By
From the Department of Pathology and Center for Immunology, Washington University School of Medicine, St. Louis, Missouri 63110
Autoimmune diabetes is caused by the CD4+, T helper 1 (Th1) cell-mediated apoptosis of insulin-producing cells. We have previously shown that Th2 T cells bearing the same T cell
receptor (TCR) as the diabetogenic Th1 T cells invade islets in neonatal nonobese diabetic
(NOD) mice but fail to cause disease. Moreover, when mixed in excess and cotransferred with
Th1 T cells, Th2 T cells could not protect NOD neonates from Th1-mediated diabetes. We have now found, to our great surprise, the same Th2 T cells that produced a harmless insulitis
in neonatal NOD mice produced intense and generalized pancreatitis and insulitis associated
with islet cell necrosis, abscess formation, and subsequent diabetes when transferred into immunocompromised NOD.scid mice. These lesions resembled allergic inflamation and contained a
large eosinophilic infiltrate. Moreover, the Th2-mediated destruction of islet cells was mediated by local interleukin-10 (IL-10) production but not by IL-4. These findings indicate that
under certain conditions Th2 T cells may not produce a benign or protective insulitis but rather acute pathology and disease. Additionally, these results lead us to question the feasibility
of Th2-based therapy in type I diabetes, especially in immunosuppressed recipients of islet cell
transplants.
Insulin-dependent diabetes mellitus (IDDM)1 is caused by
the autoimmune destruction of insulin-producing CD4+ T cells can be differentiated into at least two major subsets, Th1 cells that secrete IFN- We have previously reported that the ability of CD4+
T cells to transfer diabetes to naïve recipients resided not
with the antigen specificity recognized by the TCR, per se,
but with the phenotypic nature of the T cell response (12).
Strongly polarized Th1 T cells transferred disease into NOD
neonatal mice, while Th2 T cells did not, despite being activated and bearing the same TCR as the diabetogenic Th1
T cell population. Moreover, upon cotransfer, Th2 T cells
could not ameliorate the Th1-induced diabetes, even when
Th2 cells were cotransferred in ~10-fold excess (12).
However, we wondered if we transferred Th2 cells before transfer of Th1 cells, we might amplify the protective
influence of Th2 cells, and thereby control the subsequent
Th1 T cell response. In this study, we attempt to evaluate
directly this hypothesis by producing Th1 and Th2 T cells
from the islet-reactive BDC2.5 TCR transgenic NOD mouse
(20), and then transfer sequentially Th2 and then Th1 T cells
into NOD recipients mice.
Much to our surprise, we found that Th2 T cells exerted
markedly differing effects on NOD recipients depending
on the immune status of the recipient mouse. Here, we report that while Th1-polarized T cells can transfer disease in
neonatal NOD mice, something Th2-polarized T cell fail
to do, both Th1- and Th2-polarized T cells can transfer disease in NOD.scid mice and other immune-compromised
recipients. The Th2-mediated diabetes in NOD.scid recipients exhibited a longer prediabetic phase and a lowered overall incidence. Moreover, the diabetic lesion created by Th2
cells was unique and quite unlike the lesion found in spontaneously diabetic BDC2.5 mice or Th1 T cell-induced diabetes in either neonates or NOD.scid mice. That Th2 cells
caused a distinct but important disease in immune-compromised recipients has significant implications with regard to
the potential use of Th2 cells as a therapeutic agent.
Mice.
BDC2.5 TCR transgenic mice were described previously (20). Mice used in these experiments were housed under
specific pathogen-free conditions and were backcrossed to NOD/lt
for >20 generations. The NOD.scid mice were bred under
pathogen-free conditions at Washington University from an original breeding stock provided by Dr. E. Leiter of the Jackson Laboratories.
Flow Cytometry.
Flow cytometry was performed on either a
Becton Dickinson FACScan® or FACSVantage® flow cytometer.
We purchased anti-CD4-PE mAb (Caltag Laboratory, South San
Francisco, CA). mAb to the Diabetes.
Diabetes was assessed by measurement of venous
blood using a Bayer Glucometer Elite one-step blood glucose meter.
