical signs of disease.
Individual mice were examined daily for clinical signs of neurological deficit scored on a 0–6 scale as follows: grade 0, no abnormality; grade 1, limp tail; grade 2, hind limb weakness (waddling gait); grade 3, partial hind limb paralysis; grade 4, complete hind limb paralysis; grade 5, moribund; grade 6, death. The data are reported as the mean daily clinical score for five animals in each group.
Histopathology
For histological evaluation, representative mice from each group were anesthetized and sacrificed by total body perfusion through the left ventricle with 4% paraformaldehyde in PBS. Spinal cords were dissected out, fixed in phosphate-buffered 10% formalin solution, and embedded in paraffin. Thin sections were then stained with hematoxylin and eosin or with Luxol fast blue.
Immunohistochemistry
Frozen sections of the spinal cord from normal SJL/J mice or the mice with active EAE were stained with anti-OX40L (RM134L) mAb, followed by biotin-conjugated goat anti-rat IgG Ab (Kirkegaard and Perry Laboratories, Gaithersburg, MD) and Vectastain ABC kit (Vector Laboratories, Burlingame, CA), and then diaminobenzidine as a substrate.
In vitro proliferation assay
DLN cells were isolated from
PLP139–151-immunized mice on day 10. Cells were
cultured in 96-well flat-bottom microculture plates at a density of
5 x 105 cells/well in 200 µl RPMI 1640
supplemented with 10% FCS, 2 mM L-glutamine, 1 mM sodium
pyruvate, 50 µM 2-ME, and antibiotics in the presence or absence of
30 µg/ml PLP139–151. To assess proliferative
responses, cultures were pulsed with [3H]TdR
(0.5 µCi/well; Dupont-NEN, Boston, MA) for the last 16 h of the
96-h culture and harvested on a Micro 96 Harvester (Skatron, Lier,
Norway). Incorporated radioactivity was measured on a microplate beta
counter (Micro 
Plus; Wallac, Turku, Finland). To assess cytokine
production, culture supernatants were collected at 24, 48, and 72
h and stored at -80°C until the ELISA.
Quantification of cytokines
Cell-free culture supernatants were subjected to
cytokine-specific ELISA for IL-2, IL-4, IL-10, IFN-, and TNF-
according to the protocol recommended by the manufacturer. All
anti-mouse cytokine mAbs and cytokine standards were obtained from
PharMingen.
Isolation of mononuclear cells (MNC) from spinal cord
Dissected spinal cords were first washed in cold RPMI 1640 to remove contaminating blood cells and then homogenized by passage through a wire mesh. The cells were washed in RPMI 1640, resuspended in 80% Percoll (Pharmacia, Uppsala, Sweden), and then overlaid with 40% Percoll to form a discontinuous gradient. The gradient was centrifuged at 1200 x g for 30 min at room temperature. The cells at the 40–80% interface were collected and washed with PBS.
Immunofluorescence and flow cytometry
DLN cells and spinal cord MNC were first preincubated with
anti-CD16/32 mAb to block nonspecific binding of mAb to FcR and
then incubated with biotinylated anti-OX40 mAb or control IgG and
FITC-conjugated mAb against CD3, CD4, or CD8 for 30 min at 4°C,
followed by PE-conjugated streptavidin. After washing with PBS, the
stained cells (live gated on the basis or forward and side scatter
profiles and propidium iodide exclusion) were analyzed on a FACScan
(Becton Dickinson, San Jose, CA).
