bluuebunny/crossref_metadata_2025 huggingface dataset:
Tissue factor-factor VIIa–specific up-regulation of IL-8 expression in MDA-MB-231 cells is mediated by PAR-2 and results in increased cell migration
Authors: Hjortoe Gertrud M. , Petersen Lars C. , Albrektsen Tatjana , Sorensen Brit B. , Norby Peder L. , Mandal Samir K. , Pendurthi Usha R. , Rao L. Vijaya Mohan
Published: January 2004

Tissue factor (TF), the cellular receptor for factor VIIa (FVIIa), besides initiating blood coagulation, is believed to play an important role in tissue repair, inflammation, angiogenesis, and tumor metastasis. Like TF, the chemokine interleukin-8 (IL-8) is shown to play a critical role in these processes. To elucidate the potential mechanisms by which TF contributes to tumor invasion and metastasis, we investigated the effect of FVIIa on IL-8 expression and cell migration in a breast carcinoma cell line, MDA-MB-231, a cell line that constitutively expresses abundant TF. Expression of IL-8 mRNA in MDA-MB-231 cells was markedly up-regulated by plasma concentrations of FVII or an equivalent concentration of FVIIa (10 nM). Neither thrombin nor other proteases involved in hemostasis were effective in stimulating IL-8 in these cells. Increased transcriptional activation of the IL-8 gene is responsible for increased expression of IL-8 in FVIIa-treated cells. PAR-2–specific antibodies fully attenuated TF-FVIIa–induced IL-8 expression. Additional in vitro experiments showed that TF-FVIIa promoted tumor cell migration and invasion, active site–inactivated FVIIa, and specific antibodies against TF, PAR-2, and IL-8 inhibited TF-FVIIa–induced cell migration. In summary, the studies described herein provide insight into how TF may contribute to tumor invasion. (Blood. 2004;103:3029-3037)
Serum KIT and KIT ligand levels in patients with gastrointestinal stromal tumors treated with imatinib
Authors: Bono Petri , Krause Andreas , von Mehren Margaret , Heinrich Michael C. , Blanke Charles D. , Dimitrijevic Sasa , Demetri George D. , Joensuu Heikki
Published: January 2004

Imatinib mesylate is a selective inhibitor of a few tyrosine kinases including KIT, and it is the first effective treatment for gastrointestinal stromal tumors (GISTs). We monitored the serum levels of KIT, KIT ligand (stem cell factor, SCF), and the vascular endothelial growth factor (VEGF) in patients with advanced GISTs treated with imatinib in a prospective randomized trial. Patients with GISTs (n = 66) had elevated pretreatment serum KIT and VEGF levels as compared with controls (median, 292 AU/mL [409 ng/mL] vs 238 AU/mL [333 ng/mL], P = .037; and median, 303 pg/mL vs 190 pg/mL, P = .013, respectively), but lower levels of SCF (median, 645 pg/mL vs 950 pg/mL; P ≤ .0001). After 1 and 6 months of imatinib treatment the average serum KIT levels decreased 31% and 52% from pretreatment levels, whereas SCF levels increased 11% and 33%, respectively. Serum VEGF levels decreased during treatment in responding patients. The median serum SCF/KIT ratio increased with treatment duration, and was 7.7-fold higher after 12 months of treatment than at baseline (range, 3.1-259-fold). A high serum SCF/KIT ratio may increase SCF-induced cell signaling with prolonged imatinib treatment, at the time when imatinib treatment is withdrawn, and in patients whose GIST has wild-type receptors. (Blood. 2004;103:2929-2935)
Circulating endothelial cells as a marker of endothelial damage in allogeneic hematopoietic stem cell transplantation
Authors: Woywodt Alexander , Scheer Johanna , Hambach Lothar , Buchholz Stefanie , Ganser Arnold , Haller Hermann , Hertenstein Bernd , Haubitz Marion
Published: January 2004