Animals were considered diabetic after two consecutive measurements Immunohistochemistry.
Mice were killed by cervical dislocation
or CO2 asphyxiation. The entire pancreata were removed, fixed
either in 10% neutral-buffered formalin at 4°C for at least 20 h
but not more than 26 h, embedded in paraffin, and sections (2 µm),
collected on poly L-lysine coated slides (VWR Scientific Products, Corp., Philadelphia, PA). Alternatively, pancreata were snap
frozen in OCT compound (Tissue-tek) for cryosectioning. 5-µm
cryosections were obtained, air dried, and stored at cells
in the islets of Langerhans of the pancreas (1, 2). The leukocytic infiltration, termed insulitis, is a heterogeneous mixture
of CD4+ and CD8+ T lymphocytes, B lymphocytes, macrophage and dendritic cells (3). In general, T lymphocytes,
play the most pivotal role in initiating the disease process
(4). Recently, we demonstrated that islet cell antigen-responsive CD4+ T cells alone are sufficient for the spontaneous development and transfer of diabetes in NOD.scid
mice (8).
and Th2 cells that
produce IL-4 and IL-10. Th1 cells are critically involved in
the generation of effective cellular immunity (9), whereas
Th2 T cells are instrumental in the generation of humoral
and mucosal immunity and allergy, including the activation
of eosinophils and mast cells and the production of IgE (9).
A number of studies have now correlated diabetes with
Th1 phenotype development (10). On the other hand,
Th2 T cells were shown to be relatively innocuous (12-
15). Some have even speculated that Th2 T cells in fact, may be protective (16), although direct evidence in
support of Th2-mediated protection is lacking.
chain of the transgenic TCR,
KT4-10 (21), was purified from ascites and conjugated with FITC. List mode data was collected on 1 × 105 cells and reanalyzed using WinMDI (version 2.1.4) software written by J. Trotter (http://facs.scripps.edu).
250 mg/dl (13.75 mM). Onset of diabetes was dated
from the first consecutive reading. In most instances, sustained
hyperglycemia of >500 mg/dl was observed, and animals were
killed to avoid prolonged discomfort.
20°C until
used. Formalin-fixed sections were deparaffinized in xylene and
alcohol, and stained with hematoxylin and eosin for general morphology.
T Cell Transfers.
Naive CD4+ T cells were enriched from
thymi of BDC2.5/NOD mice by depleting CD4+ CD8+ double-positive cells with anti-CD8 antibody and rabbit complement (Cederlane, Ontario, Canada). T cells were resuspended at 1 × 105 cells/ml in DMEM supplemented with 10% heat-inactivated
fetal bovine serum, 1 mM sodium pyruvate, 1 mM glutamine,
and 50 µM 2-mercaptoethanol and incubated with 2 × 106
APC/ml and 2 × 104 islet cells in conditions previously demonstrated to generate Th1 or Th2 cells for 7 d (12, 23). In brief,
T cells were cultured in the presence of islet cells and NOD splenocytes in the presence of IFN- (50 U/ml) and IL-12 (50 U/ml),
and neutralizing anti-IL-4 antibodies (11B11, 10 µg/ml) for generation of Th1 T cells or IL-4 (50 U/ml) and neutralizing amounts
of anti IFN-
(H22, 10 µg/ml) and anti IL-12 (Tosh, 10 µg/ml)
antibodies for Th2 T cells. T cells were recovered from these cultures and transferred into neonatal NOD or 6-wk-old NOD.scid
mice. An aliquot of cells was restimulated with antigen and APC
alone and supernatants were checked for the presence of appropriate cytokines by ELISA to confirm polarization of T cells.
Neonatal NOD mice were injected with 1 × 106 CD4+ V
4+
Th1 or Th2 T cells intraperitoneally, whereas NOD.scid and
TCR-
-deficient NOD (NOD.C
/
) mice were injected
with 1 × 106 cells intravenously.
In Vitro Restimulation.
