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Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Previous experimental findings have shown that treatment of young NOD
mice with anti-IL-10 mAb prevents insulitis (3) and
that the expression of IL-10 in pancreatic ß cells precedes insulitis
(4). These findings implicate that IL-10 as an
immunostimulatory factor in IDDM of NOD mice. BALB/c mice expressing an
IL-10 transgene in their insulin-producing ß cells (IL-10-BALB/c
mice) did not develop diabetes (3), but their offspring
(IL-10-NOD mice) from backcrosses (N2-N3) to NOD mice became diabetic
at an accelerated rate (5). Similarly, NOD mice expressing
IL-10 in the glucagon-producing 
cells of the pancreas experienced
an acceleration in the onset of diabetes (4). Our recent
studies have demonstrated that promotion of diabetes by IL-10 in NOD
mice requires autoreactive T cells but not B cells (6).
Depending upon the circumstances, pancreatic IL-10 supported autoimmune
diabetes either via CD4 or CD8 T cell pathway (6, 7).
Additionally, pancreatic IL-10 promoted insulitis and diabetes in NOD
mice independently of several death-inducing or -signaling molecules
such as Fas, perforin, TNFR-1, and TNFR-2 (8). Taken
together, these results demonstrate that pancreatic expression of
IL-10, as opposed to its expression in the periphery by systemic
administration, enhances pancreatic islet inflammation and loss of
peripheral tolerance leading to accelerated diabetes of the NOD mouse.
In that context, we also found that susceptibility to
cyclophosphamide-induced autoimmune diabetes correlates with pancreatic
islet inflammation (8). That is, Fas-deficient,
insulitis-free NOD.lpr/lpr mice completely resisted
cyclophosphamide-induced diabetes, whereas their littermate
NOD.Fas/lpr mice readily succumbed to autoimmune
diabetes.
Yet the mechanism(s) by which IL-10 promotes autoimmune insulitis and diabetes remains unknown. Therefore, in this study, we have examined the mechanism(s) of IL-10-accelerated diabetes in NOD mice. We examined here, in an adoptive transfer model, whether T cell exposure to IL-10 in the islets in the effector phase of diabetes would counterregulate their diabetogenic potential. Using the IL-10-deficient NOD splenocytes, we also examined whether pancreatic IL-10 alone, without IL-10 produced by autoreactive lymphocytes, is sufficient to drive accelerated autoimmune diabetes. Finally, we tested the role of the adhesion/costimulation (ICAM-1/LFA-1) pathway in IL-10-mediated diabetes of the NOD mouse.
Our results demonstrated, first, that exposure to IL-10 during the effector phase in the islet environment does not inhibit the diabetogenic susceptibility of the T cells because adoptive transfer of splenocytes from diabetic NOD mice into IL-10-NOD.scid/scid mice readily promoted diabetes. Second, pancreatic IL-10 accelerates the transfer of diabetes by splenocytes from prediabetic NOD mice, demonstrating that the compartment of exposure, but not the timing, confers proinflammatory effects on these molecules. Third, adoptively transferring IL-10-deficient NOD splenocytes into IL-10-NOD.scid/scid mice revealed that pancreatic IL-10, but not endogenous IL-10 produced by lymphocytes, is sufficient to promote the diabetes of NOD mice. Fourth, pancreatic IL-10 hyperinduced ICAM-1 expression on vascular endothelium. Fifth, when the ICAM-1 deficiency was introduced into IL-10-NOD and NOD mice, accelerated as well as spontaneous insulitis and diabetes were prevented. Overall, the findings presented here demonstrate that ICAM-1 is required for spontaneous insulitis and diabetes of NOD mice and for the accelerated diabetes of IL-10-NOD mice.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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NOD/shi mice were part of the rodent breeding colony at The Scripps Research Institute (La Jolla, CA). IL-10-BALB/c mice, which express the IL-10 transgene in their islets under the control of human insulin promoter (5), were backcrossed to NOD/shi mice 10 to 11 times to generate IL-10-NOD mice (6). BDC2.5/NOD mice were a gift from Drs. Diane Mathis and Christophe Benoist (Institut National de la Santé et de la Recherche Médicale, France) (9).