Damage to endothelial cells is the common feature of vascular disorders associated with hematopoietic stem cell transplantation (HSCT). Elevated numbers of circulating endothelial cells reflect the extent of endothelial damage in a variety of disorders but their use in HSCT has not been investigated so far. We studied 39 patients undergoing allogeneic HSCT with different conditioning regimens and 22 healthy controls. Circulating endothelial cells were enumerated with immunomagnetic isolation during the course of HSCT. After conditioning, cell numbers were significantly elevated (median 44 cells/mL) compared with baseline (median 16 cells/mL) and controls (median 8 cells/mL). Patients who received radiation had an earlier peak when compared with patients who received chemotherapy. Patients who received reduced-intensity conditioning had significantly lower cell numbers (median 24 cells/mL) than those who received standard conditioning. These observations provide a novel marker to investigate microvascular endothelial damage and the effects of different conditioning regimens in patients undergoing HSCT. (Blood. 2004;103:3603-3605)
Inhibition of fibroblast growth factor receptor 3 induces differentiation and apoptosis in t(4;14) myeloma
Authors: Trudel Suzanne , Ely Scott , Farooqi Yildiz , Affer Maurizio , Robbiani Davide F. , Chesi Marta , Bergsagel P. Leif
Published: January 2004

We have previously shown that dysregulation of fibroblast growth factor receptor 3 (FGFR3) by the t(4;14) translocation is a primary event in multiple myeloma (MM) and that activating mutations of FGFR3 are acquired in some cases. We describe here inhibition of wild-type (WT) and constitutively activated mutant FGFR3 autophosphorylation by the small molecule inhibitor, PD173074. Inhibition of FGFR3 in human myeloma cell lines was associated with decreased viability and tumor cell growth arrest. Further, morphologic, phenotypic, and functional changes typical of plasma cell (PC) differentiation, including increase in light-chain secretion and expression of CD31, were observed and this was followed by apoptosis. Finally, using a mouse model of FGFR3 myeloma, we demonstrate a delay in tumor progression and prolonged survival of mice treated with PD173074. These results indicate that inhibition of FGFR3, even in advanced disease associated with multiple genetic changes, may allow the cell to complete its developmental program and render it sensitive to apoptotic signals. In addition, this represents the validation of a therapeutic target in MM that may benefit patients who have a very poor prognosis with currently available treatments. (Blood. 2004;103:3521-3528)
Evaluation of alpha hemoglobin stabilizing protein (AHSP) as a genetic modifier in patients with β thalassemia
Authors: Viprakasit Vip , Tanphaichitr Voravarn S. , Chinchang Worrawut , Sangkla Pakarat , Weiss Mitchell J. , Higgs Douglas R.
Published: January 2004

Although β thalassemia is considered to be a classic monogenic disease, it is clear that there is considerable clinical variability between patients who inherit identical β globin gene mutations, suggesting that there may be a variety of genetic determinants influencing different clinical phenotypes. It has been suggested that variations in the structure or amounts of a highly expressed red cell protein (alpha hemoglobin stabilizing protein [AHSP]), which can stabilize free α globin chains in vitro, could influence disease severity in patients with β thalassemia. To address this hypothesis, we studied 120 patients with Hb E-β thalassemia with mild, moderate, or severe clinical phenotypes. Using gene mapping, direct genomic sequencing, and extended haplotype analysis, we found no mutation or specific association between haplotypes of AHSP and disease severity in these patients, suggesting that AHSP is not a disease modifier in Hb E-β thalassemia. It remains to be seen if any association between AHSP and clinical severity is present in other population groups with a high frequency of β thalassemia. (Blood. 2004;103:3296-3299)
Very low levels of donor CD18+ neutrophils following allogeneic hematopoietic stem cell transplantation reverse the disease phenotype in canine leukocyte adhesion deficiency
Authors: Bauer Thomas R. , Creevy Kate E. , Gu Yu-chen , Tuschong Laura M. , Donahue Robert E. , Metzger Mark E. , Embree Lisa J. , Burkholder Tanya , Bacher John D. , Romines Chris , Thomas Marvin L. , Colenda Lyn , Hickstein Dennis D.
Published: January 2004