Th2 T cells stimulated as above were
recovered and cultured for an additional 7 d in the presence of
antigen under either Th2 conditions (anti-IL-12, anti-IFN-,
and IL-4), Th1 conditions (anti-IL-4, IFN-
, and IL-12), or
Th1/Th2 conditions (IL-4, IL-12, and IFN-
) to evaluate Th2
phenotype stablity. At the end of the second 7-d cycle, recovered
T cells were restimulated with islet cells and irradiated NOD
APC in the absence of exogenous cytokines or mAb, and supernatant were collected and analyzed by ELISA at 24 h.
Cytokine ELISA.
IL-4 was detected using monoclonal anti-IL-4 antibody (11B11) as the capture antibody and revealed with
rabbit anti-mouse IL-4 polyclonal serum (a gift of Dr. R.D.
Schreiber, Washington University, St. Louis, MO) followed by
biotin-conjugated goat anti-rabbit antibody (Vector) and Streptavidin-horseradish peroxidase (Jackson Immunoresearch, Avondale, PA) using TMB substrate (Sigma). IFN- was detected using H22 (gift of Dr. R.D. Schreiber) as the capture antibody and
revealed with goat anti-mIFN-
antibody followed by horseradish peroxidase-conjugated donkey anti-goat IgG (Jackson Immunoresearch) using ABTS (Sigma) as substrate.
We generated Th1 and Th2 cells from naive populations of BDC2.5/
NOD.scid thymocytes by in vitro stimulation with islet cells in the presence of anti-IL-4, recombinant IFN-, and IL-12
for Th1, or anti-IL-12 and anti-IFN-
, and recombinant
IL-4 for Th2 cells. T cells taken from these cultures were
restimulated in the presence of islet cells and APC alone and
the supernatants were tested for the presence of appropriate
signature cytokines by ELISA confirming the polarization
of the T cells (data not shown). Th1- or Th2-polarized
BDC2.5 T cells were then injected into either neonatal
NOD (immune-competent) mice or NOD.scid (immune-compromised) mice, and the recipient animals were monitored for diabetes by blood glucose. Th1-injected neonatal
NOD mice became diabetic by day 14, whereas Th2-injected
neonatal recipient mice remained normal glycemic even after 35 d (Fig. 1 a). Moreover, when Th1 T cells from the same
polarized cultures were transferred into 6-wk-old NOD.scid
mice, diabetes was likewise rapidly induced (Fig. 1 b). The
rate, progression, and penetrance of disease was identical to
that seen in neonatal recipients of Th1 cells (Fig. 1, a and
b), with sustained hyperglycemia appearing between day 9 and 14 after-transfer (Fig. 1 b). Insulitis, as determined from serial pancreas histology, started at day 5 and becoming severe by day 9 (data not shown). Moreover, when Th1 cells
were transferred into NOD mice lacking
-T cells
(NOD C
/
), hyperglycemia resulted with similar kinetics and penetrance (data not shown). Therefore, Th1 cells
produced similar disease in both immune-compromised
and immune-competent mice. In addition, the Th1-mediated disease was nearly identical to the diabetes seen with
BDC2.5/NOD.scid T cells from spontaneously diabetic animals (8).
Surprisingly, unlike the Th2 recipient neonatal NOD,
which remained disease free, immune-compromised NOD
mice (both NOD.scid and NOD C/
) infused with Th2
cells developed hyperglycemia (Fig. 1 b; data not shown).
However, the kinetics and penetrance of disease were different from that seen in Th1-mediated disease. Islet-reactive Th2 cells produced disease with slower kinetics and
only partial penetrance in NOD.scid recipients (Fig. 1 b).
By day 16, 25% of the mice were diabetic and maximum
disease penetrance was not achieved until day 24 with 60%
of the mice showing sustained hyperglycemia. This slow
disease seen in NOD.scid recipients of Th2 cells could be
due to (a) the slow expansion of a contaminating Th1 T cell population or a switch of the input T cell population to Th1
or (b) a unique Th2 lesion with inherently slower kinetics.