Generation of ICAM-1-deficient NOD mice
ICAM-1 deficient C57BL/6 mice (10) were purchased from The Jackson Laboratory (Bar Harbor, ME). These mice were backcrossed onto NOD mice for four to five generations. The mice were selected for NOD MHC at the N2 backcross level. At N4 backcross, mice were intercrossed to obtain wild-type, heterozygous, and knockout mice. The N4F1 intercross was used to introduce ICAM-1 deficiency into IL-10-NOD mice. Heterozygous mice were typed by neo PCR (www.Jax.org). Then the knockout mice were typed by flow cytometry using anti-ICAM-1 mAb (PharMingen, San Diego, CA).
Generation of IL-10-deficient NOD mice
IL-10-deficient (-/-) C57BL/6 mice (11) (provided by Dr. Ralph Kuhn and Werner Muller, Institute for Genetics, University of Cologne, Germany) that were backcrossed to NOD mice for 9 generations (by Drs. Jonathan Katz and Bo Wang, Cincinnati, OH) were further backcrossed to NOD mice until 10 to 12 generations at the Scripps Research Institute. Mice of N10 to N12 backcross generation mice were intercrossed and were used in the experiments described in the manuscript. The heterozygous mice were intercrossed to generate homozygous (-/-), heterozygous (+/-), and wild-type (+/+) mice (12). The primers used to type mice for the IL-10 gene disruption are as follows (www.jax.org): backward (oIMR086): 5'-GTG GGT GCA GTT ATT GTC TTC CCG-3' (1723-1700 in exon I), forward (oIMR087): 5'-GCC TTC AGT ATA AAA GGG GGA CC-3' (1523-1546) (in intron), and backward (oIMR088): 5'-CCT GCG TGC AAT CCA TCT TG-3' (neocassette). A 200-bp PCR product indicates the wild-type allele, and a 400-bp product indicates the mutant allele.
Generation of IL-10-NOD.scid/scid mice
IL-10-BALB/c mice, which express the IL-10 transgene in their islets under the control of human insulin promoter, were backcrossed to NOD.scid/scid mice for 10- to 11 generations. The IL-10-NOD.scid/scid mice were selected for NOD MHC at N2 backcross level as well as for scid/scid mutation by flow cytometry. IL-10-NOD.scid/scid mice never develop insulitis or diabetes throughout their lifetime (B.B. and N.S., unpublished data).
MHC typing of mice
The presence of I-Aßd was determined on
tail DNA by PCR using the following primer set: 5'-GAT ACA TCT ACA ACC
GGG AGG AG-3' (1148-1170) and 5'-CTG TTC CAG TAC TCG GCG TCT G-3'
(1229-1250). PCR amplification for I-Aßd
yielded a 103-bp product from BALB/c but not NOD mice. The presence of
I-Ed was tested in tail DNA by PCR using the
following primer set: 5'-ATG AGC TCC CAG AAG TCA TGG G-3' and 5'-GGA
GAG ACA GCA GCT CTC AGC-3'. PCR amplification for
I-Ed yielded a 277-bp product from BALB/c but
not from NOD mice.
Lymphocyte proliferation assays
Splenocytes from indicated mice were cultured at 5 x 105 cells/well in 200 µl of serum-free HL-1 medium (BioWhittaker, Walkersville, MD) supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin, 2 x 10-3 M L-glutamine, and 3 x 10-5 M 2-ME. Cells were cultured in 96-well flat-bottom microculture plates (Becton Dickinson, Franklin Lakes, NJ) in the presence of indicated Ags for 5 days (13). The cultures were pulsed with 1 µCi of [3H]TdR/well during the last 18 h of the assay and were later harvested. [3H]TdR uptake was measured in a beta scintillation counter. The results were expressed as a stimulation index, i.e., (mean cpm with Ag)/(mean cpm without Ag). OVA (Sigma, St. Louis, MO) was used as a control Ag. Stimulation index values >3 over the background values were considered significant. Background values correspond to cpm obtained with the splenocyte cultures without GAD65. Recombinant GAD65 was prepared as described earlier (13).