Children with the severe phenotype of the genetic immunodeficiency disease leukocyte adhesion deficiency or LAD experience life-threatening bacterial infections because of molecular defects in the leukocyte integrin CD18 molecule and the resultant failure to express the CD11/CD18 adhesion molecules on the leukocyte surface. Hematopoietic stem cell transplantation remains the only definitive therapy for LAD; however, the degree of donor chimerism and particularly the number of CD18+ donor-derived neutrophils required to reverse the disease phenotype are not known. We performed nonmyeloablative hematopoietic stem cell transplantations from healthy matched littermates in 9 dogs with the canine form of LAD known as CLAD and demonstrate that in the 3 dogs with the lowest level of donor chimerism, less than 500 CD18+ donor-derived neutrophils/μL in the peripheral blood of the CLAD recipients resulted in reversal of the CLAD disease phenotype. These results demonstrate the value of a disease-specific, large-animal model for identifying the lowest therapeutic level required for successful cellular and gene therapy. (Blood. 2004;103:3582-3589)
The Second Murine Autoantibody Workshop
Authors: YU LIPING , EISENBARTH GEORGE , BONIFACIO EZIO , THOMAS JAMES , ATKINSON MARK , WASSERFALL CLIVE
Published: January 2004

Abstract: In October 2000, the First Murine Autoantibody Workshop was held as part of an International Workshop on Lessons from Animal Models for Human Type 1 Diabetes. This first workshop identified insulin, but not glutamic acid decarboxylase (GAD) or IA‐2, as specific autoantigens of humoral immunity in nonobese diabetic (NOD) mice. The goals of the Second Murine Autoantibody Workshop, part of the Sixth Annual Meeting of the IDS, were to increase the number of participating investigators, attempt standardization of insulin autoantibody (IAA) results across laboratories, identify serologic evidence of humoral immunity to other beta cell antigens, and allow for validation of ELISA assays for autoantibody detection in NOD mice. Sixty‐three coded samples (26 pooled NOD sera, 23 pooled C57BL/6 sera, and 14 diluted samples of an anti‐insulin monoclonal antibody) were distributed to 12 participating laboratories. This second workshop demonstrated that, for nearly all laboratories, IAA measured by radioimmunoassay (RIA) provided a sensitive and specific assay capable of distinguishing diabetes‐prone from nondiabetes‐prone mice. Analyses involving the serially diluted anti‐insulin monoclonal antibody offered hope that a standard reference unit for reactivity could be established. Surprisingly, two ELISA assays for IAA detection proved remarkably sensitive (i.e., 65% and 92%). However, subsequent absorption studies performed after the workshop (presented at the IDS meeting) brought into question whether ELISA assays for IAA do, in reality, detect anti‐insulin immunities and whether assays for GAD and IA‐2 autoantibodies distinguish diabetes‐prone from nondiabetes‐prone mice. In sum, this workshop continued to support the notion that IAA, as determined by RIA, could provide a sensitive and specific marker of anti‐beta cell immunity in NOD mice.
Can Enteroviruses Cause Type 1 Diabetes?
Authors: TAURIAINEN SISKO , SALMINEN KIMMO , HYÖTY HEIKKI
Published: January 2004

Abstract: Enterovirus infections have long been considered as one possible environmental trigger of type 1 diabetes. These viruses have been detected from diabetic patients more often than from control subjects and they can infect beta cells in cell culture and induce diabetes in animal models. Furthermore, a same kind of seasonality has been observed in both the onset of clinical diabetes and subclinical beta cell autoimmunity (appearance of autoantibodies) as in enterovirus infections. Recently, considerable new evidence has cumulated from prospective studies indicating the risk effect of enterovirus infections long before clinical diabetes was diagnosed. In addition, several studies have reported enterovirus genome sequences in diabetic patients more often than in control subjects. Currently, the evidence for the role of enteroviruses is stronger than for most other environmental agents, but still the final proof is lacking. The ongoing studies aim to prove the risk effect in different populations and to identify the underlying mechanisms. This research field is becoming more and more important because it could open up possibilities to prevent type 1 diabetes by an enterovirus vaccine.
Establishing Evidence for Enterovirus Infection in Chronic Disease
Authors: OBERSTE M. STEVEN , PALLANSCH MARK A.
Published: January 2004

