The Journal of Immunology, 1999, 163: 2284-2290.
Copyright © 1999 by The American Association of Immunologists
B7.2 (CD86) But Not B7.1 (CD80) Costimulation Is Required for the Induction of Low Dose Oral Tolerance1
Liming Liu,
Vijay K. Kuchroo and
Howard L. Weiner2
Center for Neurologic Diseases, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115
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Abstract
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Oral administration of Ag leads to systemic unresponsiveness (oral
tolerance) to the fed Ag. Oral tolerance is mediated through active
suppression by Th2 or TGF-ß-secreting cells or clonal
anergy/deletion, depending on the Ag dose used, with low dose favoring
active suppression and high dose favoring anergy/deletion. The nature
of APC and inductive events leading to the generation of oral tolerance
have not been well defined. To determine the role of costimulatory
molecules in the induction of oral tolerance, we have tested the effect
of anti-B7.1 or anti-B7.2 mAb on the induction of tolerance by
both high and low dose Ag feeding regimens. Our results show that the
B7.2 molecule is critical for the induction of low-dose oral tolerance.
Injection of anti-B7.2 but not anti-B7.1 intact Ab or Fab
fragments inhibited the oral tolerance induced by low-dose (0.5 mg) but
not high-dose OVA (25 mg) feeding. In addition, anti-B7.2, but not
anti-B7.1, inhibited secretion of TGF-ß, one of the primary
cytokines that mediates low-dose oral tolerance. Finally, in the in
vivo model of experimental allergic encephalomyelitis, anti-B7.2
mAb treatment abrogated protection offered against disease by low-dose
myelin basic protein feeding, while anti-B7.1 had no effect. Anti
B7.2 had no effect on disease suppression by high-dose oral Ag. These
data demonstrate that B7.2 costimulatory molecules play an essential
role in the induction of low-dose oral
tolerance.
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Introduction
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Oral
tolerance is a long-recognized method for inducing peripheral immune
tolerance and is mediated through either active suppression by Th2 or
TGF-ß-secreting cells or by T cell clonal anergy/deletion, depending
on the Ag dose used (1, 2, 3). Low-dose feeding induces
active suppression (1, 2), whereas high-dose feeding
induces T cell clonal anergy or deletion (3). Although the
effector mechanisms of oral tolerance have been elucidated, the
mechanisms associated with the induction phase of oral tolerance are
not well defined. Tolerance mediated by low-dose feeding requires
induction and expansion of Ag-specific regulatory T cells
(4). It is well established that induction and activation
of T cells requires two signals: one is the engagement of the TCR via
the MHC/Ag complex, another comes from a costimulatory signal.
Engagement of CD28 by B7.1 and B7.2 can provide a potent costimulatory
signal (5, 6). In this report, we have investigated the
role of B7 molecules on the induction of oral tolerance by using
anti-B7.1 (1G10) and anti-B7.2 (2D10 and GL1) mAbs. We found
that B7.2 molecules are essential for the induction of low-dose oral
tolerance since anti-B7.2, but not anti-B7.1 mAb or their Fab
fragments, inhibited generation of regulatory T cells. Furthermore,
anti-B7.2 Ab treatment abrogated protective effects of low-dose
oral tolerance in the autoimmune disease model, experimental allergic
encephalomyelitis (EAE)3 (2).
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Materials and Methods
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Animals
Six- to ten-week-old female SJL mice and (PLJ x
SJL)F1 mice were purchased from The Jackson Laboratory (Bar
Harbor, ME) and housed in the animal facility of Harvard Institute of
Medicine (Boston, MA).
Antigens
OVA was purchased from Sigma (St. Louis, MO). Guinea myelin
basic protein (MBP) was prepared from spinal cords using the method of
Diebler et al. (7).
Immunization and EAE induction
For experiments using IFN-
and IL-2 production and induction
of TGF-ß as a readout for oral tolerance, female SJL mice (four to
six per group) were immunized at two sites over flank with a total
volume of 200 µl containing 100 µg OVA and 200 µg of
Mycobacterium tuberculosis (H37Ra; Difco, Detroit, MI) in
IFA. Fourteen days later, spleen cells were prepared and cytokine
production measured with paired Abs from PharMingen (San, Diego,
MA). For EAE induction, (PLJ x SJL)F1 mice (8–10 per
group) were immunized over the flanks with 100 µg of guinea pig MBP
and CFA containing 200 µg of M. tuberculosis in total
volume of 100 µl. Pertussis toxin (List Biologic Laboratories,
Campbell, CA) at 300 ng in 0.2 ml of PBS was given i.p. at the time of
MBP challenge and again 48 h later. Animals were observed for
signs of EAE and scored as follows: 1, tail paralysis; 2, hind limb
weakness; 3, hind limb paralysis; 4, hind limb plus forelimb paralysis;
5, death.