Adoptive transfers
To demonstrate the role of pancreatic IL-10 in driving the accelerated autoimmune diabetes, we injected i.v. IL-10-deficient (-/-) and -sufficient (+/-) NOD splenocytes at 3 x 107/mouse into IL-10-NOD.scid/scid and NOD.scid/scid mice. Mice were monitored for diabetes by measuring blood glucose (BG) levels at weekly intervals.
To demonstrate the role of ICAM-1 molecule in the effector phase of autoimmunity, adoptive transfer experiments were performed. ICAM-1-deficient (-/-) or sufficient (+/-; +/+) N4-N5 backcross generation mice were irradiated (700 rad) and were injected with Con A-activated BDC2.5/NOD splenocytes at 5 x 106/mouse i.v. The mice were monitored for diabetes by measuring BG levels at weekly intervals. For activation of BDC2.5/NOD splenocytes, Con A was used at 5 µg/ml for 1 x 107 cells/ml of complete RPMI 1640 medium containing 10% FBS.
Assessment of diabetes
Starting at 4–5 wk of age, mice were tested for diabetes by weekly or biweekly measurements of BG levels using a one-step Bayer Glucometer Elite (Bayer, Elkhart, IN). Animals were considered diabetic when BG levels were >300 mg/dl. In most instances, BG levels exceeded 500 mg/dl.
Histological analysis
Lymphocytic infiltration of the islets was evaluated on hematoxylin and eosin (H&E)-stained paraffin sections of pancreas taken at several levels throughout the organ (13). Paraffin-embedded pancreas sections were stained with an immunoperoxidase method using polyclonal Abs to porcine insulin and synthetic glucagon, followed by a biotinylated secondary Ab and an avidin-biotin complex as described earlier (6). The ICAM-1 expression on vascular endothelium of frozen pancreatic tissue from age-matched, sex-matched IL-10-NOD and nontransgenic (non-tg) NOD mice was determined using biotinylated hamster Ab against mouse ICAM-1 (PharMingen). As a control IgG, we used biotinylated anti-TNP (hamster IgG, group 1) (PharMingen).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Pancreatic expression of IL-10 accelerated diabetes
(6), whereas systemic administration of IL-10 prevented
diabetes (14, 15) of NOD mice, suggesting that the site of
expression and/or timing of experience determined the pro- or
anti-inflammatory effects of IL-10. To address whether the site of
expression determined the proinflammatory effects of IL-10, we used an
adoptive transfer system using splenocytes from diabetic and
prediabetic NOD mice. The results appear in Table I
. Adoptive transfer of splenocytes from
20-wk-old diabetic NOD mice into IL-10-NOD.scid/scid mice
readily elicited clinical disease with faster kinetics (by a margin of
3 wk) than that observed in NOD.scid/scid mice. The findings
show that exposure of diabetogenic splenocytes to IL-10 in the
pancreatic islets did not prevent their disease-causing potential.
Next, to confirm this conclusion, we adoptively transferred splenocytes
from prediabetic NOD (8-wk-old) mice into
IL-10-NOD.scid/scid mice and control
NOD.scid/scid mice. We found that these splenocytes caused
diabetes beginning at 4 wk of post transfer in
IL-10-NOD.scid/scid recipients. The same sample of
splenocytes caused diabetes in non-tg NOD.scid/scid mice
only after 7 wk posttransfer. Our results suggest that exposure to
IL-10 in the pancreatic environment by autoreactive T cells later in
life precipitates disease pathogenesis rather than preventing disease.
The findings also demonstrate that location of expression, not the
timing of experience, plays a role in conferring a proinflammatory
effect on IL-10 in promoting autoimmune diabetes.