Abstract: Viruses have long been considered among potential environmental triggers of type 1 diabetes mellitus. Epidemiologic and seroprevalence studies have associated enterovirus infection with development of prediabetic autoimmunity and with the onset of clinical diabetes. Enterovirus infection has also been temporally correlated with disease onset by virus isolation or by detection of viral genome by reverse transcription‐polymerase chain reaction (RT‐PCR). For the large‐scale prospective studies that are required to firmly establish a causal relationship between enterovirus infection and development of prediabetic autoimmunity or progression from autoimmunity to clinical diabetes, sensitive RT‐PCR methods must be used to detect virus prior to the onset of diabetic symptoms. We have developed an RT‐seminested PCR protocol to detect enteroviruses in clinical specimens. This method is approximately 10,000‐fold more sensitive than conventional, single‐amplification PCR. Further, we have developed molecular methods to rapidly and reliably identify enterovirus serotype, bypassing the cumbersome and often problematic neutralization test. The molecular serotyping approach will be valuable in examining the relationships between particular virus serotypes or genotypes and specific diseases.
Beta Cell Death and Protection
Authors: MANDRUP‐POULSEN THOMAS
Published: January 2004

Abstract: Type 1 diabetes is an immune‐mediated disease critically dependent upon the interaction between antigen‐presenting cells and T cells. Clearly, both CD4+ and CD8+ T cells are required, but activated CD4+ T cells are both necessary and sufficient in causing disease. The mechanism of the Th1/Th2 immunoregulatory imbalance is unclear and needs to be further investigated. CD8+ T cells are not commonly sufficient in causing disease, but CD8 T cells are necessary in initiation (<14 weeks in the NOD mouse), but not in the later (>14 weeks) effector phase of the disease. It is still unclear whether the CD8+ T cell exerts its function as a classical effector cell or mainly as an immunomodulatory cell acting in synergy with the CD4+ T cell. The relative role of T cell effector mechanisms such as Fas/FasL, perforin/granzyme, and the TRAIL systems is unclear. Proinflammatory cytokines, reactive oxygen species, and other immune mediators seem to be involved in beta cell destruction, but much is to be learned about signaling, molecular mechanisms, and in vivo importance.
Oxidative Stress in Type 1 Diabetes
Authors: HASKINS KATHRYN , BRADLEY BRENDA , POWERS KATHERINE , FADOK VALERIE , FLORES SONIA , LING XIAOFENG , PUGAZHENTHI SUBBIAH , REUSCH JANE , KENCH JENNIFER
Published: January 2004

Abstract: We have been investigating the effects of preventing oxidative stress on pathogenesis and complications of type 1 diabetes in the NOD mouse model. Our studies have shown that damage caused by oxidative stress is higher in islets and vascular tissue of NOD mice than in nonautoimmune controls or a diabetes‐resistant NOD mouse. In addition, phagocytic function and cytokine production by macrophages are aberrant in the NOD. We have demonstrated that treatment of prediabetic NOD mice for 2 weeks with a metalloporphyrin superoxide dismutase (SOD) mimetic results in marked reduction of oxidative stress in islets and vascular tissue and a reversal of macrophage defects.
Use of Microarray Analysis to Unveil Transcription Factor and Gene Networks Contributing to β Cell Dysfunction and Apoptosis
Authors: EIZIRIK DECIO L. , KUTLU BURAK , RASSCHAERT JOANNE , DARVILLE MARTINE , CARDOZO ALESSANDRA K.
Published: January 2004

Abstract: The β cell fate following immune‐mediated damage depends on an intricate pattern of dozens of genes up‐ or downregulated in parallel and/or sequentially. We are utilizing microarray analysis to clarify the pattern of gene expression in primary rat β cells exposed to the proapoptotic cytokines, IL‐1β and/or IFN‐γ. The picture emerging from these experiments is that β cells are not passive bystanders of their own destruction. On the contrary, β cells respond to damage by activating diverse networks of transcription factors and genes that may either lead to apoptosis or preserve viability. Of note, cytokine‐exposed β cells produce and release chemokines that may contribute to the homing and activation of T cells and macrophages during insulitis. Several of the effects of cytokines depend on the activation of the transcription factor, NF‐κB. NF‐κB blocking prevents cytokine‐induced β cell death, and characterization of NF‐κB‐dependent genes by microarray analysis indicated that this transcription factor controls diverse networks of transcription factors and effector genes that are relevant for maintenance of β cell differentiated status, cytosolic and ER calcium homeostasis, attraction of mononuclear cells, and apoptosis. Identification of this and additional “transcription factor networks” is being pursued by cluster analysis of gene expression in insulin‐producing cells exposed to cytokines for different time periods. Identification of complex gene patterns poses a formidable challenge, but is now technically feasible. These accumulating evidences may finally unveil the molecular mechanisms regulating the β cell “decision” to undergo or not apoptosis in early T1DM.
The T Cell Response to Glutamic Acid Decarboxylase 65 in T Cell Receptor Transgenic NOD Mice
Authors: McDEVITT HUGH
Published: January 2004

