| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
*
The Jackson Laboratory, Bar Harbor, ME 04609; and
Department of Microbiology/Immunology, University of North Carolina, Chapel Hill, NC 27599
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Subpopulations of hematopoietically derived APC include B lymphocytes, macrophages, and dendritic cells (DC). To partially address which of these APC subpopulations could exert immunotolerogenic defects underlying the development of IDDM, we produced a stock of NOD mice made deficient in B lymphocytes by congenic transfer of an Igµ gene functionally disrupted by homologous recombination (formally designated as Igh6tmiCgn and, here, as Igµnull) (7). It had been reported that B lymphocytes have a greater capacity to induce various immunotolerogenic functions than other APC populations (8, 9, 10). Thus, it was anticipated that the elimination of B lymphocytes would not alter, or might even accelerate, IDDM development in NOD mice. However, we were surprised to find that B lymphocyte-deficient NOD.Igµnull mice are IDDM resistant (7). Similar results were obtained by two other groups that produced B lymphocyte-deficient NOD mice by either congenic transfer of an Igµnull allele or treatment with a µ chain-specific Ab (11, 12). The finding that IDDM is inhibited rather than accelerated in NOD.Igµnull mice indicated that B lymphocytes played a newly identified diabetogenic role that is distinct, but not mutually exclusive, from the APC-controlled tolerogenic defects underlying the original development of ß cell-autoreactive T cells in NOD mice.
This newly identified diabetogenic role for B lymphocytes in NOD mice could be as APC with an unique ability to process and present certain ß cell Ags to autoreactive T cells that have been generated as a consequence of the tolerogenic defects described above. Alternatively, B lymphocytes may contribute to IDDM in NOD mice through their ability to secrete autoantibodies that bind to pancreatic ß cells and subsequently trigger autoreactive T cells through an Ab-dependent cell-mediated cytotoxicity response. The present study was conducted to determine whether B lymphocytes contribute to the development of T cell-mediated autoimmune IDDM in NOD mice through either of these two mechanisms.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
NOD/Lt mice are maintained at The Jackson Laboratory (Bar Harbor, ME) by brother-sister mating. Currently, IDDM develops in 90% of female and 63% of male NOD/Lt mice by one year of age. Derivation of a "speed congenic" N7 backcross stock of B lymphocyte-deficient NOD.Igµnull mice fixed to homozygosity for linkage markers delineating all previously identified Idd loci of NOD origin has been described previously (7). From >900 NOD.Igµnull mice generated to date, only 4 female breeders of >30 wk of age have spontaneously developed IDDM. The previously described stock of T and B lymphocyte-deficient NOD-scid (official designation NOD-Prkdcscid) mice is maintained at the N11 backcross generation (13, 14). Similarly, the previously described congenic stock of MHC class I and CD8+ T cell-deficient NOD.ß2mnull (official designation NOD.ß2mtm1Unc) mice is also maintained at the N11 backcross generation (15). These latter two strains served as progenitors for the previously described stock of NOD mice homozygous for both the scid and ß2mnull mutations (designated NOD-scid.ß2mnull) (16, 17). Mice housed at The Jackson Laboratory were maintained under specific pathogen-free conditions and allowed free access to food (National Institutes of Health diet 31A, Purina, Richmond, IN) and acidified drinking water. In addition, all stocks of scid mice were treated for 3 days/week with trimethoprim-sulfamethoxazole (Sulfatrim, Barre National, Baltimore, MD) in the drinking water. Some experiments used NOD/Lt and NOD.Igµnull mice housed in the animal facility of the Department of Microbiology and Immunology at the University of North Carolina at Chapel Hill. Mice in this facility were maintained under viral pathogen-free conditions and allowed access to National Institutes of Health diet 31A (Purina) and acidified drinking water.
Assessment of diabetes and insulitis development
The indicated mice were monitored for development of glycosuria
with Ames Diastix (kindly supplied by Miles Diagnostics, Elkhart, IN).
Glycosuric values of 
3 were considered diagnostic of diabetes onset.
Pancreases from mice assessed for insulitis development were fixed in
Bouin’s solution and sectioned at three nonoverlapping levels.
Granulated ß cells were stained with aldehyde fuchsin and leukocytes
with a hematoxylin and eosin counterstain. Islets (at least 25/mouse)
were individually scored as follows: 0, no lesions; 1, peri-insular
leukocytic aggregates, usually periductal infiltrates; 2, <25% islet
destruction; 3, >25% islet destruction; and 4, complete islet
destruction. An insulitis score for each mouse was obtained by dividing
the total score for each pancreas by the number of islets examined.
Data are presented as mean insulitis score (MIS) ± SEM for the
indicated experimental group.
Ag preparations
The cloning and preparation of the 65-kDa isoform of the candidate murine ß cell autoantigen glutamic acid decarboxylase (GAD) has been described previously (18). Briefly, the cDNA was engineered to encode six histidine residues at the COOH terminus of the protein. Recombinant murine GAD65 was generated in SF21 cells using a Baculovirus expression system and purified using a Ni2+-conjugated resin (Invitrogen, San Diego, CA). The GAD65 was further purified by preparative SDS-PAGE. The protein was then electroeluted and extensively dialyzed against PBS. An SF21 cell extract purified in an identical manner to the recombinant GAD65 protein is not antigenic to T cells. Keyhole limpet hemocyanin (KLH) (Sigma, St. Louis, MO) was used as a control Ag.
Assessment of Ag-primed T cell responses
The indicated number of NOD or NOD.Igµnull mice were immunized in a hind foot pad with a 50-µl emulsion of CFA containing 20 µg of GAD65. After 10 days, T cells were purified by the previously described panning technique (14) from pooled draining lymph nodes (LN) of mice in each group. Triplicate aliquots of 2.5 x 105 T cells were seeded into flat-bottom 96-well microtiter plates in a final volume of 200 µl of the previously described culture medium (19) containing the indicated concentration of GAD, plus 2.5 x 105 irradiated (2000 rad) splenic leukocytes from NOD or NOD.Igµnull mice as a source of APC. Following a 72-h incubation at 37°C in a 95% air/5% CO2-humidified atmosphere, the cultures were pulsed with 1 µCi/well of [3H]thymidine for an additional 16 h. The cultures were then harvested, and [3H]thymidine incorporation was determined using an LKB Betaplate 1205 system (LKB Instruments, Gaithersburg, MD). Data are presented as mean cpm ± SEM.
In other experiments, the indicated mice were immunized in a hind foot
pad with a 50-µl emulsion of CFA containing 20 µg of GAD65 or KLH.
At 10 days after priming, single-cell suspensions were prepared from
pooled draining LN of two to three mice in each experimental group.
Triplicate aliquots of 5 x 105 LN cells were seeded
into flat-bottom 96-well microtiter plates in a final volume of 200
µl of medium with or without 10 µg/ml of GAD or KLH plus 5 x
105 irradiated (2000 rad) NOD or
NOD.Igµnull splenocytes as an
additional source of APC. Following a 72-h incubation, the cultures
were pulsed with 1 µCi/well of [3H]thymidine for
an additional 16 h. Data are presented as mean 
cpm ± SEM
(calculated from the mean Ag stimulated minus unstimulated responses).