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NOD mice spontaneously develop diabetes beginning at 14–15 wk of
age, and their splenocytes transfer clinical disease into
NOD.scid/scid mice. Because IL-10-NOD mice develop
accelerated diabetes beginning at 4–5 wk of age, we tested whether
their splenocytes transfer disease into NOD.scid/scid mice.
The results appear in Table II. Transfer
of splenocytes from 20-wk-old diabetic NOD mice readily caused disease
beginning at 3 wk post transfer, and 82% of the recipient
NOD.scid/scid mice succumbed to diabetes by 7 wk of age
(n = 6). During the same period of time, transfer of
splenocytes from 4- to 5-wk-old diabetic IL-10-NOD mice into
NOD.scid/scid mice elicited disease beginning at 8 wk post
transfer, and 71% of them became diabetic by 16 wk post transfer
(n = 7). However, splenocytes from diabetic IL-10-NOD
mice readily caused diabetes upon transfer into
IL-10-NOD.scid/scid mice (n = 6) beginning
at 3 wk post transfer. The findings 1) demonstrate that splenocytes
from diabetic IL-10-NOD mice cause clinical disease with distinct
kinetics from those splenocytes from diabetic NOD mice and 2) confirm
that local expression of IL-10 in the islets is necessary for
splenocytes from diabetic IL-10-NOD mice to cause accelerated disease
in NOD.scid/scid mice.
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To address whether tg IL-10 produced in the pancreatic islets is
sufficient to induce acceleration of diabetes in IL-10-NOD mice, we
performed adoptive transfer experiments using splenocytes from
age-matched cohorts of nondiabetic IL-10-deficient (-/-) and
IL-10-sufficient (+/+) NOD mice. As recipients, we used
IL-10-NOD.scid/scid and NOD.scid/scid mice. The
results appear in Table III. Adoptive
transfer of splenocytes from 12-wk-old nondiabetic IL-10-deficient
(-/-) NOD mice into IL-10-NOD.scid/scid mice readily
provoked diabetes at 2 wk post transfer (3 of 7 mice; 43% incidence).
Furthermore, 6 of 7 mice (86% incidence) became diabetic by 4 wk of
transfer. During the same period of time, adoptive transfer of
splenocytes from nondiabetic IL-10-deficient (-/-) NOD mice did not
cause diabetes (n = 6; 0% incidence) in
NOD.scid/scid recipients. We also found that transfer of
splenocytes from IL-10-sufficient (+/+) NOD mice into
IL-10-NOD.scid/scid mice caused diabetes beginning at 2 wk
post transfer (2 of 6; 33%). By 4 wk post transfer, 5 of 6
IL-10-NOD.scid/scid recipient mice (83% incidence)
developed diabetes. During the same period of time, none of the
NOD.scid/scid mice that have received splenocytes from
IL-10-sufficient (+/+) NOD mice developed diabetes (n =
6; 0% incidence). Thus, pancreatic IL-10, independent of IL-10
production by autoreactive lymphocytes, is sufficient to drive
accelerated diabetes.
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Because the ICAM-1/LFA-1 pathway is important for extravasation of
autoreactive lymphocytes to the inflamed sites (10), the
acceleration of autoimmune diabetes in IL-10-NOD mice could result from
hyperinduction of ICAM-1 on the vascular endothelium. Therefore, ICAM-1
expression was examined in the pancreatic tissue of age- and
sex-matched IL-10-NOD mice and non-tg littermate controls. As shown in
Fig. 1, ICAM-1 expression was
up-regulated on the vascular endothelium in the pancreata of 5-wk-old
diabetic IL-10-NOD mice compared with that in the pancreata of
age-matched non-tg NOD mice. This finding confirms previous data in
IL-10-BALB/c mice, where pancreatic expression of IL-10 up-regulated
the expression of ICAM-1 on vascular endothelium (5).