Abstract: Using BW 5147 T cell hybridomas isolated by fusion with spleen and lymph node cells from NOD female mice, two T cell receptor transgenic NOD mouse lines were produced. Both TCR transgenics respond to their cognate peptide/MHC (GAD65 206‐220 and 286‐300) and produce IL‐2, IFN‐γ, and small amounts of IL‐10. Unexpectedly, the transgenic mice do not develop diabetes and have no insulitis. Analysis with a GAD65 286‐300/I‐Ag7 tetramer reveals that transgenic T cells are negatively selected in the thymus and further negatively selected in the periphery. When crossed to the Cα−/− NOD line, CD4 T cells were reduced by 90% in the thymus and periphery. Further, the tetramer positive GAD65 286‐300 specific T cells were capable of delaying the onset of diabetes in a standard transfer system. Thus, GAD65 specific TCR transgenic T cells (1) must express a second a chain to survive negative selection, (2) produce IL‐2 and IFN‐γ, and (3) have a mildly protective effect on transfer of diabetes with diabetogenic spleen cells.
Detection of CD4+ Autoreactive T Cells in T1D Using HLA Class II Tetramers
Authors: REIJONEN HELENA , KWOK WILLIAM W. , NEPOM GERALD T.
Published: January 2004

Abstract: The use of soluble class II MHC tetramers has enabled the identification of autoantigen‐specific T cells in the peripheral blood of type 1 diabetes patients. Our approach takes advantage of the appearance of highly activated T cells expressing a CD25+/CD4high+ phenotype induced by immobilized class II MHC monomer containing the GAD65 peptide. Almost all T cells that stain with the specific tetramer reside in this population and, since this activation profile is not present in normal subjects, it may provide a useful tool for analysis of the T cell response in autoimmune diabetes. The utilization of tetramer techniques in the detection of autoreactive T cells is a powerful tool to gain insight into mechanisms of the molecular basis of autoimmunity. The phenotyping of T cells should provide useful markers for progression of immune‐mediated β cell reactivity and can be utilized in clinical trials to evaluate the efficacy of the immunomodulatory therapies targeting intervention/prevention of autoimmune diseases.
Kinetic Evolution of a Diabetogenic CD8+ T Cell Response
Authors: SANTAMARIA PERE
Published: January 2004

Abstract: Nonobese diabetic (NOD) mice develop overt diabetes following prolonged periods of pancreatic islet inflammation involving both CD4+ and CD8+ T cells. The initiation and progression of autoimmune diabetes require the recruitment of beta cell‐reactive CD8+ T cells to the pancreatic lymph nodes, their activation by antigen, and their subsequent migration into pancreatic islets. We and others have shown that a significant fraction of NOD islet‐associated CD8+ T cells express highly homologous TCRα chains (Vα17 and Jα42 joined by the same N‐region sequence) and that they recognize the peptide NRP‐A7 in the context of the MHC class I molecule H‐2Kd. We have also shown that this T cell subpopulation undergoes a process of “avidity maturation” that is associated with progression of benign insulitis to overt diabetes. This paper will summarize our current understanding of the mechanisms that drive the recruitment and activation of this CD8+ T cell subpopulation.
Multiple and High‐Titer Single Autoantibodies in Schoolchildren Reflecting the Genetic Predisposition for Type 1 Diabetes
Authors: SCHLOSSER MICHAEL , WASSMUTH RALF , STREBELOW MARTINA , RJASANOWSKI ILONA , ZIEGLER MANFRED
Published: January 2004