Assessment of APC requirements for preactivated GAD-autoreactive T cell clones from NOD mice
GAD65-specific T cell clones were established by culturing 5 x 106 splenocytes from 4-wk-old unimmunized NOD female mice in a 24-well plate in 1.5 ml of RPMI 1640, 5 x 10-5 M 2-ME, 1 mM sodium pyruvate, 1x nonessential amino acids, 1 mM glutamine, 1.0% NOD serum, and 10 µg/ml of intact murine GAD65 for 7 days. T cells (1 x 106) harvested on a Lympholyte M gradient (Cedarlane Laboratories, Hornby, Ontario, Canada) were cultured with 5 x 106 irradiated (3000 rad) NOD splenocytes in 1.5 ml of the above medium and 10 µg/ml GAD65 in a 24-well plate. After 3 days, the cultures were supplemented with medium containing 20 U/ml murine IL-2 (PharMingen, San Diego, CA) and maintained for an additional 3 days, at which time CD4+ T cells were purified using magnetic bead separation (Miltenyi Biotec, Auburn, CA). GAD65-specific CD4+ T cell clones were established via limiting dilution.
To assess the capacity of APC from NOD vs
NOD.Igµnull mice to activate
GAD65-specific T cell clones, 2.5 x 104 cells
from the GAD65-specific 6E12 T cell clone were cultured with 2.5
x 105 irradiated (3000 rad) splenocytes from NOD or
NOD.Igµnull mice in 0.1 ml of the
above medium containing 10% FBS with or without 10 µg/ml of GAD65
for 72 h in a 96-well plate. T cell proliferation was assessed by
measuring the amount of [3H]thymidine incorporation
following a 16-h pulse (1 µCi/well). Data are presented as mean
cpm ± SEM of triplicate cultures.
Purification of B lymphocytes
B lymphocytes were purified from NOD splenic leukocyte preparations using a streptavidin-conjugated magnetic bead system (Miltenyi Biotec) to deplete T cells and macrophages/granulocytes that had been prestained with biotinylated mAbs specific for CD3 (145-2C11) or Mac-1 (M1/70), respectively. Subsequent FACS analysis using an FITC-conjugated goat polyclonal antiserum specific for mouse Ig (Southern Biotechnology Associates, Birmingham, AL) indicated that >95% of the resulting cell preparation consisted of B lymphocytes.
Effect of Ig reconstitution on IDDM development in NOD.Igµnull mice
Total Ig was precipitated from pooled serum of overtly diabetic NOD females by 40% saturation with (NH4)2SO4. The precipitated Ig was then dialyzed extensively against PBS and quantified by ELISA as described previously (16). Female NOD.Igµnull mice were injected i.p. with 70 µg of this Ig preparation twice weekly between 8 and 20 wk of age and simultaneously monitored for diabetes development. Controls consisted of NOD.Igµnull injected on the same schedule with the PBS vehicle alone. At 20 wk of age, circulating serum Ig levels in both groups were determined by ELISA. Insulitis development was also assessed at this time.
Effect of NOD.Igµnull T cells on NOD B lymphocyte repopulation
In an initial experiment, a group of 6-wk-old NOD.Igµnull female mice were injected i.v. with 3 x 106 purified B lymphocytes from standard NOD donors. Controls consisted of NOD-scid females injected with the same preparation of B lymphocytes. At the indicated timepoints, the proportion of donor B lymphocytes among PBL in the NOD.Igµnull and NOD-scid recipients were assessed by FACS using the FITC-conjugated polyclonal antiserum specific for mouse Ig described above.
In subsequent experiments, 3 x 106 purified B lymphocytes from NOD or NOD.ß2mnull female donors, respectively, were injected i.v. into 6-wk-old NOD-scid and NOD-scid.ß2mnull female mice. At the indicated timepoints, the proportion of donor B lymphocytes among PBL in the NOD-scid and NOD-scid.ß2mnull recipients was assessed by FACS. Total PBL counts were also determined to calculate the total number of donor B lymphocytes per ml of peripheral blood in the NOD-scid and NOD-scid.ß2mnull recipients. After stable B lymphocyte repopulation was achieved, a subset of the NOD-scid and NOD-scid.ß2mnull recipients were injected i.v. with 1 x 107 T cells purified from the spleens of female NOD.Igµnull donors as described previously (14). At weekly intervals thereafter, total numbers of donor B lymphocytes per ml of peripheral blood were compared in NOD-scid and NOD-scid.ß2mnull recipients that had or had not been subsequently injected with NOD.Igµnull T cells.
Generation of mixed bone marrow/B lymphocyte chimeras
Female NOD.Igµnull mice were lethally irradiated (1200 rad) at 4 wk of age and then reconstituted as described previously (20) with 5 x 106 T cell-depleted syngeneic bone marrow cells that had been mixed with 3 x 106 purified NOD B lymphocytes. Control chimeras consisted of NOD.Igµnull females reconstituted with syngeneic bone marrow only. Bone marrow chimeras were then monitored at 21 wk postreconstitution for diabetes development. Pancreases from mice that remained normoglycemic through 21 wk postreconstitution were assessed for insulitis development. In addition, upon diabetes onset or at 21 wk postreconstitution, splenic leukocytes from the bone marrow chimeras were typed by FACS for the presence of B lymphocytes as described above and for CD4+ and CD8+ T lymphocytes with the mAbs GK1.5 and 53-6.72, respectively. Furthermore, at 8 wk postreconstitution, a subset of NOD.Igµnull mice reconstituted with syngeneic bone marrow in the presence or absence of NOD B lymphocytes were assessed for presence of GAD-primed T cell responses as described above.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We hypothesized that, if the production of autoantibodies
represents the primary mechanism by which B lymphocytes contribute to
the development of IDDM, disease resistance would be abrogated in
NOD.Igµnull mice infused with Ig
isolated from overtly diabetic NOD donors. Two groups of
NOD.Igµnull females were injected
twice weekly from 8 to 20 wk of age with either 70 µg of Ig isolated
from overtly diabetic NOD donors or with the PBS vehicle control. By 20
wk of age, ELISA measurements confirmed the presence of circulating Ig
in sera of the experimental (69.2 ± 4.9 µg/ml), but not the
control group (0.9 ± 0.2 µg/ml), of
NOD.Igµnull mice (Table I
). However, IDDM failed to develop in
any of the Ig-reconstituted (0/8) or control (0/8)
NOD.Igµnull female mice.
Furthermore, the level of insulitis in Ig-reconstituted
NOD.Ignull mice (MIS = 0.85
± 0.40) was not significantly different from the low levels observed
in PBS-treated controls (MIS = 1.29 ± 0.36). Thus, the
levels of Ig reconstitution achieved in this experiment failed to
accelerate autoimmune destruction of pancreatic ß cells in
NOD.Ignull mice. This indicates that
the production of autoantibodies is unlikely to represent the primary
mechanism by which B lymphocytes contribute to the development of T
cell-mediated autoimmune IDDM in standard NOD mice.
|
Given that Ig infusions did not abrogate IDDM resistance or
promote insulitis development in NOD.Igµnull
mice, we hypothesized that the pathogenic role for B lymphocytes in
this disease may be as APC with a unique ability to process and present
certain pancreatic ß cell Ags to autoreactive T cells. As an initial
test of this hypothesis, we determined whether NOD and
NOD.Igµnull mice differed in ability to
generate T cell responses to the candidate pancreatic ß cell
autoantigen GAD. While clearly many different ß cell proteins are
recognized by autoreactive T cells in IDDM, we chose GAD as a model
autoantigen for our studies based on reports that it is among the
earliest targets of diabetogenic T cells in NOD mice (18, 21).