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, the
IL-10-NOD mice that are wild type (+/+) (n = 6) or
heterozygous (+/-) (n = 11) for ICAM-1 expression
readily developed diabetes. A noteworthy observation was that ICAM-1
deficiency in IL-10-NOD mice dramatically affected the incidence of
diabetes over a 12-wk period (0 of 4; 0% incidence) and a 16-wk period
(0 of 5; 0% incidence). During this period of time, littermate non-tg
ICAM-1-deficient-NOD mice also failed to develop diabetes. Similarly, a
separate group of ICAM-1-deficient NOD mice (n = 7)
also failed to develop diabetes over a 24-wk period (data not shown).
These results demonstrate that ICAM-1 expression is required for
spontaneous diabetes as well as for IL-10-accelerated diabetes.
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To understand whether ICAM-1 deficiency abrogated acceleration of
diabetes in IL-10-NOD mice by blocking insulitis, we examined the
lymphocytic infiltration in the pancreata of IL-10-NOD mice that are
heterozygous (+/-) or deficient (-/-) for ICAM-1 molecule. The
degree of insulitis was calculated and is shown in Table V. Representative micrographs of the
pancreata from ICAM-1-deficient IL-10-NOD and NOD mice are shown in
Fig. 2. These findings demonstrate that
the absence of ICAM-1 resulted in a striking reduction of insulitis in
IL-10-NOD mice. Yet, because the extent of peri-insulitis was not
affected, apparently ICAM-1 expression is not required for lymphocytes
to extravasate into the pancreas. However, this adhesion molecule is
required for entry of lymphocytes into the islet compartment.
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To further understand whether the protection observed in
ICAM-1-deficient IL-10-NOD mice is due to defective islet Ag-specific
priming, we assessed the proliferation of splenocytes against graded
doses of GAD65 in vitro. As shown in Fig. 3, splenocytes from female
ICAM-1-sufficient NOD mice (n = 3) responded to GAD65
very well (mean stimulation index at 10 µg/ml = 10.4). In
contrast, splenocytes from ICAM-1-deficient NOD mice (n
= 3) failed to respond to GAD65 in vitro (mean stimulation index at 10
µg/ml = 2.0). Similarly, splenocytes from female
ICAM-1-deficient IL-10-NOD mice (n = 3) also failed to
respond to GAD65 in vitro (mean stimulation index at 10 µg/ml =
2.0). Moreover, GAD65-specific T cells from ICAM-1-deficient (-/-)
IL-10-NOD mice produced insignificant amounts of IL-2 and IFN-
and
the levels were below the limits of detection (data not shown).
Clearly, the ICAM-1-dependent pathway is required for the generation
and/or expansion of GAD65-specific T cells in IL-10-NOD mice.
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Although ICAM-1 deficiency forestalled the GAD65-specific T cell
priming in IL-10-NOD mice, ICAM-1-deficient IL-10-NOD mice developed
peri-insulitis but not diabetes. We reasoned that in the absence of
ICAM-1, autoreactive T cells in IL-10-deficient NOD mice may have used
another pathway to home to the pancreatic tissue, but failed to
penetrate the islets and accelerate the onset of diabetes. To learn
whether ICAM-1 deficiency would have blocked diabetes at an effector
phase of autoimmunity, we performed adoptive transfers by using
preactivated islet-specific BDC2.5 T cells. For this purpose, we
irradiated the recipient ICAM-1-deficient and -sufficient NOD mice.
These recipient mice were injected i.v. with Con A-activated, 8-wk-old
female BDC2.5 splenocytes (5 x 106/mouse).
Mice were monitored for diabetes at weekly intervals. As shown in Fig. 4, ICAM-1-deficient IL-10-NOD
(n = 4) as well as ICAM-1-deficient NOD
(n = 4) recipients of Con A-activated BDC2.5
splenocytes remained free from diabetes. Immunohistochemical analysis
of frozen sections of these recipient mice showed that CD4/Vß4 T
cells are around the islets of Langerhans (peri-insulitis) (data not
shown). In contrast, ICAM-1-sufficient NOD (n = 4)
recipients of Con A-activated BDC2.5 splenocytes became diabetic.