Abstract: The study aimed to compare the HLA specificities of AAb‐positive healthy schoolchildren with those of patients with type 1 diabetes (T1D). HLA‐DRB1 and DQB1 alleles were determined in 178 AAb‐positive and 339 AAb‐negative schoolchildren aged 6‐17 years without first‐degree relatives with T1D and in 274 patients with T1D. AAbs against glutamic acid decarboxylase (GADA), protein tyrosine phosphatase (IA‐2A), and insulin (IAA) were determined by 125I‐antigen binding, and islet cell cytoplasmic antibodies (ICAs) immunohistochemically. Here, 82.6% (147/178) of AAb‐positive schoolchildren had single AAbs and 17.4% (31/178) had multiple AAbs. In both groups, GADA occurred with highest and IAA with lowest frequency. In children with single AAbs at levels between the 99th and 99.9th percentile, frequencies of the diabetes‐associated DRB1 (*03, *04) and DQB1 (*02, *0302) alleles and the protective DRB1 (*15) and DQB1 (*0602) alleles did not differ from those of controls. In patients, the positive associations were confirmed for DRB1*04 (OR = 5.39) and DQB1*0302 (OR = 9.05), whereas DRB1*15 (OR = 0.05) and DQB1*0602 (OR = 0.06) were negative‐associated (p < 0.001). The same association was found in schoolchildren with multiple AAbs for DRB1*04 (OR = 3.84), DQB1*0302 (OR = 4.95), and DRB1*15 (OR = 0.1; p < 0.001‐0.014), and with high‐titer single AAbs (≥99.9th percentile), but none of them had DQB1*0602. The highest risk genotype DQB1*02/*0302 occurred in 36.5% of patients (OR = 21.07) and in 19.3% of children with multiple AAbs (OR = 8.8; p<0.001). It is concluded that probands with multiple and high‐titer single AAbs in the general population have the same genetic predisposition for T1D as patients and are therefore at highest risk for the disease.
Insulin Autoimmunity
Authors: EISENBARTH GEORGE S.
Published: January 2004

Abstract: We can now predict the development of type 1A diabetes in humans and prevent the disorder in animal models, but we cannot at present safely prevent type 1A diabetes in humans, although a series of clinical trials are under way and planned. A major lack in our current trial design is the inability to measure T lymphocytes directly responsible for beta cell destruction. Given the immunogenetics of type 1A diabetes and increasing knowledge of pathogenesis in the NOD mouse, we believe the disorder results from immune reactivity to a limited set of islet peptides, with reactivity to insulin a major determinant of disease. Insulin autoantibodies precede the development of diabetes in both humans and the NOD mouse. T lymphocytes isolated from the islets of the NOD mouse that recognize insulin peptide B:9‐23 can transfer diabetes. Insulin expression within the thymus is correlated with genetic susceptibility, and insulin peptides can be used to induce diabetes and as an immunologic vaccine to prevent the disorder. Nevertheless, at present, routine measurement of anti‐insulin T lymphocytes is not standardized. Better assays to monitor such autoreactivity are likely to be essential for the development and evaluation of preventive therapies.
MHC‐Linked Susceptibility to Type 1 Diabetes
Authors: WUCHERPFENNIG KAI W.
Published: January 2004

Abstract: The MHC represents the most important susceptibility locus for type 1 diabetes, and the MHC class II alleles that encode DQ8 and DQ2 in humans and I‐Ag7 in NOD mice represent critical elements. Even though these genetic facts have been known for a number of years, the biochemical and structural features of these MHC class II molecules have only been elucidated recently. We expressed DQ8 and I‐Ag7 as soluble proteins and observed significant structural and functional similarities between these human and murine MHC molecules. It had been postulated that I‐Ag7 and DQ8 are poor peptide binders, based on the observation that the subunits dissociate in the presence of SDS, a denaturing detergent. We observed that both DQ8 and I‐Ag7 form stable, long‐lived complexes with a number of different peptides, indicating that they are not unstable in the absence of detergent. DQ8 and I‐Ag7 were found to bind similar sets of peptides, which included peptides that had been identified as immunodominant T cell epitopes of insulin and GAD 65 in NOD mice. The insulin B (9‐23) peptide formed long‐lived complexes with DQ8 and was thus chosen for crystallization of the complex. No defect in the peptide binding groove was evident in the crystal structure and the insulin peptide was deeply anchored in the binding site. The structure demonstrated significant similarities between DQ8 and I‐Ag7, indicating that similar antigen presentation events are relevant in the NOD mouse model and the human disease.
Using Regulatory APCs to Induce/Maintain Tolerance
Authors: JUEDES AMY E. , VON HERRATH MATTHIAS G.
Published: January 2004