Adoptive transfer of CFSE-labeled cells
DLN cells were isolated from PLP139–151-immunized mice on day 10 and then restimulated with PLP139–151 (30 µg/ml) for 3 days. Cells were resuspended at 5 x 107 cells/ml in PBS and incubated for 30 min at 37°C with 10 µM CFSE (Molecular Probes, Eugene, OR). Labeling was terminated by adding 5% final concentration of FCS, and the cells were immediately centrifuged and washed with ice-cold PBS. The CFSE-labeled cells (3 x 107) were injected i.p. into naive SJL/J mice on day 0. The recipient mice were treated i.p. with 400 µg anti-OX40L mAb or control IgG daily from day 0 to day 7 after the transfer. Spinal cord MNC, DLN cells, or spleen cells were isolated from the mice on day 10 and subjected to flow cytometry after staining with PE-conjugated anti-CD4 mAb. The absolute numbers of CFSE+CD4+ T cells in the spinal cord, DLN, and spleen were calculated from the total number of collected MNC and the proportion of CFSE+CD4+ T cells among them.
Statistical analysis
The significance of differences between means was determined using the paired t test and the Mann-Whitney U test. P values of <0.05 were considered as significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To explore the contribution of OX40/OX40L interaction to the
development of EAE, we first administered various doses of a
neutralizing anti-OX40L mAb (RM134L) or control rat IgG to
PLP139–151/CFA-immunized SJL mice, which develop
relapsing-remitting EAE resembling multiple sclerosis (MS) in humans,
from the day of immunization (day 0) to day 17 when the remission was
first observed in the control group. As shown in Fig. 1
A, clinical signs of EAE were
first observed on days 11–13, peaked around day 15 (5/5 incidence),
remitted once around day 19, and then relapsed around day 23 in the
control IgG-treated mice, with disease course and severity comparable
with those in the untreated mice (data not shown). In contrast, the
RM134L treatment ameliorated the clinical signs of EAE in a
dose-dependent manner (Fig. 1A). Low doses (100 or 150
µg/day) were effective in reducing the disease severity at the acute
phase (four of five or three of five incidence at day 15, respectively)
but not at the subsequent relapse, and high doses (250 or 400 µg/day)
were effective at both phases (three of five or three of five incidence
at day 15, respectively). Histological examination showed that spinal
cord from the control IgG-treated mice at the acute phase (day 16) had
massive perivascularly infiltrating MNC, which often extended into the
parenchymal white matter (Fig. 2, A and B). In contrast, such mononuclear
infiltrates were greatly reduced in spinal cord from the RM134L-treated
mice (Fig. 2, C and D). These results indicated a
substantial contribution of OX40L to the pathogenesis of actively
induced EAE.
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B. The control IgG-treated mice exhibited a severe and
chronic disease, which was comparable with that in untreated mice (data
not shown). The RM134L treatment greatly reduced the disease severity
throughout the observation period of 25 days even when the treatment
was ceased at day 10 (Fig. 1B). Histological examination
showed massive perivascular infiltrates in spinal cords from the
control IgG-treated mice at day 12 (Fig. 2, E and
F) but only a few infiltrates in spinal cord from the
RM134L-treated mice (Fig. 2, G and H). These
results indicated a critical contribution of OX40L to later phases of
EAE development, including migration of pathogenic T cells into the CNS
and reactivation of pathogenic T cells in the CNS. Effect of anti-OX40L mAb treatment on the development of pathogenic T cells
To determine the contribution of OX40L to the development of
pathogenic T cells in the actively induced EAE,
PLP139–151-immunized mice were treated with
RM134L or control IgG from the day of immunization (day 0) to day 7.
DLN cells were prepared on day 10, restimulated with
PLP139–151 in vitro, and proliferative response
was assessed by [3H]TdR uptake and cytokine
(IL-2, IL-4, IL-10, IFN-, and TNF-) production was assessed by
ELISA. As represented in Fig. 3A, DLN cells from the
RM134L-treated mice showed PLP-specific proliferative response
comparable with that shown by DLN cells from the control IgG-treated
mice. Although IL-2, IL-4, IL-10, and TNF- were not detectable in
the culture supernatants at 24, 48, and 72 h (data not shown),
production of IFN- was significantly increased in DLN cells from the
RM134L-treated mice as compared with that from the control IgG-treated
mice (Fig. 3B). These results indicated that the RM134L
treatment did not inhibit the priming of PLP-reactive T cells in DLN on
s.c. immunization with PLP139–151/CFA but rather
enhanced the development of IFN- producing Th1 cells, which might be
pathogenic.