Cytokine ELISA
Cells were prepared from pooled spleens and cultured in
serum-free medium, X vivo 20 (BioWhittaker, Walkersville, MD), and
supernatants were taken at different times after the onset of culture
at 24 h for IL-2, at 48 h for IFN-, and at 72 h for
TGF-ß. Quantitative ELISAs were performed using paired mAbs according
to the protocols provided by manufacturers (PharMingen). TGF-ß was
determined as total TGF-ß with acid activation, as described
previously (8). ELISA results represent mean values from
triplicate cultures. Four mice were used per group. Experimental
results presented are representative of two to three individual
experiments.
Antibodies
1G10 (anti-B7.1), 2D10 (anti-B7.2), and GL-1
(anti-B7.2) were purified from acites by TSD BioServices
(Germantown, NY). Similar results were obtained with both anti-B7.2
Abs. Fab fragments were prepared from 1G10 and 2D10 by TSD BioServices.
Control isotype Abs (rat IgG2a and IgG2b) were purchased from
PharMingen.
Administration of anti-B7 Abs and induction of oral tolerance
In the experiments in which cytokine production was measured,
mice were fed with OVA (25 mg or 0.5 mg) for 5 consecutive days, and
anti-B7.1, anti-B7.2, or control Abs (100 µg in 0.2 ml PBS)
were injected i.p. every other day 30 min before Ag feeding. Mice were
immunized 3 days after the last feeding. In the experiments using EAE
induction as a readout system, MBP (0.5 mg) was fed for 6 consecutive
days, and Abs were injected i.p. every other day 30 min before feeding
MBP. In high-dose group, MBP (20 mg) was fed once and Abs were given
i.p. 30 min before feeding. Slightly lower doses were used for
high-dose MBP feeding, as compared with OVA feeding (20 mg vs 25 mg).
Immunization was conducted 7 days after the last feeding. The
experimental design is depicted in Fig. 1
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FIGURE 1. Flow chart demonstrating experimental design. A,
Experimental design for oral tolerance as measured by cytokine
production. B, Experimental design for oral tolerance in
the EAE model.
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Statistical analysis
Statistics analysis was performed using Student’s
t test.
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Results and Discussion
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We have previously shown that the effector phase of oral tolerance
following low-dose Ag feeding is related to a decrease in Th1 cytokines
(IL-2 and IFN-) and to the induction of TGF-ß-secreting T cells.
Thus, low-dose MBP or hen egg lysozyme feeding induced Ag-specific
TGF-ß-secreting T cells (2, 9). The costimulatory
molecules associated with the induction of oral tolerance and
TGF-ß-secreting cells are not known. To investigate this, we
administered anti-B7.1, anti-B7.2, or control Abs during oral
administration of Ag and immunized the mice 2–3 days later (Fig. 1
).
After 14 days, spleen cells were prepared and cultured in the presence
or absence of OVA, and supernatants were harvested and tested for
cytokine production. As shown in Fig. 2,
multiple low-dose OVA feedings significantly suppressed IL-2 and
IFN- production, as compared with control PBS-fed mice. When the
low-dose feeding regimen was accompanied by i.p. injection of
anti-B7.2 mAb, the production of IL-2 and IFN- was comparable to
that of control PBS-fed mice, indicating that tolerance induction was
abrogated by anti-B7.2 treatment. No effect was observed in animals
treated with anti-B7.1 mAb. However, when high-dose Ag was fed (25
mg OVA) five times, tolerance was induced in all groups, and no effect
was seen with either anti-B7.2 or anti-B7.1 treatment.