Abstract: It is well established that antigen‐presenting cells (APCs) such as dendritic cells (DCs) possess potent immune‐stimulatory function. They are considered to be the key driving element for most immune, autoimmune, and host‐defense responses. However, recent evidence suggests that as much as APCs can turn things on, they also have the capability to turn things off. To summarize the evidence for such regulatory function of APCs is the purpose of this review article. We are just beginning to understand whether regulatory APC function can be mapped to a separate lineage of APCs or whether it is more commonly acquired under certain maturation conditions. Furthermore, it becomes apparent that it is important to consider differences between human and mouse DCs as well as splenic‐, lymph node‐, or blood‐derived APCs. We believe that, in the upcoming years, better understanding of positive and negative roles of APCs in immune regulation will benefit both sides of APC therapy: their use in enhancing immunity, for example, in vaccine design and cancer, as well as their application for the treatment of autoimmunity.
Cell‐Based Therapies for Diabetes: Progress towards a Transplantable Human β Cell Line
Authors: ITKIN‐ANSARI PAMELA , GERON IFAT , HAO ERGENG , DEMETERCO CARLA , TYRBERG BJORN , LEVINE FRED
Published: January 2004

Abstract: Achieving normoglycemia is the goal of diabetes therapy. Potentially, there are many ways to achieve this goal, including transplantation of cells exhibiting glucose‐responsive insulin secretion. However, to be applicable to the large number of people who might benefit from β cell replacement, an unlimited supply of β cells must be found. To address this problem, we have been developing cell lines from the human endocrine pancreas. In one case, a cell line, βlox5, has been developed from human islets that can be induced under some circumstances to differentiate into functional β cells exhibiting appropriate glucose‐responsive insulin secretion. Inducing differentiation is complex, requiring the activation of multiple signaling pathways, including those downstream of those involved in cell‐cell contact and the glucagon‐like peptide‐1 receptor. In addition, transfer of the PDX‐1 gene is also necessary to render the cells competent for differentiation. However, it is clear that many other genes are involved in maintaining the commitment of βlox5 cells towards the β cell lineage. Understanding the complement of genes required to establish and maintain a β cell lineage commitment would be enormously helpful in efforts to develop a cell line that can be used for β cell replacement therapies. Here, we provide further information on the characteristics of cell lines that we have developed from the human pancreas that are relevant to the development of a β cell replacement therapy for diabetes.
Islet Cell Autoimmunity and Transplantation Tolerance: Two Distinct Mechanisms?
Authors: PEARSON TODD , MARKEES THOMAS G. , SERREZE DAVID V. , PIERCE MELISSA A. , WICKER LINDA S. , PETERSON LAURENCE B. , SHULTZ LEONARD D. , MORDES JOHN P. , ROSSINI ALDO A. , GREINER DALE L.
Published: January 2004

Abstract: Recent advances in islet transplantation have enabled physicians to cure type 1 autoimmune diabetes, but at the cost of lifelong immunosuppression with its attendant side effects and long‐term health risks. To eliminate the need for immunosuppression, researchers have developed methods for inducing tolerance to transplanted allografts. Tolerance‐based transplantation using costimulation blockade has proven remarkably successful in many animal model systems. The most widely used animal model system for studying islet transplantation in type 1‐like autoimmune diabetes is the NOD mouse. Unfortunately, this strain has proven resistant to costimulation blockade‐based transplantation tolerance protocols that are successful in chemically diabetic mice given islet grafts. It has been assumed that resistance to transplantation tolerance in the NOD mouse is (1) related to autoimmunity directed against its pancreatic β cells, (2) a consequence of that autoimmunity, and (3) under the control of the same genes that control autoimmunity. In this review, we provide arguments to challenge these assumptions. We describe a new animal model and a new conceptual framework based on data indicating that the mechanisms responsible for resistance to transplantation tolerance and β cell autoimmunity are not identical. We believe that the recent discoveries we describe will have important implications for the development of tolerance‐based transplantation therapies and their translation from the laboratory to the clinic.
Characterization of Early Developments in the Splenic Leukocyte Transcriptome of NOD Mice
Authors: GERLING I. C. , ALI C. , LENCHIK N.
Published: January 2004