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A). This difference was more
clearly observed when the transferred cell number was decreased. As
shown in Fig. 4B, the mice that received 1 x
107 DLN cells from the RM134L-treated mice
developed significantly more severe and chronic disease (four of five
incidence at day 15) as compared with the mice that received 1 x
107 DLN cells from the control IgG-treated
mice (two of fiveincidence at day 15). These results indicated
that OX40L isnot required for the development of pathogenic T cells
in the PLP139–151-induced active EAE and may
play rather a suppressive role against the development of pathogenic
Th1 cells.
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We next examined the contribution of OX40L to the distribution of
OX40+ T cells in DLN and spinal cord.
PLP139–151-immunized SJL mice were treated with
RM134L or control IgG from day 0 to day 14. DLN cells and spinal cord
MNC were prepared on day 14 and the presence of
OX40+ T cells was analyzed by two-color flow
cytometry. As shown in Fig. 5B, a substantial number of
OX40-expressing CD4+ T cells were present in
spinal cord from the mice with active EAE. In contrast, the RM134L
treatment substantially inhibited the distribution of
OX40+ T cells, but not that of
OX40- T cells, in the spinal cord. On the other
hand, OX40-expressing CD4+ T cells were rather
accumulated in DLN of the RM134L-treated mice as compared with the
IgG-treated mice (Fig. 5A). These results indicated that
OX40L plays a critical role in distributing OX40+
pathogenic T cells from DLN to the CNS.
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To more directly determine the role of OX40L in the distribution
of pathogenic T cells, the in vitro restimulated DLN cells were labeled
with fluorescent CFSE and adoptively transferred to naive SJL mice,
which were then treated with RM134L or control IgG from day 0 to day 7.
DLN cells, spleen cells, and spinal cord MNC were prepared on day 10
and the presence of transferred CFSE-labeled CD4+
T cells was analyzed by two-color flow cytometry. As shown in Fig. 6, A and C, the
RM134L treatment greatly reduced the number of
CFSE+ CD4+ T cells that
migrated into the spinal cord as compared with the control IgG
treatment. In contrast, the number of
CFSE+CD4+ T cells in the
DLN and spleen was significantly increased by the RM134L treatment
(Fig. 6, B and D). These results indicated that
the OX40/OX40L interaction is involved in the migration of adoptively
transferred pathogenic T cells into the CNS.
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In the above flow cytometric experiments, we could not find a
significant number of OX40L+ cells in DLN cells
or spinal cord MNC (data not shown). We then performed
immunohistochemical staining of spinal cords from normal or actively
induced EAE mice using RM134L mAb. As shown in Fig. 7B, the most prominent
staining was found on vascular EC, which were closely associated with
perivascular mononuclear infiltrates, in the spinal cord from mice with
active EAE on day 16. Some MNC sparsely distributed in the parenchyma
were also positively stained. In contrast, no significant staining was
observed in the spinal cord from normal mice (Fig. 7A).