We addressed the first possibility by taking polarized Th2
T cells and assessing their ability to switch phenotypes both
in vitro and in vivo. In culture, naive BDC2.5 T cells were
stimulated under conditions that generated Th2 T cells as
above. However, after one round of culture, Th2 T cells
were then subjected to a secondary antigenic stimulation in
the presence of IL-12 and IFN- (Th1 condition) that favors the outgrowth of Th1 cells or the phenotypic switch of
poorly polarized Th2 T cells. The presence of IFN-
-producing cells is then revealed under neutral reconditions. Even
after 1 wk of culture with IFN-
and IL-12, we were unable to detect IFN-
production by our Th2 cells (data not
shown). More directly, splenocytes recovered from NOD.scid
recipient mice were stimulated with plate-bound anti-CD3,
and the supernatants were assayed for both Th1 and Th2
cytokines. We found that the recovered splenocytes could
only secrete IL-4 in detectable quantities (data not shown).
From this, we concluded that Th2 cells did not switch
phenotype after polarization, nor did a contaminating Th1 population grow out from the input Th2 transfer population.
The best evidence supporting a unique Th2-mediated disease was radically different histopathology of the Th2 lesions in NOD.scid mice. Th1 T cells and those from spontaneously diabetic
BDC2.5/NOD.scid mice produce focused islet lesions without involvement of the surrounding exocrine tissue (Fig. 2
a; reference 8). That is to say, the infiltrating mass of leucocytes surrounded the islets (peri-insulitis) and then proceeded to invade the islet from the periphery to the center
(insulitis). The infiltrating mass was predominantly V4+,
CD4+ T cells, macrophages with a few neutrophils (see below). By contrast, Th2 T cells followed a more disorganized pattern. Leucocytes infiltrated not only the islets but
also the exocrine tissue (Fig. 2 b) with significant damage
produced to the exocrine tissue as well as islets (Fig. 2, b
and f ). The Th2 lesions contained a swarming pancreatitis
predominantly composed of eosinophils and PMN cells (Fig. 2 f ). Additionally, the Th2 lesions were characterized by the presence of abscesses as evidenced by the massive accumulation of eosinophils and PMNs with necrotic cellular
debris with in the islets (Fig. 2, b and f ).
The organized outside-in pattern of Th1 T cell-mediated infiltration was associated with cell apoptosis. Insulin-containing
cells were present in the intact areas of
infiltrated islets (Fig. 2 c), whereas infiltrated areas saw a loss
of insulin-bearing islet cells. Th1 lesions showed
cells apoptosis (Fig. 2 e) similar to that seen in spontaneous disease
(8). There was a strict correlation between the presence of
insulitis and and
cell apoptosis. Cell death in the Th2 cell
infiltrated islets by contrast was due largely to necrosis and
subsequent abscess formation (Fig. 2 e). It is worth noting
that the islet cell necrosis in Th2 lesions was often asymmetric or segmental especially with large islets where one
portion of the islet would be completely necrotic and the
adjacent islet mass completely intact and producing insulin (Fig. 2 d).
Dramatic differences in the organization and cellular
makeup of the two lesions was further underscored upon
immunohistochemical labeling of pancreatic frozen sections
from NOD.scid mice infused with Th1 and Th2 T cells.
Fig. 3 a depicts a section of a Th1 T cell-induced lesion,
showing an accumulation of transgenic V4+ T cells peripheral to the infiltrated islet. The exocrine tissue was
largely devoid of any T cells. By contrast, in Th2 lesions, abscess formation was seen with a few weakly staining
CD4+, V
4+ T cells associated with both islet and the peripheral exocrine tissue (Fig. 3 b). In general, T cell staining
was weak and diffuse and made up only a small portion of
the total leukocytic infiltration within the inflamed pancreas. Macrophage subpopulations were present in both
Th1 and Th2 lesion as ascertained by CD11b, MOMA-1,
MOMA-2, and ER-MP23 staining (data not shown).
However, the Th2 lesions had a larger number of scavenger macrophages surrounding the islet abscesses.
Peripheral Node Addressin Is Not Expressed in the High Endothelial Venules of Th2 Recipient Mice.
Both Th1 and Th2
lesions showed mad-CAM on the high endothelial venules
with in the pancreas (Fig. 3, c-d). However, Th2 lesions lacked expression of the activated form, peripheral node
addressin, PNA, as detected by the MECA-79 mAb (Fig. 3,
e-f ). This was significant because PNA expression requires
localized production of IFN- and TNF-
, again suggesting that Th2 lesions lack significant production of these
Th1 cytokines.