Immunohistochemical analysis on frozen sections of these recipient mice
showed that the CD4/Vß4 T cells penetrated the islets of Langerhans
(insulitis) (data not shown). These data suggest that ICAM-1/LFA-1
interaction also plays a role in the effector phase of spontaneous
diabetes in NOD mice and of accelerated diabetes in IL-10-NOD
mice.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Our findings have shown that tg expression of IL-10 in the pancreatic tissue is adequate to drive the pathogenic autoimmune response in vivo. These results emphasize the power of the tg approach to modify autoimmune responses via local production of cytokines. Furthermore, splenocytes from diabetic IL-10-NOD mice, in comparison to splenocytes from diabetic NOD mice, transferred clinical disease with slower kinetics into NOD.scid/scid mice, but accelerated the timing of diabetes upon injection into IL-10-NOD.scid/scid mice. The disparity in these outcomes may be related to the lower frequency of pathogenic T cells in splenocytes of diabetic IL-10-NOD mice. When frequency is limited, IL-10 could effectively promote accelerated diabetes. Alternatively, pancreatic IL-10 may prime ß cells to undergo apoptosis exerted by islet-reactive T cells.
We and others have speculated upon the disparity of effects of
IL-10 on IDDM in the NOD mouse. Indeed, early exposure to IL-10 in the
pancreatic islets accelerates disease (4, 6, 16). However,
systemic exposure to IL-10 during the later prediabetic phase inhibits
disease (14, 15). This inhibition was attributed to the
induction of a protective Th2 response (15). However, we
have shown here that localized pancreatic IL-10 could not
counterregulate the disease state induced by splenocytes from
prediabetic and diabetic NOD mice when the autoreactive lymphocytes
encountered IL-10 during the later part of life or in later stages of
activation. This result suggests that pancreatic IL-10 and systemic
IL-10 may have distinct avenues of immunoregulatory functions. In fact,
the critical target cell for the disparate effects of the cytokine may
differ in each case. Systemic IL-10 may block disease through a more
generalized mechanism such as suppression of APC function (17, 18), effector Th1 development, or the induction of T cell
apoptosis (19). However, within the pancreas,
IL-10 may only affect CD8 T cells that infiltrate early in the disease
process (6) and/or prime the ß cells for apoptosis via
infiltrating autoreactive CD8 T cells. In this context, works from
Groux et al. have shown that IL-10 could be a stimulatory factor for
CD8 T cells (20, 21). It could be argued that the
accelerated diabetes observed in IL-10-NOD mice may result from the
direct toxic effect of IL-10 on ß cells in situ, akin to the data
observed in TNF--NOD mice (22), because a toxic effect
on ß cells could enhance the priming of autoreactive T cells in the
pancreas or lymph nodes. However, we ruled out this possibility because
we failed to detect apoptotic ß cells in the pancreatic tissue of
IL-10-NOD.scid/scid mice (data not shown).
To define more precisely the proinflammatory effects of pancreatic
IL-10 on autoimmune diabetes with respect to the timing vs location of
experience, it would be necessary to establish an inducible (e.g.,
tetracycline-regulated) gene transcription system where islet-specific
expression of IL-10 could be turned on or off at will. In that regard,
Green and coworkers have recently shown that duration of the
inflammatory signal (TNF-), in conjunction with the B7-1 molecule,
is a major criterion in the breakdown of peripheral tolerance to islet
Ags in C57BL/6 mice not genetically predisposed to developing
autoimmunity (23). If the duration of the TNF- signal
is critical for promoting autoimmune diabetes, it is still unclear why
pancreatically expressed TNF- in two different NOD lines either
accelerates or prevents autoimmune diabetes (22, 24, 25).