These results indicated that OX40L is most prominently expressed on
vascular EC in inflamed spinal cord, which appears to be responsible
for the accumulation of OX40+ pathogenic T cells
in the CNS during the development of EAE.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the present study, we examined the contribution of OX40/OX40L to the
pathogenesis of EAE, especially focusing on the development of
pathogenic Th1 cells and their migration into the CNS, by using a
PLP139–151-induced relapsing-remitting EAE model
in SJL mice. The disease course of this model has been well
characterized by previous studies (29, 30, 31). The actively
induced EAE is initiated by immunization with a
PLP139–151 peptide, which is the dominant
encephalitogenic epitope in the SJL mice resulting in the priming of
PLP139–151-reactive T cells in the DLN. The
primed PLP139–151-reactive T cells then migrated
into the CNS and were reactivated by endogenous
PLP139–151 epitope presented by CNS-resident
APC, resulting in the acute disease which peaked around day 15 (Fig. 1A). This acute phase was followed by a transient remission
around day 19, which seems to result from activation-induced cell death
of PLP139–151-reactive T cells within the CNS
(32). The inflammatory reactions at the acute phase
induces the release of endogenous myelin epitopes from damaged myelin
and activates APC within the CNS, leading to de novo activation of
autoreactive T cells specific for endogenous myelin epitopes distinct
from the initiating PLP139–151 epitope. This
phenomenon, called epitope spreading, has been directly implicated in
the occurrence of relapse around day 24 and progression to chronic
disease (31). We administered anti-OX40L mAb from the
day of immunization (day 0) to day 17 to assess the effect on the
development of acute disease. The anti-OX40L mAb treatment
ameliorated the acute disease in a dose-dependent manner, and higher
doses were also effective in ameliorating the relapse (Fig. 1A). This ameliorating effect on relapse after the cessation
of treatment might be simply due to carry over of a sufficient level of
anti-OX40L mAb at the relapse phase, which inhibited the migration
of pathogenic T cells specific for the relapse-associated myelin
epitopes into the CNS. Alternatively, the effect on relapse might be
secondary to the amelioration of acute myelin damage, which limited the
release of endogenous myelin epitopes and the development of pathogenic
T cells specific for relapse-associated epitopes. Our preliminary
observation that a short term treatment with anti-OX40L mAb at the
remission phase also ameliorated the relapse supports the former
possibility. Weinberg et al. (28) recently reported that
administration of an OX40-Ig fusion protein at the onset of acute
disease (days 12–14) moderately reduced ongoing disease but did not
inhibit the relapse. The ameliorating effect of OX40-Ig on acute
disease is consistent with that of anti-OX40L mAb that we observed.
The failure to prevent relapse might be due to a faster clearance of
human OX40-human IgG fusion protein compared with rat IgG
anti-mouse OX40L mAb. Further studies are needed to determine the
therapeutic utility of anti-OX40L mAb for preventing the relapse
and disease progression.
Recent studies using OX40- or OX40L-deficient mice revealed a
substantial contribution of OX40/OX40L interaction to T cell priming in
vivo under certain conditions (18, 19, 20, 21). Chen et al.
(20) reported that OX40L-deficient mice exhibited an
impaired contact hypersensitivity due to a defect in T cell priming and
IFN- production, whereas delayed-type hypersensitivity induced by
OVA/CFA immunization was not affected. In contract, Murata et al.
(21) reported an impaired T cell priming in
OX40L-deficient mice immunized with OVA/CFA, which resulted in reduced
proliferation and cytokine (IFN-, IL-4, and IL-5) production by DLN
cells on restimulation in vitro. In our present study, the
anti-OX40L mAb treatment did not inhibit the priming of
PLP139–151-reactive T cells in DLN on
immunization with PLP139–151/CFA as estimated by
proliferation of DLN cells in response to restimulation with
PLP139–151 in vitro (Fig. 3A). This
intact priming of PLP139–151-reactive T cells in
DLN might be mediated by costimulatory pathways other than OX40/OX40L,
such as CD28/B7. Consistent with this notion, the proliferative
response of DLN cells to PLP139–151
re-stimulation in vitro was almost completely inhibited by addition of
anti-B7–1/B7–2 mAbs but not by that of anti-OX40L mAb (data
not shown). Furthermore, it has been shown that the CD28-mediated
costimulation played a predominant role in induction of EAE in SJL mice
(33, 34). It is worth noting that IFN- production by
DLN cells in response to PLP139–151
re-stimulation in vitro was rather enhanced by the anti-OX40L mAb
treatment (Fig. 3B). This might be due to accumulation of
OX40+ primed T cells in DLN (Fig. 5A).
Alternatively, OX40L might play a rather suppressive role against the
development of IFN--producing Th1 cells, since we and others have
shown that the OX40/OX40L interaction played a preferential role for
the development of Th2 cells in vivo and in vitro
(15, 16, 17).