To rule out the contribution of
neutrophils and granulocytes in the pancreatitis associated
with Th2 T cell transfer, we treated mice with a granulocyte-specific mAb RB6 at concentrations sufficient to severely deplete granulocytes in vivo (24, 25). We found no
change in disease or in the characteristics of the Th2 lesion when granulocytes and neutrophils were depleted from
mice injected with Th2 cells (data not shown). However,
when we treated Th2 recipient NOD.scid mice with neutralizing antibodies against IL-10, a Th2 cytokine previously
shown to have a major effect on autoimmune diabetes, the
Th2-mediated disease was significantly ameliorated. This
was not true for another Th2 cytokine, IL-4 (Fig. 4).
NOD.scid mice received either 250 µg of anti-IL-4, anti-
IL-10, or saline every 72 h, with the first dose 24 h before
infusion of the Th2 T cells. As depicted in Fig. 4, anti-IL-4
treatment did not protect from diabetes, whereas anti-IL-10
significantly delayed or abrogated the Th2 pathology. Histology performed on pancreas from these mice indicate no
change in the lesion characteristics, yet anti-IL-10 treatment substantially protected against Th2 lesions, suggesting that IL-10 contributes to the pathology of Th2 disease.
We reported that Th2 cells were capable of inducing hyperglycemia in immune-compromised NOD.scid mice but
not in neonatal NOD mice. The kinetics of disease was
slower when compared with Th1-mediated disease due to
a different mechanism of islet cell destruction. The Th2 lesions were characterized by a predominantly eosinophilic
infiltration, islet necrosis and abscess, and a severe pancreatitis with destruction of both exo- and endocrine tissue. By contrast, Th1 T cells produced focally confined infiltration of the islets and cell apoptosis which largely spares
the adjacent exocrine tissue. This was more reminiscent of
the lesions seen in the natural disease and in our spontaneous TCR transgenic model of disease (8, 12).
Two important question arise from these findings. First, how do Th2 T cells propagate the pancreatitis and necrosis of islet cells in the immune-compromised recipient? And, second, why is the Th2 lesion found exclusively in immune-compromised hosts? Addressing the former, our data suggest that a particularly important mediator of islet cell necrosis is IL-10. Interestingly, previously published reports using transgenic mice that produced IL-4 in the islets were protected from diabetes, whereas local production of IL-10 produced a severe disease (26). These observations agree well with our findings that anti-IL-10 treatment of Th2 T cell recipients greatly diminishes disease onset and concurrent pathology. Wogensen et al. (26) have suggested that localized production of IL-10 produces important changes in the vascular endothelium, which may lead to accumulation of T cells, macrophage and eosinophils by changing the vascular addressin expressed on the endothelium. However, if the local production of IL-10 by our Th2 T cells acted predominantly to stimulate homing, treatment of the recipients with anti-IL-10 should have diminished greatly the eosinophilic nature of infiltration. However, we found that anti-IL-10 had a far greater effect on the pathology of the lesion than on its cellularity. One hypothesis is that IL-10 leads to vascular damage resulting in hypoxia and subsequent abscess formation. Moritani et al. (28) have previously described ductal proliferation as an effect mediated by transgenic expression of IL-10. Alternatively, owing to the venation of microcapillaries that feed islets (29), localized endothelial damage produced by IL-10 could lead to vascular occlusion of one of the branches feeding the microvascular circulation of the islet, resulting in the segmented necrosis that affected many of the larger islets. However, we have not found any direct evidence to support this interpretation.
The susceptibility of immune-compromised animals to
Th2-mediated disease may lie with the activated state of
the innate immune system in the absence of specific immunity. Specifically, IL-10 secretion has pronounced effects
on mononuclear phagocyte development and can enhance
the development of the myelomonocytic lineage (30).
However, the most likely explanation rests with the lack of
additional -T cells. It is likely that Th2 cell regulation requires interactions with other
-T cells. Because in
both our results and those of Lafaille et al. (33), the exaggerated and deleterious Th2 phenotype occurs only in the
absence of diverse
-T cell compartment, how the presence of other T cells affect the in vivo function of Th2 remains to be seen.