Despite its status as an immunosuppressive cytokine, IL-10 transgenically expressed in the islets of Langerhans has up-regulated the expression of ICAM-1 on vascular endothelium and accelerated the onset of diabetes. ICAM-1 is known to play a role in inflammatory responses by promoting the migration of leukocytes into the site of inflammation (10). In this context, as this study shows, ICAM-1 deficiency has effectively blocked diabetes in IL-10-NOD mice, confirming the potential role for ICAM-1 in spontaneous diabetes observed in anti-ICAM-1 mAb-treated NOD mice (26). The protection we observed was accompanied by the accumulation of lymphocytes outside islets but not within them, consequently limiting the accessibility of the islets to destruction by infiltrating lymphocytes. Clearly, from its expression pattern on endothelial cells, ICAM-1 must be a critical traffic signal for circulating lymphocytes, inducing recruitment into the islets. ICAM-1 is not only involved in the extravasation of lymphocytes to the inflamed sites, but also in the formation of the "immunological synapse" (27) and in the effector stages of target cell killing (28). Therefore, ICAM-1 can play a role in the initiation and/or effector phases of autoimmune diabetes. We also observed that Con A-activated splenocyte cultures from ICAM-1-deficient NOD and IL-10-NOD mice had reduced numbers of cell clusters (data not shown). Our data also suggest that ICAM-1 plays a role in the effector stages of autoimmune diabetes as adoptive transfer of activated BDC2.5 T cells failed to elicit diabetes in ICAM-1-deficient IL-10-NOD and NOD mice. The work presented here indicates that induction or expression of ICAM-1 is critical for accelerated diabetes because ICAM-1 deficiency precluded the transfer of disease by preactivated diabetogenic BDC2.5 T cells (9).
From the foregoing results, we demonstrate that ICAM-1 expression is pivotal for the acceleration of diabetes. Additionally, our data document that the location, but not the timing, of IL-10 expression determines its pro- or anti-inflammatory effects on autoimmune diabetes of NOD mice. These findings may provide avenues for immune intervention to halt spontaneous as well as accelerated diabetes.
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Acknowledgments |
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Footnotes |
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2 This is manuscript number IMM-13338.
3 Current address: Autoimmunity and Inflammation, Protein Design Labs Incorporated, 34801 Campus Drive, Fremont, CA 94555.
4 Address correspondence and reprint requests to Dr. Nora Sarvetnick, Mail code: IMM-23, Department of Immunology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037.
5 Abbreviations used in this paper: IDDM, insulin-dependent diabetes mellitus; BG, blood glucose; NOD, nonobese diabetic; tg, transgenic.
Received for publication June 15, 2000. Accepted for publication September 25, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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cells accelerates autoimmune insulitis and diabetes in nonobese diabetic mice. Int. Immunol. 6:1927. in neonatal NOD mice promotes diabetes by enhancing presentation of islet antigens. Immunity 9:733.[Medline]
expression in the development of diabetes. Immunity 12:459.[Medline]
in islets prevents autoimmune diabetes in nonobese diabetic (NOD) mice by preventing the development of auto-reactive islet-specific T cells. J. Exp. Med. 184:1963. promotes diabetes in nonobese diabetic mice by CD154-independent antigen presentation to CD8+ T cells. J. Exp. Med. 191:225.This article has been cited by other articles:
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  L. A. Mitchell, J. Gao, R. V. Wal, A. Gigliotti, S. W. Burchiel, and J. D. McDonald Pulmonary and Systemic Immune Response to Inhaled Multiwalled Carbon Nanotubes Toxicol. Sci., November 1, 2007; 100(1): 203 - 214. [Abstract] [Full Text] [PDF]