Our present results clearly indicated that the anti-OX40L mAb
treatment inhibited the accumulation of OX40+ T
cells in the spinal cord, which was evident at the acute phase in the
control IgG-treated mice (Fig. 5B). Weinberg et al.
(28) also recently reported that
OX40+ T cells accumulated in the brain during the
acute phase of EAE, although they have not examined the effect of
OX40-Ig on this accumulation. It has been shown, in a rat EAE model,
that pathogenic T cells resided in the OX40+
population and that depletion of the OX40+ T
cells in vivo ameliorated EAE (25, 26, 27). We also confirmed
that the OX40+ T cell population, but not the
OX40- T cell population, in the
PLP139–151-stimulated DLN cells causes EAE on
adoptive transfer also in the mouse system (data not shown).
Consistently, DLN cells from the anti-OX40L mAb-treated mice that
contained more OX40+ T cells than those from the
control IgG-treated mice (Fig. 5A) induced more severe
disease on adoptive transfer (Fig. 4). Therefore, the reduction of
OX40+ T cells in the CNS appears to be primarily
responsible for the ameliorating effect of anti-OX40L mAb on EAE.
This reduction of OX40+ T cells in the CNS can be
explained by several possibilities. First, anti-OX40L mAb might
inhibit the migration of OX40+ T cells into the
CNS. Second, anti-OX40L mAb might inhibit the clonal expansion of
OX40+ T cells within the CNS, since Weinberg et
al. (28) showed that OX40-Ig significantly inhibited the
proliferative response of PLP-specific T cells lines to brain APC from
active EAE mice in vitro. Third, anti-OX40L mAb might inhibit the
survival of OX40+ T cells within the CNS, because
the OX40 signal has been implicated in the survival of activated T
cells (35, 36). However, our results indicating the
enhanced accumulation of OX40+ T cells in DLN of
the anti-OX40L-treated mice (Fig. 5A) argue against a
requisite role of OX40L for T cell expansion and survival in the second
and third possibilities. The inhibitory effect of anti-OX40L mAb on
the migration of adoptively transferred CD4+ T
cells into the spinal cord (Fig. 6) and the prominent expression of
OX40L on vascular EC in the spinal cord with active EAE, which was
closely associated with perivascular mononuclear infiltrates (Fig. 7),
strongly suggests that the first possibility is most likely.
By immunohistochemical staining with the anti-OX40L mAb, we found
the expression of OX40L on vascular EC and some MNC sparsely
distributed in the parenchyma in the spinal cord with active EAE (Fig. 7B). Weinberg et al. (28) recently demonstrated
that a substantial part of CD11b+ MNC, i.e.,
macrophages/microglia, from the brain with active EAE was
OX40L+ as estimated by flow cytometric analysis
with OX40-Ig, which may correspond to the OX40L+
MNC in the spinal cord parenchyma we observed. However, our present
immunohistochemical staining clearly indicated a prominent expression
of OX40L on vascular EC, and its close association with perivascular
mononuclear infiltrates strongly suggests that the OX40L molecules
expressed on these cells are responsible for the infiltration of
OX40+ T cells into the spinal cord. It has been
reported that cultured human EC constitutively expressed OX40L, which
directly mediated the adhesion of activated T cells expressing OX40 to
these cells in vitro (14). We also observed that some
murine EC lines constitutively expressed OX40L (unpublished data).