NOD.Igµnull mice were characterized
by an absence of spontaneous T cell responses to GAD (data not shown).
However, it was possible that GAD-reactive T cells were still present
but not efficiently activated in
NOD.Igµnull mice because of a
quantitative loss in APC resulting from the absence of B lymphocytes.
We addressed this issue through use of an in vivo priming protocol that
would amplify any GAD-reactive T cells present in either standard NOD
or NOD.Igµnull mice. As expected, T cells from
GAD65-primed NOD mice proliferated when restimulated with this Ag in
vitro in the presence of standard B lymphocyte-positive NOD APC and, to
a lesser extent, in the presence of B lymphocyte-deficient APC from
NOD.Igµnull mice (Fig. 1). In contrast, T cells from
GAD65-primed NOD.Igµnull mice
failed to respond upon antigenic restimulation in vitro in the presence
of either B lymphocyte intact or deficient APC. These results indicated
that the initial in vivo priming of GAD-reactive T cell responses in
NOD mice requires the presence of Ag-presenting B lymphocytes. However,
other APC populations such as macrophages and DC appear to be able to
process and present GAD65, albeit less efficiently than B lymphocytes,
to NOD T cells previously primed against this Ag.
|
The results described above indicated that once activated in a B
lymphocyte-dependent fashion, the response of GAD-reactive T cells from
standard NOD mice can be maintained by other APC subpopulations.
Assuming that this conclusion was correct, we reasoned that B
lymphocyte-deficient APC should stimulate a response by the
GAD-reactive T cell clone 6E12, which was originally propagated from a
standard NOD mouse. As shown in Figure 2,
B lymphocyte-deficient APC from
NOD.Igµnull mice were able to
process and present antigenic peptides from intact GAD65 to the 6E12 T
cell clone, albeit at lower levels than B lymphocyte-intact APC from
standard NOD mice. Similar results were obtained using a peptide
comprising amino acids 216 to 235 from GAD65, which is the antigenic
target of the 6E12 T cell clone (R. M. T., personal
observation). Thus, while B lymphocytes are necessary APC for
initiating GAD-autoreactive T cell responses in NOD mice, once such
effectors are triggered their activities can be maintained by other APC
subpopulations.
|
The failure of NOD.Igµnull
mice to generate a T cell response to GAD suggested the diabetogenic
role of B lymphocytes may be as APC with a preferential ability to
process and present certain MHC class II-restricted ß cell
autoantigens. However, IDDM resistance in
NOD.Igµnull mice could also be
explained if the absence of B lymphocytes results in a generic
inability to generate a T cell response to all, rather than a specific
subset, MHC class II-restricted Ags. In mouse strains other than NOD,
KLH has been reported to be an MHC class II-restricted Ag that does not
require B lymphocytes for presentation to T cells (23). T cells within
LN from both standard NOD and
NOD.Igµnull mice primed with KLH
responded equivalently when restimulated with this Ag in the presence
of B lymphocyte-deficient APC (Fig. 3).
However, the recall response of T cells within LN from both KLH-primed
NOD and NOD.Igµnull mice was
greater in the presence of B lymphocyte-intact than B
lymphocyte-deficient APC. Thus, B lymphocytes are not required for the
initial priming of KLH-reactive T cells in NOD mice but do contribute
to the amplification of such responses. These data demonstrate that the
absence of B lymphocytes in the
NOD.Igµnull stock has only
eliminated their ability to generate T cell responses to certain MHC
class II-restricted Ags.
|
The fact that NOD.Igµnull mice
cannot generate a T cell response against GAD, but can do so to KLH,
suggests that the diabetogenic role of B lymphocytes is as APC with a
preferential ability to process and present certain MHC class
II-restricted ß cell autoantigens. As an initial test of this
hypothesis, we wished to determine whether
NOD.Igµnull mice repopulated with B
lymphocytes were rendered IDDM susceptible and whether this was
associated with a restored ability to generate a T cell response to
GAD. However, purified NOD B lymphocytes only transiently repopulated
unmanipulated NOD.Igµnull
recipients (Fig. 4). These same B
lymphocytes permanently repopulated NOD-scid recipients,
which lack endogenous T as well as B lymphocytes. Thus, we hypothesized
that a population of MHC class I-restricted cytotoxic T cells present
in NOD.Igµnull, but not
NOD-scid, recipients mediates an immunologic rejection of
transplanted B lymphocytes. We were able to test this hypothesis
because of the availability of both standard NOD and
NOD-scid mice made deficient in MHC class I expression by
the presence of a ß2mnull allele
(15, 16, 17). Standard MHC class I-positive NOD B lymphocytes previously
engrafted into NOD-scid recipients were rapidly eliminated
following the subsequent infusion of T cells from
NOD.Igµnull donors (Fig. 5). In contrast, infusion of these same T
cells did not mediate the elimination of MHC class I-negative B
lymphocytes from NOD.ß2mnull
donors that had been previously engrafted into
NOD-scid.ß2mnull
recipients. Thus, NOD.Igµnull mice
harbor a population of T cells that can mediate a MHC class
I-restricted cytotoxic response against standard NOD B lymphocytes.
This is most likely due to the fact that during the course of their
differentiation, T cells in
NOD.Igµnull mice fail to encounter
B lymphocytes and thus are not rendered tolerant to them. It should
also be noted that even following in vivo priming,
NOD.Igµnull mice failed to generate
T cell responses to Ig isolated from standard NOD donors (data not
shown). Thus, Ig does not appear to represent a B lymphocyte protein to
which NOD.Igµnull T cells fail to
establish tolerance. This was not unexpected, since
NOD.Igµnull T cells reject
transplanted NOD B lymphocytes in an MHC class I-restricted fashion,
while any T cell response engendered against soluble Ig would most
likely be MHC class II restricted.
|
|
We hypothesized that T cells from NOD.Igµnull mice could be rendered tolerant to B lymphocytes if forced to mature in their presence. If correct, such an approach would make it possible to determine whether restoring the presence of B lymphocytes reconstitutes an APC function that abrogates IDDM resistance in NOD.Igµnull mice. To address these issues, NOD.Igµnull mice were lethally irradiated to ablate pre-existing immunologic effectors and then reconstituted with purified NOD B lymphocytes admixed with NOD.Igµnull bone marrow as a source of T cell precursors. Controls consisted of lethally irradiated NOD.Igµnull mice reconstituted with syngeneic bone marrow only, as well as NOD-scid recipients repopulated with the purified B lymphocytes alone.