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FIGURE 2. Anti-B7.2 mAb but not anti-B7.1 treatment abrogates oral tolerance
induction by low-dose Ag. SJL mice were injected i.p. with 100 µg of
anti-B7.1 (1G10), anti-B7.2 (2D10 or GL1), or isotype control
Ab (rat IgG2a or IgG2b) in 0.2 ml PBS, followed by intragastric
administration of high (25 mg) or low (0.5 mg) doses of OVA
(A). In additional experiments, groups of mice were fed
PBS or OVA and injected with anti-B7.1 or anti-B7.2 Abs, to
test the effect of injecting anti-B7 Abs on subsequent
immunization. Mice were injected with mAb every other day and fed with
OVA or PBS for 5 consecutive days, as depicted in Fig. 1B. Three days after the last feeding, mice were
immunized with OVA plus CFA, as described in Materials and
Methods. Fourteen days after immunization, spleen cells were
prepared and cultured in the presence or absence of OVA for 24 h,
and supernatants were tested for IL-2 and IFN- production by ELISA
assay. The value shown is mean of cytokine production of triplicate
cultures. (Statistics are for both IL-2 and IFN-. Control Ig + fed
PBS vs control Ig + fed 0.5 mg OVA, p < 0.03;
control Ig + fed PBS vs anti-B7.2 + fed 0.5 mg OVA, NS; control
Ig + fed PBS vs anti-B7.1 + fed 0.5 mg OVA, p
< 0.025; control Ig + fed PBS vs anti-B7.2 + fed 25 mg OVA,
p < 0.012; control Ig + fed PBS vs anti-B7.1 +
fed 25 mg OVA, p < 0.016; control Ig + fed PBS vs
anti-B7.1 + fed PBS, NS; control Ig + fed PBS vs anti-B7.2 +
fed PBS, NS.)
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Even though we immunized mice at least 2 days after Ab administration,
it is possible that residual anti-B7.2 Ab could somehow enhance the
effect of immunization, which may account for the observed inhibition
of low-dose oral tolerance induction. To test this, we treated groups
of mice with anti-B7.1 or anti-B7.2 Ab and fed with PBS and
then immunized these mice. As shown in Fig. 2B, these groups
behaved similarly to control Ig-treated and PBS-fed groups, indicating
that anti-B7.2 or anti-B7.1 treatment during the feeding period
did not alter subsquent generation of immune responses. These results
are consistent with previous data that showed the in vivo effects of
the Ab are lost within 48 h (10).
We then investigated the effect of anti B7.1 and anti B7.2 on the
induction of TGF-ß-secreting cells following low-dose feeding. As
shown in Fig. 3A,
anti-B7.2 treatment inhibited the induction of TGF-ß secretion
following low-dose feeding. In contrast, anti-B7.1 Ab treatment did
not significantly affect TGF-ß-secreting cells production. No TGF-ß
production was observed following high-dose feeding, and anti-B7.1
and anti-B7.2 Ab administration had no effect (Fig. 3B).
These results demonstrate that B7.2 molecules are essential for the
induction of low-dose tolerance but not high-dose tolerance and that
B7.2 molecules may be critical for the secretion of TGF-ß. It is not
known whether this is due to lack of expansion of TGF-ß-producing T
cells or inhibition of TGF-ß secretion by other mechanisms.
Additional control experiments were conducted in which animals were
treated with anti-B7.1 or anti-B7.2 and fed with PBS to rule
out any effect of anti-B7.1 or anti-B7.2 on TGF-ß production.
As shown in Fig. 3C, there was no effect of anti-B7.1 or
anti-B7.2 pretreatment in the PBS-fed group, as compared with the
group treated with control Ig.
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FIGURE 3. Anti-B7.2 mAb treatment inhibits TGF-ß secretion induced by low-dose
Ag feeding. SJL mice were injected i.p. with 100 µg of anti-B7.1
(1G10), anti-B7.2 (2D10 or G1), or isotype control Ab (rat IgG 2a
or IgG2b) in 0.2 ml PBS followed by intragastric administration of high
(25 mg) or low (0.5 mg) (A and B) doses
of OVA. In additional experiments, animals were fed with low-dose OVA
and, as a control, were fed with PBS and then injected with
anti-B7.1 and anti-B7.2 (C). Mice were injected
with mAb every other day and fed with OVA 5 consecutive days, as
depicted in Fig. 1. Three days after the last feeding, mice were
immunized with OVA plus CFA, as described in Materials and
Methods. Fourteen days after immunization, spleen cells were
prepared and cultured in the presence or absence of OVA for 72 h
and supernatant were tested for total TGF-ß production. (Anti-B7.2 +
fed 0.5 mg OVA vs control Ig + fed 0.5 mg OVA, p =
0.003; anti-B7.1 + fed 0.5 mg OVA vs control Ig + fed 0.5 mg OVA,
NS. C, Control Ig fed PBS vs anti-B7.1+ fed PBS, NS;
control Ig fed PBS vs anti-B7.2 + fed PBS, NS.