It would be of obvious clinical advantage if one could deviate an ongoing inflammatory immune response to a benign Th2 response. Some have even suggested that Th2 T cells are suppressive or tolerant T cells, reviewed and discussed in references 11, 34. But it is clear that some doubts have existed as to the efficacious use of Th2 therapy to treat inflammatory-based diseases (35). Our results demonstrate that Th2 cells may not be all that benign, especially if pushed to extremes, and carry with them a real potential to cause clinical disease. This is of particular concern in the case at hand, autoimmune diabetes, where immune deviation therapy as proposed in conjunction with islet cell transplantation and immunosuppression may cause a severe necrotic destruction of engrafted islets. Moreover, this may not be a limited phenomenon as similar studies in experimental autoimmune encephalomyelitis yield remarkably similar results (33).
Address correspondence to J.D. Katz, Department of Pathology and Center for Immunology, Washington University School of Medicine, Campus Box 8118, 660 South Euclid, St. Louis, Missouri 63110. Phone: 314-747-1221; Fax: 314-747-0728; E-mail: jkatz{at}immunology.wustl.edu
Received for publication 25 April 1997 and in revised form 9 May 1997.
1 Abbreviations used in this paper: AEC, aminoethylcarbazole; DAB, diaminobenzidine; IDDM, insulin-dependent diabetes mellitus; PNA, peripheral node addressin.We wish to thank Dr. P. Lacy for critical review of this manuscript and helpful discussion on islet histopathology. We wish to thank Drs. R.D. Schreiber and E.R. Unanue and L. Lainer for gift of antibodies. We wish to thank Ms. O. Strots and M.L. Chivetta for excellent technical assistance.
This work is supported by the generous start-up funding of the Department of Pathology and a grant from the US Public Health Service and Juvenile Diabetes Foundation International 1 P01 AI/DK 39676. J.D. Katz is a recipient of a career development award of the American Diabetes Association.
1. | Bach, J.F.. 1991. Insulin-dependent diabetes mellitus. Curr. Opin. Immunol. 3: 902-905 [Medline]. |
2. |
Atkinson, M.A., and
N.K. Maclaren.
1994.
The pathogenesis
of insulin-dependent diabetes mellitus.
N. Engl. J. Med.
331:
1428-1436
|
3. | O'Reilly, L.A., P.R. Hutchings, P.R. Crocker, E. Simpson, T. Lund, D. Kiossis, F. Takei, J. Baird, and A. Cooke. 1991. Characterization of pancreatic islet cell infiltrates in NOD mice: effect of cell transfer and transgene expression. Eur. J. Immunol. 21: 1171-1180 [Medline]. |
4. | Sempe, P., P. Bedossa, M.F. Richard, M.C. Villa, J.F. Bach, and C. Boitard. 1991. .Anti-alpha/beta T cell receptor monoclonal antibody provides an efficient therapy for autoimmune diabetes in nonobese diabetic (NOD) mice. Eur. J. Immunol. 21: 1163-1169 [Medline]. |
5. | Miyazaki, A., T. Hanafusa, K. Yamada, J. Miyagawa, H. Fujino-Kurihara, H. Nakajima, K. Nonaka, and S. Tarui. 1985. Predominance of T lymphocytes in pancreatic islets and spleen of pre-diabetic non-obese diabetic (NOD) mice: a longitudinal study. Clin. Exp. Immunol. 60: 622-630 [Medline]. |
6. | Harada, M., and S. Makino. 1986. Suppression of overt diabetes in NOD mice by anti-thymocyte serum or anti-Thy 1, 2 antibody. Jikken. Dobutsu. 35: 501-504 [Medline]. |
7. | Makino, S., M. Harada, Y. Kishimoto, and Y. Hayashi. 1986. Absence of insulitis and overt diabetes in athymic nude mice with NOD genetic background. Jikken. Dobutsu. 35: 495-498 [Medline]. |
8. |
Kurrer, M.O.,
S.V. Pakala,
H.L. Hanson, and
J.D. Katz.