However, immunohistochemical staining of human skin tissues showed that
the expression of OX40L on vascular EC was confined to the inflammatory
lesions (23). We also showed in the present study that
OX40L was expressed on EC in inflamed, but not normal, spinal cord
(Fig. 7). These results suggest that the expression of OX40L on EC in
vivo is not constitutive but inducible in response to inflammatory
conditions. Although the proinflammatory factors responsible for the
induction of OX40L expression on EC in vivo remain to be determined by
further studies, it is highly likely that OX40L expressed on activated
EC plays a critical role in recruiting
OX40+-activated T cells to the inflammatory
sites, thus promoting T cell-mediated inflammatory diseases such as EAE
and MS. In such situations, OX40L expressed on EC may directly mediate
adhesion of OX40+ T cells as shown in the human
system in vitro. Alternatively, OX40L on EC may trigger OX40 signaling
in activated T cells, which may activate
41 integrin that has been reported to
play a critical role in migration of pathogenic T cells into the CNS
(37). However, the expression of
41 and L2
integrins was not changed on
OX40+CD4+ T cells in the
DLN of control IgG- or anti-OX40L-treated mice (data not shown). It
is also possible that the OX40/OX40L-mediated costimulation may be
required for T cells to express some chemokine receptor directing their
migration into inflamed CNS, since it has been shown that the OX40
costimulation induced CXCR5 expression in CD4+ T
cells, which directed these cells to B cell areas in the spleen
(15). Further studies are needed to address these
possibilities.
We here demonstrated that OX40L appears to play a critical role in directing migration of OX40+ pathogenic T cells into inflammatory sites in a murine EAE model. A similar accumulation of OX40+ T cells has been observed in various inflammatory diseases, including RA, inflammatory bowel disease (IBD), ISD, and GVHD (22, 23, 24, 38). In addition, expression of OX40L on EC has been observed in inflammatory lesions of RA, IBD, and ISD (Refs. 23 and 38 ; our unpublished observation). Therefore, OX40L may also contribute to the recruitment of pathogenic T cells in these T cell-mediated inflammatory diseases, and the anti-OX40L treatment may be generally applicable to ameliorate these diseases. Consistent with this notion, a recent paper showed that administration of OX40-Ig ameliorated ongoing IBD in mouse models by inhibiting T cell infiltration into the mucosa (38). Further studies are now under way to examine the effect of anti-OX40L mAb treatment on the development of other T cell-mediated inflammatory diseases, such as collagen-induced arthritis and acute GVHD, in mouse models.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Ko Okumura, Department of Immunology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
3 Abbreviations used in this paper: L, ligand; EAE, experimental autoimmune encephalomyelitis; MS, multiple sclerosis; PLP, proteolipid protein; DC, dendritic cells; EC, endothelial cells; RA, rheumatoid arthritis; IBD, inflammatory bowel disease; ISD, inflammatory skin disease; GVHD, graft-vs-host disease; DLN, draining lymph node; MNC, mononuclear cells.
Received for publication April 14, 2000. Accepted for publication November 16, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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8.2+ T cells within the spinal cord of Lewis rats with autoimmune encephalomyelitis. J. Neurosci. Res. 43:42.[Medline]
CDR3 motifs associated with BP recognition are enriched in OX-40+ spinal cord T cells of Lewis rats with EAE. J. Neurosci. Res. 44:562.[Medline]
41 integrin. Nature 356:63.[Medline]
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J. R. Maxwell, R. Yadav, R. J. Rossi, C. E. Ruby, A. D. Weinberg, H. L. Aguila, and A. T. Vella IL-18 Bridges Innate and Adaptive Immunity through IFN-{gamma} and the CD134 Pathway J. Immunol., July 1, 2006; 177(1): 234 - 245. [Abstract] [Full Text] [PDF]