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It is possible that poor binding of anti-B7.1 Ab or insufficient
amounts of anti-B7.1 Ab treatment in vivo could have accounted for
the failure of anti-B7.1 to affect oral tolerance induction. To
address this, before we performed in vivo oral tolerance experiments,
we tested each batch of anti-B7.1 and anti-B7.2 Ab in vitro to
establish that the Abs strongly and specifically bound Chinese hamster
ovary cells transfected with either B7.1 or B7.2 genes. In addition, we
tested the anti-B7.1 Abs used in the current experiments in vivo at
a dose of 100 µg to establish their ability to block EAE, as we have
reported previously (10). Following this, we then
performed in vivo oral tolerance experiments in which up to 500 µg of
anti-B7.1 Ab was used to treat the animals, and, as shown in Fig. 4, we found that 500 µg per injection
still did not inhibit low-dose oral tolerance induction, whereas 100
µg and 500 µg of the anti-B7.2 mab was effective in abrogating
low-dose oral tolerance. Thus, the failure of anti-B7.1 to block
low-dose oral tolerance induction was not due to insufficient
availability or insufficient blockade.
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FIGURE 4. The effect of different doses of anti-B7 Abs on low-dose oral
tolerance induction. SJL mice were injected i.p. with 100 µg or 500
µg of anti-B7.1, B7.2, or isotype control Ab in 0.2 ml PBS,
followed by intragastric administration of different doses of OVA. The
mAb were injected every other day, and mice were fed with OVA for 5
consecutive days, as depicted in Fig. 1. Three days after the last
feeding, mice were immunized with OVA plus CFA, as described in
Materials and Methods. Fourteen days after immunization,
spleen cells were prepared and cultured in the presence or absence of
OVA for 24 h, and supernatants were tested for IFN- production
by ELISA assay. The values shown are mean of cytokine production of
triplicate cultures. (Anti-B7.1 500 µg + fed 0.5 mg OVA vs control Ig
+ fed PBS, p < 0.003; anti-B7.2 100 µg + fed
OVA 0.5 mg vs control Ig + fed PBS, NS; anti-B7.2 500 µg + fed
OVA 0.5 mg vs control Ig + fed PBS, NS.)
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To rule out the possibility that the effect of anti-B7.2 we
observed on abrogating low-dose oral tolerance was due to activation
through Ab cross-linking, we purified Fab fragments from 1G10
(anti-B7.1) and 2D10 (anti-B7.2) and tested them in a manner
similar to the whole mAb. As shown in Fig. 5, anti-B7.2 but not anti-B7.1
mAb Fab fragments inhibited the induction of low-dose tolerance, but
not high-dose tolerance. Thus, the effect of anti-B7.2 mAb appears
due to blocking the costimulatory molecules rather than activation.
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FIGURE 5. Anti-B7.2 mAb Fab fragments inhibit low-dose oral tolerance induction.
SJL mice were injected i.p. with 100 µg of Fab fragments purified
from 1G10 (anti-B7.1), 2D10 (anti-B7.2), or control Ab (rat
IgG) in 0.2 ml PBS, followed by intragastric administration of
different doses of OVA. Mice were injected with Fab every other day and
fed with OVA 5 consecutive days, as depicted in Fig. 1. Three days
after the last feeding, mice were immunized with OVA plus CFA, as
described in Materials and Methods. Fourteen days after
immunization, spleen cells were prepared and cultured in the presence
or absence of OVA for 24 h, and supernatants were tested for IL-2
and IFN- production. A and B, IL-2
production; C and D, IFN- production.
(Statistics are for both IL-2 and IFN-. Control Fab + fed PBS vs
anti-B7.2 Fab + fed 0.5 mg OVA, NS; control Fab + fed PBS vs
anti-B7.1 + fed 0.5 mg OVA, p < 0.04).
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We and others have previously shown that high-dose oral tolerance is
mediated by deletion or induction of anergy in the responding Ag
reactive T cells (3, 11). Recent studies have demonstrated
that engagement of CTLA4 by B7 molecules is critical for the induction
of anergy (12). If the high-dose oral tolerance was
mediated by induction of anergy, one would expect to abrogate high-dose
oral tolerance by blocking both B7 molecules so that they are incapable
of cross-linking CTLA4. We therefore tested whether blocking both B7.1
and B7.2 molecules by either injecting a mixture of anti-B7.1 plus
anti-B7.2 Abs or by injecting CTLA4-Ig, which binds to
both of the B7 molecules, would abrogate high-dose oral tolerance. As
shown in Fig. 6, the combination of
anti-B7.1 and anti-B7.2 behaved similarly to anti-B7.2 alone in
low-dose oral tolerance induction. However, in the high-dose group, the
combination of these two Abs failed to inhibit the tolerance induction.