1997.
![]() |
9. | Abbas, A.K., K.M. Murphy, and A. Sher. 1996. Functional diversity of helper T lymphocytes. Nature (Lond.). 383: 787-793 [Medline]. |
10. | Healey, D., P. Ozegbe, S. Arden, P. Chandler, J. Hutton, and A. Cooke. 1995. In vivo activity and in vitro specificity of CD4+ Th1 and Th2 cells derived from the spleens of diabetic NOD mice. J. Clin. Invest. 95: 2979-2985 [Medline]. |
11. | Liblau, R.S., S.M. Singer, and H.O. McDevitt. 1995. Th1 and Th2 CD4+ T cells in the pathogenesis of organ-specific autoimmune diseases. Immunol. Today. 16: 34-38 [Medline]. |
12. | Katz, J.D., C. Benoist, and D. Mathis. 1995. T helper cell subsets in insulin-dependent diabetes. Science (Wash. DC). 268: 1185-1188 [Medline]. |
13. | Sarvetnick, N., J. Shizuru, D. Liggitt, L. Martin, B. McIntyre, A. Gregory, T. Parslow, and T. Stewart. 1990. Loss of pancreatic islet tolerance induced by beta-cell expression of interferon-gamma. Nature (Lond.). 346: 844-847 [Medline]. |
14. | Campbell, I.L., T.W. Kay, L. Oxbrow, and L.C. Harrison. 1991. Essential role for interferon-gamma and interleukin-6 in autoimmune insulin-dependent diabetes in NOD/Wehi mice. J. Clin. Invest. 87: 739-742 [Medline]. |
15. | Rapoport, M.J., A. Jaramillo, D. Zipris, A.H. Lazarus, D.V. Serreze, E.H. Leiter, P. Cyopick, J.S. Danska, and T.L. Delovitch. 1993. Interleukin 4 reverses T cell proliferative unresponsiveness and prevents the onset of diabetes in nonobese diabetic mice. J. Exp. Med. 178: 87-99 [Abstract]. |
16. | Shimada, A., B. Charlton, P. Rohane, C. Taylor-Edwards, and C.G. Fathman. 1996. Immune regulation in type 1 diabetes. J. Autoimmun. 9: 263-269 [Medline]. |
17. | Tian, J., M.A. Atkinson, M. Clare-Salzler, A. Herschenfeld, T. Forsthuber, P.V. Lehmann, and D.L. Laufman. 1996. Nasal administration of glutamate decarboxylase (GAD65) peptides induces Th2 responses and prevents murine insulin-dependent diabetes. J. Exp. Med. 183: 1561-1567 [Abstract]. |
18. | Rabinovitch, A.. 1994. Immunoregulatory and cytokine imbalances in the pathogenesis of IDDM. Therapeutic intervention by immunostimulation? Diabetes. 43: 613-621 [Abstract]. |
19. | Wogensen, L., L. Molony, D. Gu, T. Krahl, S. Zhu, and N. Sarvetnick. 1994. Postnatal anti-interferon-gamma treatment prevents pancreatic inflammation in transgenic mice with beta-cell expression of interferon-gamma. J. Interferon. Res. 14: 111-116 [Medline]. |
20. | Katz, J.D., B. Wang, K. Haskins, C. Benoist, and D. Mathis. 1993. Following a diabetogenic T cell from genesis through pathogenesis. Cell. 74: 1089-1100 [Medline]. |
21. | Haskins, K., M. Portas, B. Bergman, K. Lafferty, and B. Bradley. 1989. Pancreatic islet-specific T-cell clones from nonobese diabetic mice. Proc. Natl. Acad. Sci. USA. 86: 8000-8004 [Abstract]. |
22. |
Ben-Sasson, S.A.,
Y. Sherman, and
Y. Gavrieli.
1995.
Identification of dying cells![]() |
23. | Hsieh, C.S., S.E. Macatonia, C.S. Tripp, S.F. Wolf, A. O'Garra, and K.M. Murphy. 1993. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science (Wash. DC). 260: 547-549 [Medline]. |
24. |
Czuprynski, C.J.,
J.F. Brown,
N. Maroushek,
R.D. Wagner, and
H. Steinberg.
1994.