As expected, spleens from
NOD.Igµnull mice reconstituted with
syngeneic bone marrow alone contained both CD4+ and
CD8+ T cells but no B lymphocytes (Table II). IDDM developed in only 1/15 of
NOD.Igµnull mice reconstituted with
syngeneic marrow alone. Insulitis development was also quite limited
(MIS = 1.32 ± 0.33) in
NOD.Igµnull mice that remained free
of overt IDDM following reconstitution with syngeneic bone marrow
alone, indicating minimal activation of diabetogenic effectors. In
contrast, permanent B lymphocyte as well as CD4+ and
CD8+ T cell repopulation was detected in spleens of
NOD.Igµnull mice reconstituted with
syngeneic marrow admixed with purified NOD B lymphocytes. This
indicated that T cells derived from
NOD.Igµnull marrow are rendered
tolerant to B lymphocytes when forced to mature in their presence. IDDM
developed in 15/23 of NOD.Igµnull
mice characterized by chimeric restoration of both T and B lymphocyte
populations. In addition, significant levels of insulitis were present
(MIS = 2.06 ± 0.41) in the few T and B
lymphocyte-repopulated NOD.Igµnull
mice that remained free of overt IDDM. IDDM failed to develop in any
(0/5) NOD-scid females repopulated with the purified B
lymphocytes alone. GAD-reactive T cell responses were also restored in
NOD.Igµnull mice that had been
rendered diabetes susceptible following reconstitution with syngeneic
bone marrow plus NOD B lymphocytes (Fig. 6). Such GAD-reactive T cell responses
remained absent in diabetes-resistant
NOD.Igµnull mice that had been
reconstituted with syngeneic marrow only. Thus, the presence of B
lymphocytes appears to restore IDDM susceptibility to
NOD.Igµnull mice by reconstituting
a normally absent APC function.
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Unlike reconstitution with B lymphocytes, infusion of Ig isolated from
overtly diabetic NOD donors did not abrogate IDDM resistance or enhance
insulitis development in
NOD.Igµnull mice. This suggested
that autoantibody production does not represent the primary
diabetogenic role of B lymphocytes in NOD mice. It should be noted that
the reconstituted levels of circulating Ig achieved in
NOD.Igµnull mice (
70 µg/ml)
were approximately 10-fold less than that of standard NOD mice
(700–1000 µg/ml). Thus, we cannot exclude the possibility that
higher levels of Ig reconstitution might have a diabetogenic effect in
NOD.Igµnull mice. However, findings
from a recent report suggest that if autoantibodies do play a primary
diabetogenic role, the levels of Ig reconstitution we achieved in
NOD.Igµnull mice should have been
sufficient to trigger an observable pathogenic effect (25). This study
found that the Sjogren’s syndrome-like pathology that characterizes
standard NOD mice does not develop in the same B lymphocyte-deficient
NOD.Igµnull stock used in this
study. Interestingly, a Sjogren’s pathology was restored in
NOD.Igµnull mice that received a
single 100 µg injection of Ig isolated from either standard NOD mice
or human patients with this disease. Given these results, it seems
likely that if autoantibodies also play a primary role in IDDM
development, their pathogenic effect would have been observed in
NOD.Igµnull mice injected twice
weekly from 8 to 20 wk of age with 70 µg of Ig isolated from standard
NOD donors. Thus, while contributions from autoantibody production
cannot be completely excluded, our data support the conclusion that the
primary diabetogenic role for B lymphocytes in NOD mice is as a
subpopulation of APC essential to the initiation of certain ß
cell-autoreactive T cell responses.
B lymphocytes represent an essential subpopulation of APC for generating MHC class II-restricted T cell responses to certain Ags (23). It has been reported that an MHC class II-restricted T cell response against the putative ß cell Ag GAD may have a critical early role in triggering a cascade of other autoreactive T cell responses, which ultimately leads to the development of IDDM in NOD mice (18, 21, 26). Thus, GAD was used as a model to test the hypothesis that the diabetogenic role for B lymphocytes in NOD mice is as a subpopulation of APC with a preferential ability to process and present certain MHC class II-restricted ß cell Ags to autoreactive T cells. Unlike standard NOD, B lymphocyte-deficient NOD.Igµnull mice were found to be incapable of generating primed T cell responses against GAD. However, NOD.Igµnull mice could still generate primed T cell responses against the control Ag KLH. Thus, B lymphocytes represent a critical subpopulation of APC for triggering certain, but not all, T cell responses in NOD mice. The fact that GAD-specific T cell responses are among those eliminated in the NOD.Igµnull stock indicates that the diabetogenic role of B lymphocytes in standard NOD mice is as a subpopulation of APC with a preferential ability to initiate T cell reactivity against certain key ß cell autoantigens. Support for this conclusion was provided by the finding that GAD-reactive T cell responses were restored in NOD.Igµnull mice that had been rendered IDDM susceptible by B lymphocyte repopulation. The fact that they are among the earliest cells to infiltrate the pancreatic islets of NOD mice (27, 28) also supports our conclusion that B lymphocytes represent a critical subpopulation of APC for initiating the development of T cell-mediated autoimmune IDDM.
Interestingly, only the initiation of GAD-reactive T cell responses in NOD mice requires B lymphocytes as APC. Once such GAD-reactive T cells have been initially activated in NOD mice, our data indicate that their responses can be maintained by APC other than B lymphocytes. It is also possible that B lymphocytes are necessary APC only at the initiative phases of other MHC class II-restricted ß cell-autoreactive T cell responses. If preactivated ß cell-autoreactive T cells do not require B lymphocytes as APC, this could explain the finding that T cells from overtly diabetic NOD donors transferred disease to young prediabetic recipients depleted of B lymphocytes by treatment with a µ chain-specific Ab (29).
One factor that complicated our ability to assess the diabetogenic function of B lymphocytes was the rejection of these cells upon transfer into unmanipulated NOD.Igµnull recipients. This rejection of B lymphocytes in NOD.Igµnull recipients was mediated by an MHC class I-restricted cytotoxic T cell response. The presence of such effectors appears to result from the fact that during their differentiation, T cells in NOD.Igµnull mice fail to encounter B lymphocytes and thus are not rendered tolerant to them. This was demonstrated by our finding that T cells differentiating from precursors in NOD.Igµnull bone marrow were rendered tolerant to B lymphocytes when forced to mature in their presence. Full IDDM susceptibility was restored to NOD.Igµnull mice that had been permanently repopulated with both T and B lymphocytes by this chimerization technique.
The fact that T cells from unmanipulated NOD.Igµnull are not tolerant to B lymphocytes does not result from the generalized autoimmune proclivity of the NOD strain (30). We base this conclusion on the finding that T cells developing from NOD.Igµnull marrow could be rendered tolerant to B lymphocytes when forced to mature in their presence. This indicates that the induction of tolerance to B lymphocytes is a normal feature in development of the T cell repertoire. The thymic site of T cell differentiation contains very few B lymphocytes. Thus, it seems most likely that T cells that are potentially autoreactive against syngeneic B lymphocytes are deleted or inactivated shortly after their emigration from the thymus. If this supposition is correct, it would also suggest that an immediate encounter with their cognate Ag results in tolerogenic responses, rather than the functional activation of T cells that have recently emigrated from the thymus. Conversely, it has been reported that the small numbers of B lymphocytes that do reside within the thymus may play an important role in the normal induction of T cell tolerance to endogenous Ags (31, 32). Thus, the normal induction of T cell tolerance to B lymphocytes could occur intrathymically. Interestingly, NOD mice are reported to be characterized by increased numbers of intrathymic B lymphocytes (33). However, regardless of whether NOD B lymphocytes induce tolerance to themselves intrathymically or in the periphery, they are clearly unable to block the development or function of pancreatic ß cell-autoreactive T cells.