As shown in Fig. 7, CTLA4-Ig treatment
abrogated high-dose tolerance to OVA, as measured by IFN-
production, although it did not affect IL-2 production. This is
consistent with previous studies of CTLA4-Ig, which demonstrated that
CTLA4-Ig partially inhibits high-dose oral tolerance induction
(13). On the other hand, CTLA4-Ig treatment completely
abrogated low-dose oral tolerance by reversing both the inhibition of
IFN- and IL-2. Thus, whereas anti-B7.2 completely abrogates
low-dose oral tolerance, high-dose tolerance is not abrogated by
administration of anti-B7.1, anti-B7.2, or both of these Abs
given together, although CTLA4-Ig abrogated the IFN- component of
high-dose oral tolerance. The reason for this difference is not known,
but most probably relates to the fact that the costimulatory signal
blockade provided by the combination of anti-B7.1 and anti-B7.2
in terms of affinity and on/off rates is different from that provided
by CTLA4-Ig.
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FIGURE 7. Effect of CTLA4-Ig on high-dose oral tolerance. SJL mice (six mice per
group) were injected i.p. with 200 µg of mouse CTLA4-Ig (Genetics
Institute, Cambridge, MA) or control Ig (PharMingen) in 0.2 ml PBS,
followed by intragastric administration of different doses of OVA or
PBS (low dose, 0.5 mg; high dose, 25 mg). Mice were injected every
other day and fed with OVA 5 consecutive days, as depicted in Fig. 1.
Three days after last feeding, mice were immunized with OVA plus CFA,
as described in Materials and Methods. Fourteen days
after immunization, spleen cells were prepared and cultured in the
presence or absence of OVA for 42 h, and supernatant were tested
for IL-2 and IFN- production. (CTLA4- Ig + fed PBS vs
CTLA4-Ig + fed 25 mg OVA, p = 0.001 for IL-2, NS
for IFN- ; CTLA4-Ig + fed PBS vs CTLA4-Ig + fed 0.5 mg OVA, NS
for both IL-2 and IFN-.)
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To test whether the inhibitory effects of anti-B.7 Abs was
different in the draining and mesenteric lymph nodes as compared with
the spleen, we performed experiments with cells of inguinal and
mesenteric lymph nodes after anti-B7.2 treatment and oral feeding
of Ag. As shown in Fig. 8, we found that
the oral tolerance and its abrogation by anti-B7.2 that we observed
in the spleen was also observed in both the inguinal and mesenteric
lymph nodes.
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FIGURE 8. The effect of anti-B7.2 on the immune response in the inguinal and
mesenteric lymph nodes. SJL mice were injected i.p. with 100 µg of
anti-B7.1 (1G10), B7.2 (2D10 or GL1), or isotype control Ab (rat
IgG 2a or IgG2b) in 0.2 ml PBS, followed by intragastric administration
of different doses of OVA. Mice were injected with mAb every other day
and fed with OVA for 5 consecutive days, as depicted in Fig. 1. Three
days after the last feeding, mice were immunized with OVA plus CFA, as
described in Materials and Methods. Ten days after
immunization, lymph node cells were prepared from the draining lymph
nodes and mesenteric lymph nodes and cultured in the presence or
absence of OVA for 24 h, and supernatants were tested for IFN-
production by ELISA assay. The value shown is mean of cytokine
production of triplicate cultures. (Statistics are for both IL-2 and
IFN-. Control Ig + fed PBS vs anti-B7.2 + fed 0.5 mg OVA,
p = NS in spleen or lymph node; control Ig + fed
PBS vs control Ig + fed 0.5 mg OVA, p = 0.02 for
spleen and 0.01 for lymph node).