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 |
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 F. M. Barbe-Tuana, D. Klein, H. Ichii, D. M. Berman, L. Coffey, N. S. Kenyon, C. Ricordi, and R. L. Pastori CD40-CD40 Ligand Interaction Activates Proinflammatory Pathways in Pancreatic Islets Diabetes, September 1, 2006; 55(9): 2437 - 2445. [Abstract] [Full Text] [PDF]
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M. Battaglia, A. Stabilini, E. Draghici, B. Migliavacca, S. Gregori, E. Bonifacio, and M.-G. Roncarolo Induction of Tolerance in Type 1 Diabetes via Both CD4+CD25+ T Regulatory Cells and T Regulatory Type 1 Cells Diabetes, June 1, 2006; 55(6): 1571 - 1580. [Abstract] [Full Text] [PDF]
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 |
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 J. Cantor and K. Haskins Effector Function of Diabetogenic CD4 Th1 T Cell Clones: A Central Role for TNF-{alpha} J. Immunol., December 1, 2005; 175(11): 7738 - 7745. [Abstract] [Full Text] [PDF]
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R. K. Gregg, J. J. Bell, H.-H. Lee, R. Jain, S. J. Schoenleber, R. Divekar, and H. Zaghouani IL-10 Diminishes CTLA-4 Expression on Islet-Resident T Cells and Sustains Their Activation Rather Than Tolerance J. Immunol., January 15, 2005; 174(2): 662 - 670. [Abstract] [Full Text] [PDF]
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B. Greve, L. Vijayakrishnan, A. Kubal, R. A. Sobel, L. B. Peterson, L. S. Wicker, and V. K. Kuchroo The Diabetes Susceptibility Locus Idd5.1 on Mouse Chromosome 1 Regulates ICOS Expression and Modulates Murine Experimental Autoimmune Encephalomyelitis J. Immunol., July 1, 2004; 173(1): 157 - 163. [Abstract] [Full Text] [PDF]
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 Z. Zhu, D. Stevenson, J. E. Schechter, A. K. Mircheff, T. Ritter, L. Labree, and M. D. Trousdale Prophylactic Effect of IL-10 Gene Transfer on Induced Autoimmune Dacryoadenitis Invest. Ophthalmol. Vis. Sci., May 1, 2004; 45(5): 1375 - 1381. [Abstract] [Full Text] [PDF]
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S. Gregori, N. Giarratana, S. Smiroldo, and L. Adorini Dynamics of Pathogenic and Suppressor T Cells in Autoimmune Diabetes Development J. Immunol., October 15, 2003; 171(8): 4040 - 4047. [Abstract] [Full Text] [PDF]
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G. Matarese, V. Sanna, R. I. Lechler, N. Sarvetnick, S. Fontana, S. Zappacosta, and A. La Cava Leptin Accelerates Autoimmune Diabetes in Female NOD Mice Diabetes, May 1, 2002; 51(5): 1356 - 1361. [Abstract] [Full Text] [PDF]
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Z. Kaya, K. M. Dohmen, Y. Wang, J. Schlichting, M. Afanasyeva, F. Leuschner, and N. R. Rose Cutting Edge: A Critical Role for IL-10 in Induction of Nasal Tolerance in Experimental Autoimmune Myocarditis J. Immunol., February 15, 2002; 168(4): 1552 - 1556. [Abstract] [Full Text] [PDF]
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H. H. Smits, E. C. de Jong, J. H. N. Schuitemaker, T. B. H. Geijtenbeek, Y. van Kooyk, M. L. Kapsenberg, and E. A. Wierenga Intercellular Adhesion Molecule-1/LFA-1 Ligation Favors Human Th1 Development J. Immunol., February 15, 2002; 168(4): 1710 - 1716. [Abstract] [Full Text] [PDF]
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 K. Goudy, S. Song, C. Wasserfall, Y. C. Zhang, M. Kapturczak, A. Muir, M. Powers, M. Scott-Jorgensen, M. Campbell-Thompson, J. M. Crawford, et al. Adeno-associated virus vector-mediated IL-10 gene delivery prevents type 1 diabetes in NOD mice PNAS, November 20, 2001; 98(24): 13913 - 13918. [Abstract] [Full Text] [PDF]
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A. La Cava, B. Balasa, A. Good, K. van Gunst, N. Jung, and N. Sarvetnick H-2D End Confers Dominant Protection from IL-10-Mediated Acceleration of Autoimmune Diabetes in the Nonobese Diabetic Mouse J. Immunol., July 15, 2001; 167(2): 1066 - 1071. [Abstract] [Full Text] [PDF]
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