In conclusion, this study indicates that B lymphocytes play a diabetogenic role in NOD mice as APC essential to the initiation of effector T cell responses against certain pancreatic ß cell autoantigens such as GAD. It remains to be determined whether the initiation of T cell responses to any candidate ß cell autoantigens other than GAD also requires B lymphocytes as APC. However, regardless of their identity or number, our study indicates that any ß cell Ags which initially require B lymphocytes as APC will be among the most pathogenically relevant targets of MHC class II-restricted diabetogenic T cell responses.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Abbreviations used in this paper: NOD, nonobese diabetic; IDDM, insulin-dependent diabetes mellitus; DC, dendritic cells; MIS, mean insulitis score; GAD, glutamic acid decarboxylase; KLH, keyhole limpet hemocyanin; LN, lymph nodes.
Received for publication April 4, 1998. Accepted for publication June 9, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
This article has been cited by other articles:
|
|
 |
|
  P. L. Kendall, D. J. Moore, C. Hulbert, K. L. Hoek, W. N. Khan, and J. W. Thomas Reduced Diabetes in btk-Deficient Nonobese Diabetic Mice and Restoration of Diabetes with Provision of an Anti-Insulin IgH Chain Transgene J. Immunol., November 15, 2009; 183(10): 6403 - 6412. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Truong, W. W. Hancock, J. C. Plester, S. Merani, D. C. Rayner, G. Thangavelu, K. M. Murphy, C. C. Anderson, and A. M. J. Shapiro BTLA targeting modulates lymphocyte phenotype, function, and numbers and attenuates disease in nonobese diabetic mice J. Leukoc. Biol., July 1, 2009; 86(1): 41 - 51. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. H. Smith and T. F. Tedder Targeting B-cells Mitigates Autoimmune Diabetes in NOD Mice: What Is Plan B? Diabetes, July 1, 2009; 58(7): 1479 - 1481. [Full Text] [PDF]
|
|
|
|
|
|
E. Marino, J. Villanueva, S. Walters, D. Liuwantara, F. Mackay, and S. T. Grey CD4+CD25+ T-Cells Control Autoimmunity in the Absence of B-Cells Diabetes, July 1, 2009; 58(7): 1568 - 1577. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M B Nickdel, P Conigliaro, G Valesini, S Hutchison, R Benson, R V Bundick, A J Leishman, I B McInnes, J M Brewer, and P Garside Dissecting the contribution of innate and antigen-specific pathways to the breach of self-tolerance observed in a murine model of arthritis Ann Rheum Dis, June 1, 2009; 68(6): 1059 - 1066. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P.R. Sutherland, T. Van Belle, A. L. Wurster, A. Suto, M. Michaud, D. Zhang, M. J. Grusby, and M. von Herrath Interleukin-21 Is Required for the Development of Type 1 Diabetes in NOD Mice Diabetes, May 1, 2009; 58(5): 1144 - 1155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Henry, C. A. Acevedo-Suarez, and J. W. Thomas Functional Silencing Is Initiated and Maintained in Immature Anti-Insulin B Cells J. Immunol., March 15, 2009; 182(6): 3432 - 3439. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Zekavat, S. Y. Rostami, A. Badkerhanian, R. F. Parsons, B. Koeberlein, M. Yu, C. D. Ward, T.-S. Migone, L. Yu, G. S. Eisenbarth, et al. In Vivo BLyS/BAFF Neutralization Ameliorates Islet-Directed Autoimmunity in Nonobese Diabetic Mice J. Immunol., December 1, 2008; 181(11): 8133 - 8144. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-G. Chen, F. Scheuplein, M. A. Osborne, S.-W. Tsaih, H. D. Chapman, and D. V. Serreze Idd9/11 Genetic Locus Regulates Diabetogenic Activity of CD4 T-Cells in Nonobese Diabetic (NOD) Mice Diabetes, December 1, 2008; 57(12): 3273 - 3280. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Fiorina, A. Vergani, S. Dada, M. Jurewicz, M. Wong, K. Law, E. Wu, Z. Tian, R. Abdi, I. Guleria, et al. Targeting CD22 Reprograms B-Cells and Reverses Autoimmune Diabetes Diabetes, November 1, 2008; 57(11): 3013 - 3024. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Gavanescu, C. Benoist, and D. Mathis B cells are required for Aire-deficient mice to develop multi-organ autoinflammation: A therapeutic approach for APECED patients PNAS, September 2, 2008; 105(35): 13009 - 13014. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Galletti, C. Canones, P. Morande, M. Borge, P. Oppezzo, J. Geffner, R. Bezares, R. Gamberale, and M. Giordano Chronic Lymphocytic Leukemia Cells Bind and Present the Erythrocyte Protein Band 3: Possible Role as Initiators of Autoimmune Hemolytic Anemia J. Immunol., September 1, 2008; 181(5): 3674 - 3683. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Huang, I. J. Fugier-Vivier, T. Miller, M. J. Elliott, H. Xu, L. D. Bozulic, P. M. Chilton, and S. T. Ildstad Plasmacytoid Precursor Dendritic Cells From NOD Mice Exhibit Impaired Function: Are They a Component of Diabetes Pathogenesis? Diabetes, September 1, 2008; 57(9): 2360 - 2370. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yu, R. Dunn, M. R. Kehry, and H. Braley-Mullen B Cell Depletion Inhibits Spontaneous Autoimmune Thyroiditis in NOD.H-2h4 Mice J. Immunol., June 1, 2008; 180(11): 7706 - 7713. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Marleau, K. L. Summers, and B. Singh Differential Contributions of APC Subsets to T Cell Activation in Nonobese Diabetic Mice J. Immunol., April 15, 2008; 180(8): 5235 - 5249. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. M. Brodie, M. Wallberg, P. Santamaria, F. S. Wong, and E. A. Green B-Cells Promote Intra-Islet CD8+ Cytotoxic T-Cell Survival to Enhance Type 1 Diabetes Diabetes, April 1, 2008; 57(4): 909 - 917. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Xiu, C. P. Wong, J.-D. Bouaziz, Y. Hamaguchi, Y. Wang, S. M. Pop, R. M. Tisch, and T. F. Tedder B Lymphocyte Depletion by CD20 Monoclonal Antibody Prevents Diabetes in Nonobese Diabetic Mice despite Isotype-Specific Differences in Fc{gamma}R Effector Functions J. Immunol., March 1, 2008; 180(5): 2863 - 2875. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Marino, M. Batten, J. Groom, S. Walters, D. Liuwantara, F. Mackay, and S. T. Grey Marginal-Zone B-Cells of Nonobese Diabetic Mice Expand With Diabetes Onset, Invade the Pancreatic Lymph Nodes, and Present Autoantigen to Diabetogenic T-Cells Diabetes, February 1, 2008; 57(2): 395 - 404. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-D. Bouaziz, K. Yanaba, G. M. Venturi, Y. Wang, R. M. Tisch, J. C. Poe, and T. F. Tedder Therapeutic B cell depletion impairs adaptive and autoreactive CD4+ T cell activation in mice PNAS, December 26, 2007; 104(52): 20878 - 20883. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hussain and T. L. Delovitch Intravenous Transfusion of BCR-Activated B Cells Protects NOD Mice from Type 1 Diabetes in an IL-10-Dependent Manner J. Immunol., December 1, 2007; 179(11): 7225 - 7232. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Bour-Jordan, B. L. Salomon, H. L. Thompson, R. Santos, A. K. Abbas, and J. A. Bluestone Constitutive Expression of B7-1 on B Cells Uncovers Autoimmunity toward the B Cell Compartment in the Nonobese Diabetic Mouse J. Immunol., July 15, 2007; 179(2): 1004 - 1012. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Duddy, M. Niino, F. Adatia, S. Hebert, M. Freedman, H. Atkins, H. J. Kim, and A. Bar-Or Distinct Effector Cytokine Profiles of Memory and Naive Human B Cell Subsets and Implication in Multiple Sclerosis J. Immunol., May 15, 2007; 178(10): 6092 - 6099. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. J. Zunino, D. H. Storms, and C. B. Stephensen Diets Rich in Polyphenols and Vitamin A Inhibit the Development of Type I Autoimmune Diabetes in Nonobese Diabetic Mice J. Nutr., May 1, 2007; 137(5): 1216 - 1221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Puertas, J. Carrillo, X. Pastor, R. M. Ampudia, A. Alba, R. Planas, R. Pujol-Borrell, M. Vives-Pi, and J. Verdaguer Phenotype and Functional Characteristics of Islet-Infiltrating B-Cells Suggest the Existence of Immune Regulatory Mechanisms in Islet Milieu Diabetes, April 1, 2007; 56(4): 940 - 949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-G. Chen, P. A. Silveira, M. A. Osborne, H. D. Chapman, and D. V. Serreze Cellular Expression Requirements for Inhibition of Type 1 Diabetes by a Dominantly Protective Major Histocompatibility Complex Haplotype Diabetes, February 1, 2007; 56(2): 424 - 430. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Silveira, H. D. Chapman, J. Stolp, E. Johnson, S. L. Cox, K. Hunter, L. S. Wicker, and D. V. Serreze Genes within the Idd5 and Idd9/11 Diabetes Susceptibility Loci Affect the Pathogenic Activity of B Cells in Nonobese Diabetic Mice J. Immunol., November 15, 2006; 177(10): 7033 - 7041. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Rowe, T. Banovic, K. P. MacDonald, R. Kuns, A. L. Don, E. S. Morris, A. C. Burman, H. M. Bofinger, A. D. Clouston, and G. R. Hill Host B cells produce IL-10 following TBI and attenuate acute GVHD after allogeneic bone marrow transplantation Blood, October 1, 2006; 108(7): 2485 - 2492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. J. Quinn III, N. Noorchashm, J. E. Crowley, A. J. Reed, H. Noorchashm, A. Naji, and M. P. Cancro Cutting edge: impaired transitional B cell production and selection in the nonobese diabetic mouse. J. Immunol., June 15, 2006; 176(12): 7159 - 7164. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Hayward, N. Bautista-Lopez, K. Suzuki, A. Atrazhev, P. Dickie, and J. F. Elliott CD4 T Cells Play Major Effector Role and CD8 T Cells Initiating Role in Spontaneous Autoimmune Myocarditis of HLA-DQ8 Transgenic IAb Knockout Nonobese Diabetic Mice. J. Immunol., June 15, 2006; 176(12): 7715 - 7725. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Keir, S. C. Liang, I. Guleria, Y. E. Latchman, A. Qipo, L. A. Albacker, M. Koulmanda, G. J. Freeman, M. H. Sayegh, and A. H. Sharpe Tissue expression of PD-L1 mediates peripheral T cell tolerance J. Exp. Med., April 17, 2006; 203(4): 883 - 895. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yu, P. K. Maiti, M. Dyson, R. Jain, and H. Braley-Mullen B cell-deficient NOD.H-2h4 mice have CD4+CD25+ T regulatory cells that inhibit the development of spontaneous autoimmune thyroiditis J. Exp. Med., February 21, 2006; 203(2): 349 - 358. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Tian, D. Zekzer, Y. Lu, H. Dang, and D. L. Kaufman B Cells Are Crucial for Determinant Spreading of T Cell Autoimmunity among beta Cell Antigens in Diabetes-Prone Nonobese Diabetic Mice J. Immunol., February 15, 2006; 176(4): 2654 - 2661. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C.-H. Lee, P. C. Reifsnyder, J. K. Naggert, C. Wasserfall, M. A. Atkinson, J. Chen, and E. H. Leiter Novel Leptin Receptor Mutation in NOD/LtJ Mice Suppresses Type 1 Diabetes Progression: I. Pathophysiological Analysis Diabetes, September 1, 2005; 54(9): 2525 - 2532. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. J. Woodward and J. W. Thomas Multiple Germline {kappa} Light Chains Generate Anti-Insulin B Cells in Nonobese Diabetic Mice J. Immunol., July 15, 2005; 175(2): 1073 - 1079. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. Moore, H. Noorchashm, T. H. Lin, S. A. Greeley, and A. Naji NOD B-cells Are Insufficient to Incite T-Cell-Mediated Anti-islet Autoimmunity Diabetes, July 1, 2005; 54(7): 2019 - 2025. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Bezbradica, A. K. Stanic, N. Matsuki, H. Bour-Jordan, J. A. Bluestone, J. W. Thomas, D. Unutmaz, L. Van Kaer, and S. Joyce Distinct Roles of Dendritic Cells and B Cells in Va14Ja18 Natural T Cell Activation In Vivo J. Immunol., April 15, 2005; 174(8): 4696 - 4705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Rolf, V. Motta, N. Duarte, M. Lundholm, E. Berntman, M.-L. Bergman, L. Sorokin, S. L. Cardell, and D. Holmberg The Enlarged Population of Marginal Zone/CD1dhigh B Lymphocytes in Nonobese Diabetic Mice Maps to Diabetes Susceptibility Region Idd11 J. Immunol., April 15, 2005; 174(8): 4821 - 4827. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. O'Neill, M. J. Shlomchik, T. T. Glant, Y. Cao, P. D. Doodes, and A. Finnegan Antigen-Specific B Cells Are Required as APCs and Autoantibody-Producing Cells for Induction of Severe Autoimmune Arthritis J. Immunol., March 15, 2005; 174(6): 3781 - 3788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Hall, M. A. Vickers, E. McLeod, and R. N. Barker Rh autoantigen presentation to helper T cells in chronic lymphocytic leukemia by malignant B cells Blood, March 1, 2005; 105(5): 2007 - 2015. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hussain and T. L. Delovitch Dysregulated B7-1 and B7-2 Expression on Nonobese Diabetic Mouse B Cells Is Associated with Increased T Cell Costimulation and the Development of Insulitis J. Immunol., January 15, 2005; 174(2): 680 - 687. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Acevedo-Suarez, C. Hulbert, E. J. Woodward, and J. W. Thomas Uncoupling of Anergy from Developmental Arrest in Anti-Insulin B Cells Supports the Development of Autoimmune Diabetes J. Immunol., January 15, 2005; 174(2): 827 - 833. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Carrillo, M. C. Puertas, A. Alba, R. M. Ampudia, X. Pastor, R. Planas, N. Riutort, N. Alonso, R. Pujol-Borrell, P. Santamaria, et al. Islet-infiltrating B-Cells in Nonobese Diabetic Mice Predominantly Target Nervous System Elements Diabetes, January 1, 2005; 54(1): 69 - 77. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Alba, M. C. Puertas, J. Carrillo, R. Planas, R. Ampudia, X. Pastor, F. Bosch, R. Pujol-Borrell, J. Verdaguer, and M. Vives-Pi IFN{beta} Accelerates Autoimmune Type 1 Diabetes in Nonobese Diabetic Mice and Breaks the Tolerance to {beta} Cells in Nondiabetes-Prone Mice J. Immunol., December 1, 2004; 173(11): 6667 - 6675. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Salam, K. Matin, N. Matsumoto, Y. Tsuha, N. Hanada, and H. Senpuku E2f1 Mutation Induces Early Onset of Diabetes and Sjogren's Syndrome in Nonobese Diabetic Mice J. Immunol., October 15, 2004; 173(8): 4908 - 4918. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. S. Wong, L. Wen, M. Tang, M. Ramanathan, I. Visintin, J. Daugherty, L. G. Hannum, C. A. Janeway Jr, and M. J. Shlomchik Investigation of the Role of B-Cells in Type 1 Diabetes in the NOD Mouse Diabetes, October 1, 2004; 53(10): 2581 - 2587. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Matos, R. Park, D. Mathis, and C. Benoist Progression to Islet Destruction in a Cyclophosphamide-Induced Transgenic Model: A Microarray Overview Diabetes, September 1, 2004; 53(9): 2310 - 2321. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. H. Koonce and E. K. Bikoff Dissecting MHC Class II Export, B Cell Maturation, and DM Stability Defects in Invariant Chain Mutant Mice J. Immunol., September 1, 2004; 173(5): 3271 - 3280. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hussain, K. V. Salojin, and T. L. Delovitch Hyperresponsiveness, Resistance to B-Cell Receptor--Dependent Activation-Induced Cell Death, and Accumulation of Hyperactivated B-Cells in Islets Is Associated With the Onset of Insulitis but not Type 1 Diabetes Diabetes, August 1, 2004; 53(8): 2003 - 2011. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Havari, A. M. Lennon-Dumenil, L. Klein, D. Neely, J. A. Taylor, M. F. McInerney, K. W. Wucherpfennig, and M. A. Lipes Expression of the B7.1 Costimulatory Molecule on Pancreatic {beta} Cells Abrogates the Requirement for CD4 T Cells in the Development of Type 1 Diabetes J. Immunol., July 15, 2004; 173(2): 787 - 796. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Silveira, J. Dombrowsky, E. Johnson, H. D. Chapman, D. Nemazee, and D. V. Serreze B Cell Selection Defects Underlie the Development of Diabetogenic APCs in Nonobese Diabetic Mice J. Immunol., April 15, 2004; 172(8): 5086 - 5094. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Dromey, S. M. Weenink, G. H. Peters, J. Endl, P. J. Tighe, I. Todd, and M. R. Christie Mapping of Epitopes for Autoantibodies to the Type 1 Diabetes Autoantigen IA-2 by Peptide Phage Display and Molecular Modeling: Overlap of Antibody and T Cell Determinants J. Immunol., April 1, 2004; 172(7): 4084 - 4090. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. H. Schott, B. D. Haskell, H. M. Tse, M. J. Milton, J. D. Piganelli, C. M. Choisy-Rossi, P. C. Reifsnyder, A. V. Chervonsky, and E. H. Leiter Caspase-1 Is Not Required for Type 1 Diabetes in the NOD Mouse Diabetes, January 1, 2004; 53(1): 99 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Ugrinovic, N. Menager, N. Goh, and P. Mastroeni Characterization and Development of T-Cell Immune Responses in B-Cell-Deficient (Igh-6-/-) Mice with Salmonella enterica Serovar TyphimuriumInfection Infect. Immun., December 1, 2003; 71(12): 6808 - 6819. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Trembleau, G. Penna, S. Gregori, N. Giarratana, and L. Adorini IL-12 Administration Accelerates Autoimmune Diabetes in Both Wild-Type and IFN-{gamma}-Deficient Nonobese Diabetic Mice, Revealing Pathogenic and Protective Effects of IL-12-Induced IFN-{gamma} J. Immunol., June 1, 2003; 170(11): 5491 - 5501. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. T. Robles, G. S. Eisenbarth, N. J.M. Dailey, L. B. Peterson, and L. S. Wicker Insulin Autoantibodies Are Associated With Islet Inflammation But Not Always Related to Diabetes Progression in NOD Congenic Mice Diabetes, March 1, 2003; 52(3): 882 - 886. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. S. Sobel, M. Satoh, Y. Chen, E. K. Wakeland, and L. Morel The Major Murine Systemic Lupus Erythematosus Susceptibility Locus Sle1 Results in Abnormal Functions of Both B and T Cells J. Immunol., September 1, 2002; 169(5): 2694 - 2700. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. H. Nielsen and R. G. Q. Leslie Complement's participation in acquired immunity J. Leukoc. Biol., August 1, 2002; 72(2): 249 - 261. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Jaume, S. L. Parry, A.-M. Madec, G. Sonderstrup, and S. Baekkeskov Suppressive Effect of Glutamic Acid Decarboxylase 65-Specific Autoimmune B Lymphocytes on Processing of T Cell Determinants Located Within the Antibody Epitope J. Immunol., July 15, 2002; 169(2): 665 - 672. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kita, Z.-X. Lian, J. Van de Water, X.-S. He, S. Matsumura, M. Kaplan, V. Luketic, R. L. Coppel, A. A. Ansari, and M. E. Gershwin Identification of HLA-A2-restricted CD8+ Cytotoxic T Cell Responses in Primary Biliary Cirrhosis: T Cell Activation Is Augmented by Immune Complexes Cross-Presented by Dendritic Cells J. Exp. Med., January 7, 2002; 195(1): 113 - 123. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. P. L. Chiu, A. M. Jevnikar, and J. S. Danska Genetic Control of T and B Lymphocyte Activation in Nonobese Diabetic Mice J. Immunol., December 15, 2001; 167(12): 7169 - 7179. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Rivera, C.-C. Chen, N. Ron, J. P. Dougherty, and Y. Ron Role of B cells as antigen-presenting cells in vivo revisited: antigen-specific B cells are essential for T cell expansion in lymph nodes and for systemic T cell responses to low antigen concentrations Int. Immunol., December 1, 2001; 13(12): 1583 - 1593. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Yang, M. Bao, and J.-W. Yoon Intrinsic Defects in the T-Cell Lineage Results in Natural Killer T-Cell Deficiency and the Development of Diabetes in the Nonobese Diabetic Mouse Diabetes, December 1, 2001; 50(12): 2691 - 2699. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Hulbert, B. Riseili, M. Rojas, and J. W. Thomas Cutting Edge: B Cell Specificity Contributes to the Outcome of Diabetes in Nonobese Diabetic Mice J. Immunol., November 15, 2001; 167(10): 5535 - 5538. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. W. Greeley, D. J. Moore, H. Noorchashm, L. E. Noto, S. Y. Rostami, A. Schlachterman, H. K. Song, B. Koeberlein, C. F. Barker, and A. Naji Impaired Activation of Islet-Reactive CD4 T Cells in Pancreatic Lymph Nodes of B Cell-Deficient Nonobese Diabetic Mice J. Immunol., October 15, 2001; 167(8): 4351 - 4357. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Takemura, P. A. Klimiuk, A. Braun, J. J. Goronzy, and C. M. Weyand T Cell Activation in Rheumatoid Synovium Is B Cell Dependent J. Immunol., October 15, 2001; 167(8): 4710 - 4718. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Martin, D. Wolf-Eichbaum, G. Duinkerken, W. A. Scherbaum, H. Kolb, J. G. Noordzij, and B. O. Roep Development of Type 1 Diabetes despite Severe Hereditary B-Cell Deficiency N. Engl. J. Med., October 4, 2001; 345(14): 1036 - 1040. [Full Text] [PDF]
|
|
|
|
|
|
E. A. Johnson, P. Silveira, H. D. Chapman, E. H. Leiter, and D. V. Serreze Inhibition of Autoimmune Diabetes in Nonobese Diabetic Mice by Transgenic Restoration of H2-E MHC Class II Expression: Additive, But Unequal, Involvement of Multiple APC Subtypes J. Immunol., August 15, 2001; 167(4): 2404 - 2410. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Tian, D. Zekzer, L. Hanssen, Y. Lu, A. Olcott, and D. L. Kaufman Lipopolysaccharide-Activated B Cells Down-Regulate Th1 Immunity and Prevent Autoimmune Diabetes in Nonobese Diabetic Mice J. Immunol., July 15, 2001; 167(2): 1081 - 1089. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. Weaver Jr., B. Liu, and R. Tisch Plasmid DNAs Encoding Insulin and Glutamic Acid Decarboxylase 65 Have Distinct Effects on the Progression of Autoimmune Diabetes in Nonobese Diabetic Mice J. Immunol., July 1, 2001; 167(1): 586 - 592. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Abiru, A. K. Maniatis, L. Yu, D. Miao, H. Moriyama, D. Wegmann, and G. S. Eisenbarth Peptide and Major Histocompatibility Complex-Specific Breaking of Humoral Tolerance to Native Insulin With the B9-23 Peptide in Diabetes-Prone and Normal Mice Diabetes, June 1, 2001; 50(6): 1274 - 1281. [Abstract] [Full Text]
|
|
|
|
|
|
P. P.L. Chiu, D. V. Serreze, and J. S. Danska Development and Function of Diabetogenic T-cells in B-cell-Deficient Nonobese Diabetic Mice Diabetes, April 1, 2001; 50(4): 763 - 770. [Abstract] [Full Text]
|
|
|
|
|
|
H. Braley-Mullen and S. Yu Early Requirement for B Cells for Development of Spontaneous Autoimmune Thyroiditis in NOD.H-2h4 Mice J. Immunol., December 15, 2000; 165(12): 7262 - 7269. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.-J. Linton, J. Harbertson, and L. M. Bradley A Critical Role for B Cells in the Development of Memory CD4 Cells J. Immunol., November 15, 2000; 165(10): 5558 - 5565. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Holz, T. Dyrberg, W. Hagopian, D. Homann, M. v. Herrath, and M. B. A. Oldstone Neither B Lymphocytes Nor Antibodies Directed Against Self Antigens of the Islets of Langerhans Are Required for Development of Virus-Induced Autoimmune Diabetes J. Immunol., November 15, 2000; 165(10): 5945 - 5953. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Kolm-Litty, S. Berlo, E. Bonifacio, M. Bearzatto, A. M. Engel, M. Christie, A.-G. Ziegler, T. Wild, and J. Endl Human Monoclonal Antibodies Isolated from Type I Diabetes Patients Define Multiple Epitopes in the Protein Tyrosine Phosphatase-Like IA-2 Antigen J. Immunol., October 15, 2000; 165(8): 4676 - 4684. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Noorchashm, D. J. Moore, L. E. Noto, N. Noorchashm, A. J. Reed, A. L. Reed, H. K. Song, R. Mozaffari, A. M. Jevnikar, C. F. Barker, et al. Impaired CD4 T Cell Activation Due to Reliance Upon B Cell-Mediated Costimulation in Nonobese Diabetic (NOD) Mice J. Immunol., October 15, 2000; 165(8): 4685 - 4696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Myers, J. M. Davies, J. C. Tong, J. Whisstock, M. Scealy, I. R. Mackay, and M. J. Rowley Conformational Epitopes on the Diabetes Autoantigen GAD65 Identified by Peptide Phage Display and Molecular Modeling J. Immunol., October 1, 2000; 165(7): 3830 - 3838. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kondo, I. Iwata, K. Anzai, T. Akashi, S. Wakana, K. Ohkubo, H. Katsuta, J. Ono, T. Watanabe, Y. Niho, et al. Suppression of insulitis and diabetes in B cell-deficient mice treated with streptozocin: B cells are essential for the TCR clonotype spreading of islet-infiltrating T cells Int. Immunol., July 1, 2000; 12(7): 1075 - 1083. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. D. Shultz, P. A. Lang, S. W. Christianson, B. Gott, B. Lyons, S. Umeda, E. Leiter, R. Hesselton, E. J. Wagar, J. H. Leif, et al. NOD/LtSz-Rag1null Mice: An Immunodeficient and Radioresistant Model for Engraftment of Human Hematolymphoid Cells, HIV Infection, and Adoptive Transfer of NOD Mouse Diabetogenic T Cells J. Immunol., March 1, 2000; 164(5): 2496 - 2507. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. Green, F. S. Wong, K. Eshima, C. Mora, and R. A. Flavell Neonatal Tumor Necrosis Factor {alpha} Promotes Diabetes in Nonobese Diabetic Mice by Cd154-Independent Antigen Presentation to Cd8+ T Cells J. Exp. Med., January 17, 2000; 191(2): 225 - 238. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Kukreja Autoimmunity and Diabetes J. Clin. Endocrinol. Metab., December 1, 1999; 84(12): 4371 - 4378. [Abstract] [Full Text]
|
|
|
|
|
|
M. E. Ozaki, B. A. Coren, T. N. Huynh, D. J. Redondo, H. Kikutani, and S. R. Webb CD4+ T Cell Responses to CD40-Deficient APCs: Defects in Proliferation and Negative Selection Apply Only with B Cells as APCs J. Immunol., November 15, 1999; 163(10): 5250 - 5256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Noorchashm, Y. K. Lieu, N. Noorchashm, S. Y. Rostami, S. A. S. Greeley, A. Schlachterman, H. K. Song, L. E. Noto, A. M. Jevnikar, C. F. Barker, et al. I-Ag7-Mediated Antigen Presentation by B Lymphocytes Is Critical in Overcoming a Checkpoint in T Cell Tolerance to Islet {beta} Cells of Nonobese Diabetic Mice J. Immunol., July 15, 1999; 163(2): 743 - 750. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