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To investigate our findings in an in vivo autoimmune disease model, we
fed (PLJ x SJL) F1 mice with multiple low
doses of MBP (0.5 mg) or a single high dose of MBP (20 mg) and treated
with either anti-B7.1 or anti-B7.2 mAb followed by immunization
with MBP to induce EAE. As shown in Fig. 9, in animals fed multiple low doses of
MBP, treatment with anti-B7.2 mAb abrogated protection offered by
oral feeding of MBP when compared with animals fed low-dose MBP and
treated with control Ig. Consistent with the results obtained with
cytokines, anti-B7.1 did not alter the disease scores of the mice
fed with low-dose MBP, and high-dose tolerance was not affected by the
treatment of either anti-B7.1 or anti-B7.2 Ab. To rule out the
possibility that anti-B7.1 or anti-B7.2 had residual effects on
EAE induction, animals that were fed PBS were treated with
anti-B7.1 and anti-B7.2. As shown in Fig. 10, no effect on EAE was observed.
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FIGURE 9. Anti-B7.2 mAb abrogates suppression of EAE by low-dose MBP
administration. Female (PLJ x SJL) F1 mice (8–10 per
group) were injected i.p. with 100 µg 1G10 (anti-B7.1), 2D10
(anti-B7.2), or isotype control rat Abs in 0.2 ml PBS, followed by
intragastric administration of 0.5 mg of guinea pig MBP. Mice were
injected with mAb every other day and fed for 6 consecutive days. Some
mice were injected with Abs and fed once with 20 mg of guinea pig MBP.
Seven days after last feeding, mice were immunized with guinea pig MBP
and CFA and examined for the onset and severity of EAE. This experiment
was repeated once with similar results. (Control Ig + fed PBS vs
control Ig + fed 0.5 mg MBP, p = 0.0004; control Ig
+ fed PBS vs anti-B7.1 + fed 0.5 mg MBP, p =
0.0003; control Ig + fed PBS vs anti-B7.2 + fed 0.5 mg MBP, NS;
control Ig + fed PBS vs anti-B7.2 + fed 20 mg MBP,
p = 0.0003.)
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FIGURE 10. Anti-B7.1 or anti-B7.2 mAb does not affect disease course in
PBS-fed, MBP-immunized mice. Female (PLJ x SJL) F1
mice were injected i.p. with 100 µg 1G10 (anti-B7.1), 2D10
(anti-B7.2), or isotope control rat Abs in 0.2 ml PBS, followed by
intragastric administration of 0.5 mg of guinea pig MBP or PBS. Mice
were injected with mAb every other day and fed for 6 consecutive days.
Seven days after last feeding, mice were immunized with guinea pig MBP
and CFA and examined for the onset and severity of EAE. (Control Ig +
fed PBS vs control Ig + fed MBP, p = 0.006;
anti-B7.2 + fed MBP vs anti-B7.1 + fed MBP,
p = 0.009; anti-B7.2 + fed PBS vs anti-B7.1
+ fed PBS, NS; control Ig + fed PBS vs anti-B7.1 + fed PBS, NS;
control Ig + fed PBS vs B7.2 + fed PBS, NS.)
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The mechanism by which B7.2 molecules are essential for the induction
of low oral tolerance induction is not clear. B7.2, not B7.1, is the
primary costimulatory molecule responsible for initiating an immune
response (14, 15). Application of anti-B7.2 but not
anti-B7.1 mAb in the induction phase almost completely inhibited
the dinitrofluorobenzene-induced skin contact sensitivity
(16). These data suggest that B7.2 plays dominant role in
the normal functioning of the immune system during primary immune
response (at the time of primary Ag exposure). The reasons for the
dominant role of B7.2 in the initiation of immune response may be
related to the fact that B7.1 and B7.2 are regulated independently,
since, in naive mice, the expression of B7.2 on APCs is significantly
higher than B7.1 (17, 18). Furthermore, B7.2 is rapidly
induced (within 6 h) on activated B cells, whereas B7-1 expression
cannot be detected until 2–3 days after B cell activation
(18).
In terms of B7.2 in the gut, Inaba et al. (19) showed that
Peyer’s patch expressed B7.2 molecules beneath the dome of the
epithelium, the entry site of intestinal Ag. In addition, B cell
follicles and dendritic cells in the interfollicular T cell areas in
the gut expressed B7.2 (19). The expression of B7.1 in
Peyer’s patches has not been reported, presumably due to either lack
of constitutive expression or weak expression. It has been shown that
B7.2 may play a more critical role in the induction of IL-4-producing
cells (10, 19), and IL-4 is a differentiating factor of
TGF-ß-producing Th3 cells (20, 21). In the case of
low-dose Ag feeding, anti-B7.2 Ab may bind to B7.2 molecules on APC
in Peyer’s patches, and thus block the costimulatory signals that are
required for the induction TGF-ß-secreting T cells, leading to the
failure of tolerance induction. Cong et al. (22) have
recently shown that the mucosal adjuvanticity of cholera toxin involves
the selective up-regulation of B7.2 expression, which is consistent
with our finding that B7.2 is important in generation of low-dose oral
tolerance.