Administration of anti-granulocyte
mAb RB6-8C5 impairs the resistance of mice to Listeria monocytogenes infection.
J. Immunol.
152:
1836-1846
|
25. | Conlan, J.W., and R.J. North. 1994. Neutrophils are essential for early anti-Listeria defense in the liver, but not in the spleen or peritoneal cavity, as revealed by a granulocyte-depleting monoclonal antibody. J. Exp. Med. 179: 259-268 [Abstract]. |
26. | Wogensen, L., M.S. Lee, and N. Sarvetnick. 1994. Production of interleukin 10 by islet cells accelerates immune-mediated destruction of beta cells in nonobese diabetic mice. J. Exp. Med. 179: 1379-1384 [Abstract]. |
27. | Lee, M.S., L. Wogensen, J. Shizuru, M.B. Oldstone, and N. Sarvetnick. 1994. Pancreatic islet production of murine interleukin-10 does not inhibit immune-mediated tissue destruction. J. Clin. Invest. 93: 1332-1338 [Medline]. |
28. | Moritani, M., K. Yoshimoto, F. Tashiro, C. Hashimoto, J. Miyazaki, S. Ii, E. Kudo, H. Iwahana, Y. Hayashi, T. Sano, and M. Itakura. 1994. Transgenic expression of IL-10 in pancreatic islet A cells accelerates autoimmune insulitis and diabetes in non-obese diabetic mice. Int. Immunol. 6: 1927-1936 [Abstract]. |
29. | Brunicardi, F.C., J. Stagner, S. Bonner-Weir, H. Wayland, R. Kleinman, E. Livingston, P. Guth, M. Menger, R. McCuskey, M. Intaglietta, et al . 1996. Microcirculation of the islets of Langerhans. Diabetes. 45: 385-392 [Medline]. |
30. | Colotta, F., M. Sironi, A. Borre, W. Luini, F. Maddalena, and A. Mantovani. 1992. Interleukin 4 amplifies monocyte chemotactic protein and interleukin 6 production by endothelial cells. Cytokine. 4: 24-28 [Medline]. |
31. | Sironi, M., C. Munoz, T. Pollicino, A. Siboni, F.L. Sciacca, S. Bernasconi, A. Vecchi, F. Colotta, and A. Mantovani. 1993. Divergent effects of interleukin-10 on cytokine production by mononuclear phagocytes and endothelial cells. Eur. J. Immunol. 23: 2692-2695 [Medline]. |
32. | Calzada-Wack, J.C., M. Frankenberger, and H.W. Ziegler-Heitbrock. 1996. Interleukin-10 drives human monocytes to CD16 positive macrophages. J. Inflammation. 46: 78-85 . [Medline] |
33. |
Lafaille, J.J.,
F. Van de Keere,
A. Hsu,
J.L. Baron,
C.S. Raine, and
S. Tonegawa.
1997.
Mylein basic protein-specific Th2
cells cause experimental autoimmune encephalomyelitis in
immunodeficient hosts rather than protect them from the disease.
J. Exp. Med.
186:
299-306
|
34. | Constant, S.L., and K. Bottomly. 1997. Induction of Th1 and Th2 CD4+ T cell responses: the alterative approaches. Annu. Rev. Immunol. 15: 297-322 [Medline]. |
35. | Khoruts, A., S.D. Miller, and M.K. Jenkins. 1995. Neuroantigen-specific Th2 cells are inefficient suppressors of experimental autoimmune encephalomyelitis induced by effector Th1 cells. J. Immunol. 155: 5011-5017 [Abstract]. |
36. |
McFarland, H.F..
1996.
Complexities in the treatment of autoimmune disease.
Science (Wash. DC).
274:
2037-2038
|
37. |
Genain, C.P.,
K. Abel,
N. Belmar,
F. Villinger,
D.P. Rosenberg,
C. Linington,
C.S. Raine, and
S.L. Hauser.
1996.
Late
complications of immune deviation therapy in a nonhuman
primate.
Science (Wash. DC).
274:
2054-2057
|