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S. Salek-Ardakani, T. So, B. S. Halteman, A. Altman, and M. Croft Protein Kinase C{theta} Controls Th1 Cells in Experimental Autoimmune Encephalomyelitis J. Immunol., December 1, 2005; 175(11): 7635 - 7641. [Abstract] [Full Text] [PDF]
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C. S. Schmidt, J. Zhao, J. Chain, D. Hepburn, B. Gitter, G. Sandusky, S. Chintalacharuvu, A. Glasebrook, and S. Na Resistance to Myelin Oligodendrocyte Glycoprotein-Induced Experimental Autoimmune Encephalomyelitis by Death Receptor 6-Deficient Mice J. Immunol., August 15, 2005; 175(4): 2286 - 2292. [Abstract] [Full Text] [PDF]
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 J. Mestas, S. P. Crampton, T. Hori, and C. C. W. Hughes Endothelial cell co-stimulation through OX40 augments and prolongs T cell cytokine synthesis by stabilization of cytokine mRNA Int. Immunol., June 1, 2005; 17(6): 737 - 747. [Abstract] [Full Text] [PDF]
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M. Florido, M. Borges, H. Yagita, and R. Appelberg Contribution of CD30/CD153 but not of CD27/CD70, CD134/OX40L, or CD137/4-1BBL to the optimal induction of protective immunity to Mycobacterium avium J. Leukoc. Biol., November 1, 2004; 76(5): 1039 - 1046. [Abstract] [Full Text] [PDF]
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A. D. Weinberg, D. E. Evans, C. Thalhofer, T. Shi, and R. A. Prell The generation of T cell memory: a review describing the molecular and cellular events following OX40 (CD134) engagement J. Leukoc. Biol., June 1, 2004; 75(6): 962 - 972. [Abstract] [Full Text] [PDF]
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L. A. Stephens, A. N. Barclay, and D. Mason Phenotypic characterization of regulatory CD4+CD25+ T cells in rats Int. Immunol., February 1, 2004; 16(2): 365 - 375. [Abstract] [Full Text] [PDF]
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N. Martin-Orozco, Z. Chen, L. Poirot, E. Hyatt, A. Chen, O. Kanagawa, A. Sharpe, D. Mathis, and C. Benoist Paradoxical Dampening of Anti-Islet Self-Reactivity but Promotion of Diabetes by OX40 Ligand J. Immunol., December 15, 2003; 171(12): 6954 - 6960. [Abstract] [Full Text] [PDF]
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 S. Salek-Ardakani, J. Song, B. S. Halteman, A. G.-H. Jember, H. Akiba, H. Yagita, and M. Croft OX40 (CD134) Controls Memory T Helper 2 Cells that Drive Lung Inflammation J. Exp. Med., July 21, 2003; 198(2): 315 - 324. [Abstract] [Full Text] [PDF]
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 T. Totsuka, T. Kanai, K. Uraushihara, R. Iiyama, M. Yamazaki, H. Akiba, H. Yagita, K. Okumura, and M. Watanabe Therapeutic effect of anti-OX40L and anti-TNF-alpha MAbs in a murine model of chronic colitis Am J Physiol Gastrointest Liver Physiol, April 1, 2003; 284(4): G595 - G603. [Abstract] [Full Text] [PDF]
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X. Yuan, A. D. Salama, V. Dong, I. Schmitt, N. Najafian, A. Chandraker, H. Akiba, H. Yagita, and M. H. Sayegh The Role of the CD134-CD134 Ligand Costimulatory Pathway in Alloimmune Responses In Vivo J. Immunol., March 15, 2003; 170(6): 2949 - 2955. [Abstract] [Full Text] [PDF]
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L. Taylor, M. Bachler, I. Duncan, S. Keen, R. Fallon, C. Mair, T. T. McDonald, and H. Schwarz In vitro and in vivo activities of OX40 (CD134)-IgG fusion protein isoforms with different levels of immune-effector functions J. Leukoc. Biol., September 1, 2002; 72(3): 522 - 529. [Abstract] [Full Text] [PDF]
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 A. Yamada, A. D. Salama, and M. H. Sayegh The Role of Novel T Cell Costimulatory Pathways in Autoimmunity and Transplantation J. Am. Soc. Nephrol., February 1, 2002; 13(2): 559 - 575. [Full Text] [PDF]
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L. C. Ndhlovu, N. Ishii, K. Murata, T. Sato, and K. Sugamura Critical Involvement of OX40 Ligand Signals in the T Cell Priming Events During Experimental Autoimmune Encephalomyelitis J. Immunol., September 1, 2001; 167(5): 2991 - 2999. [Abstract] [Full Text] [PDF]
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