In terms of high-dose tolerance, Perez et al. (12) have
recently reported that systemic tolerance induced by i.p. injection of
OVA in IFA was inhibited both by CTLA-4-Ig and anti-CTLA-4,
implying that systemic tolerance is costimulatory-dependent. In a
separate series of experiments, we have found similar costimulatory
requirements for high-dose oral tolerance (13). Of note,
others have recently reported that CD40-CD40L interactions are
important for high-dose tolerance; low-dose tolerance was not tested in
this study (23).
In summary, our findings demonstrate that there are different
costimulatory requirements for low- and high-dose oral tolerance and
that B7.2 plays a critical role in the induction of low-dose oral
tolerance. Whether this is due to quantitatively different signals
generated by B7.1 vs B7.2 or due to difference in the kinetics or level
of expression of these two molecules in the gut remains to be
determined.
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Acknowledgments
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We thank Arlene Sharpe for scientific discussions and Jennifer
Molina for editorial support.
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Footnotes
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1 This work was supported by National Institutes of Health Grant AI43458. 
2 Address correspondence and reprint requests to Dr. Howard L. Weiner, Center For Neurologic Diseases, Brigham and Women’s Hospital, 77 Avenue Louis Pasteur HIM 730, Boston MA 02115-5817. E-mail address:
3 Abbreviations used in this paper: EAE, experimental allergic encephalomyelitis; MBP, myelin basic protein.
Received for publication May 18, 1998.
Accepted for publication June 3, 1999.
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References
|
|---|
-
Friedman, A., H. Weiner. 1994. Induction of anergy or active suppression following oral tolerance is determined by antigen dosage. Proc. Natl. Acad. Sci. USA 91:6688.[Abstract/Free Full Text]
-
Chen, Y., V. K. Kuchroo, J.-I. Inobe, D. A. Hafler, H. L. Weiner. 1994. Regulatory T-cell clones induced by oral tolerance: suppression of autoimmune encephalomyelitis. Science 265:1237.[Abstract/Free Full Text]
-
Chen, Y., J. Inobe, R. Marks, P. Gonnella, V. K. Kuchroo, H. L. Weiner. 1995. Peripheral deletion of antigen-reactive T-cells in oral tolerance. Nature 376:177.[Medline]
-
Chen, Y., J.-i. Inobe, H. L. Weiner. 1997. Inductive events in oral tolerance in the TCR transgenic adoptive transfer model. Cell. Immunol. 178:62.[Medline]
-
Freeman, G. J., V. A. Boussiotis, A. Anumanthan, G. M. Bernstein, K.-Y. Ke, P. D. Rennert, G. S. Gray, J. G. Gribben, L. M. Nadler. 1995. B7-1 and B7-2 do not deliver identical costimulatory signals, since B7-2 but not B7-1 preferentially costimulates the initial production of IL-4. Immunity 2:523.[Medline]
-
Lenschow, D. J., T. L. Walunas, J. A. Bluestone. 1996. CD28/B7 system of T cell costimulation. Annu. Rev. Immunol. 14:233.[Medline]
-
Diebler, G. E., R. E. Martenson, M. W. Kies. 1972. Large scale preparation of myelin basic protein from central nervous tissue of several mammalian species. Prep. Biochem. 2:139.[Medline]
-
Neurath, M. F., I. Fuss, B. L. Kelsall, D. H. Presky, W. Waegell, W. Strober. 1996. Experimental granulomatous colitis in mice is abrogated by induction of TGF-ß-mediated oral tolerance. J. Exp. Med. 183:2605.[Abstract/Free Full Text]
-
Miller, A., O. Lider, A. B. Roberts, M. B. Sporn, H. L. Weiner. 1992. Suppressor T-cells generated by oral tolerization to myelin basic protein suppress both in vitro and in vivo immune responses by the release of TGF-ß following antigen specific triggering. Proc. Natl. Acad. Sci. USA 89:421.[Abstract/Free Full Text]
-
Kuchroo, V., M. Prabhu Das, J. A. Brown, A. M. Ranger, S. S. Zamvil, R. A. Sobel, H. L. Weiner, N. Nabavi, L. H. Glimcher. 1995. B7-1 and B7-2 costimulatory molecules differentially activate the Th1/Th2 developmental pathways: application to autoimmune disease therapy. Cell 80:707.[Medline]
-
Whitacre, C. C., I. E. Gienapp, C. G. Orosz, D. Bitar. 1991. Oral tolerance in experimental autoimmune encephalomyelitis. III. Evidence for clonal anergy. J. Immunol. 147:2155.[Abstract]
-
Perez, V. L., L. V. Parijs, A. Biuckians, X. X. Zheng, T. B. Strom, A. K. Abbas. 1997. Induction of peripheral T-cell tolerance in vivo requires CTLA-4 engagement. Immunity 6:411.[Medline]
-
Samoilova, E. B., J. L. Horton, H. Zhang, S. J. Khoury, H. L. Weiner, Y. Chen. 1998. CTLA4 is required for the induction of high dose oral tolerance. Int. Immunol. 10:491.[Abstract/Free Full Text]
-
Borriello, F., M. P. Sethna, S. D. Boyd, A. N. Schweitzer, E. A. Tivol, D. Jacoby, T. B. Strom, E. M. S. Simpson, G. J. Freeman, A. H. Sharpe. 1997. B7-1 and B7-2 have overlapping, critical roles in immunoglobulin class switching and germinal center formation. Immunity 6:303.[Medline]
-
Lenschow, D., K. Herold, L. Ree, B. Patel, A. Koons, B. Singh, C. Thompson, J. Bluestone. 1996. CD28/B7 regulation of Th1 and Th2 subsets in the development of autoimmune diabetes. Immunity 5:285.[Medline]
-
Nuriya, S., H. Yagita, K. Okumura, M. Azuma. 1996. The differential role of CD86 and CD80 co-stimulatory molecules in the induction and the effector phases of contact hypersensitivity. Int. Immunol. 8:917.[Free Full Text]
-
Miller, S. D., C. L. Vanderlugt, D. J. Lenschow, J. G. Pope, N. J. Karandikar, M. C. D. Canto, J. A. Bluestone. 1995. Blockade of CD28/B7-1 interaction prevents epitope spreading and clinical relapses of murine EAE. Immunity 3:739.[Medline]
-
Lenschow, D. J., A. I. Sperling, M. P. Cooke, G. Freeman, L. Rhee, D. C. Decker, G. Gray, L. M. Nadler, C. C. Goodnow, J. A. Bluestone. 1994. Differential upregulation of the B7-1 and B7-2 costimulatory molecules following immunoglobulin receptor engagement by antigen. J. Immunol. 153:1990.[Abstract]
-
Inaba, K., M. Witmer-Pack, M. Inaba, K. S. Hathcock, H. Sakuta, M. Azuma, H. Yagita, K. Okumura, P. S. Linsley, S. Ikehara, et al 1994. The tissue distribution of the B7-2 costimulation in mice: abundant expression on dendritic cells in situ and during maturation in vitro. J. Exp. Med. 180:1849.[Abstract/Free Full Text]
-
Inobe, J., A. J. Slavin, Y. Komagata, Y. Chen, L. Liu, H. L. Weiner. 1998. IL-4 is a differentiation factor for transforming growth factor-ß-secreting Th3 cells and oral administration of IL-4 enhances oral tolerance in experimental allergic encephalomyelitis. Eur. J. Immunol. 28:2780.[Medline]
-
Seder, R. A., T. Marth, M. C. Sieve, W. Strober, J. J. Letterio, A. B. Roberts, B. Kelsall. 1998. Factors involved in the differentiation of TGF-ß-producing cells from naive CD4+ T cells: IL-4 and IFN- have opposing effects, while TGF-ß positively regulates its own production. J. Immunol. 160:5719.[Abstract/Free Full Text]
-
Cong, Y., C. T. Weaver, C. O. Elson. 1997. The mucosal adjuvanticity of cholera toxin involves enhancement of costimulatory activity by selective up-regulation of B7.2 expression. J. Immunol. 159:5301.[Abstract]
-
Kweon, M. N., K. Fujihashi, Y. Wakatsuki, T. Koga, M. Yamamoto, J. R. McGhee, H. Kiyono. 1999. Mucosally induced systemic T cell unresponsiveness to ovalbumin requires CD40 ligand-CD40 interactions. J. Immunol. 162:1904.[Abstract/Free Full Text]
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Abstract
Introduction