Abstract: Transcriptome analysis is a powerful tool to characterize changes in leukocyte gene expression patterns and reveal very early molecular abnormalities in tissue. Herein, we report on characterization of the very earliest abnormalities in the transcriptome of leukocytes from young “prepathologic” NOD and NON female mice.
Autoimmune Diabetes in the NOD Mouse: An Essential Role of Fas‐FasL Signaling in β Cell Apoptosis
Authors: SILVA DIEGO G. , SOCHA LUIS , CHARLTON BRETT , COWDEN WILLIAM , PETROVSKY NIKOLAI
Published: January 2004

Abstract: Despite evidence that both Fas and FasL can be expressed in pancreatic islets, there has been considerable controversy regarding the potential role of Fas signaling in autoimmune β cell death. Using the HIPFasL model, we have been able to demonstrate that, in the presence of an inflammatory infiltrate, FasL‐expressing β cells are exquisitely sensitive to Fas‐mediated apoptosis and that this can be blocked by preventing FasL‐Fas interaction. This points to a highly important role of Fas‐FasL interaction in autoimmune β cell death.
Fas and Fas Ligand Immunoexpression in Pancreatic Islets of NOD Mice during Spontaneous and Cyclophosphamide‐Accelerated Diabetes
Authors: REDDY SHIVA , ROSS JACQUELINE M.
Published: January 2004

Abstract: During insulin‐dependent diabetes mellitus, beta cell destruction may involve activation of the Fas‐Fas ligand (Fas‐FasL) system. Here, we employed dual‐label immunohistochemistry to examine the intra‐islet expression, distribution, and cellular sources of Fas and FasL in the NOD mouse. Pancreatic tissues were studied during spontaneous diabetes (days 21, 40, and 90) and following acceleration of diabetes with cyclophosphamide (days 0, 4, 7, 11, and 14 after cyclophosphamide administration). Our results show that FasL was expressed constitutively in most beta cells of NOD mice and in nondiabetes‐prone mice, but not in glucagon or somatostatin cells or in islet inflammatory cells. It paralleled the loss of insulin immunolabeling with advancing disease. Immunolabeling for Fas was first observed in extra‐islet macrophages and those close to the islet in NOD and nondiabetes‐prone mice. During spontaneous and cyclophosphamide diabetes, it was observed in a higher proportion of islet infiltrating macrophages than in CD4 and CD8 cells. In the cyclophosphamide group, Fas expression in intra‐islet CD4 and CD8 cells showed an increase close to the onset of diabetes. At days 11 and 14, several intra‐islet macrophages with immunolabeling for Fas also coexpressed interleukin‐1β and inducible nitric oxide synthase. Fas was not detected in beta cells and other endocrine cells during spontaneous and cyclophosphamide diabetes. We show constitutive expression of FasL in beta cells in the NOD mouse and predominant expression of Fas in intra‐islet macrophages and to a lesser extent in T cells prior to diabetes onset. The role of Fas‐FasL in beta cell destruction in the NOD mouse requires further clarification.
Type 1 Diabetes in Swedish Bank Voles (Clethrionomys glareolus): Signs of Disease in Both Colonized and Wild Cyclic Populations at Peak Density
Authors: NIKLASSON BO , HÖRNFELDT BIRGER , NYHOLM ERIK , NIEDRIG MATTHIAS , DONOSO‐MANTKE OLIVER , GELDERBLOM HANS R. , LERNMARK ÅKE
Published: January 2004

Abstract: Colonized bank voles (Clethrionomys glareolus) originating from Sweden developed type 1 diabetes. Animals became polydipsic, glucosuric, and hyperglycemic and gradually developed a lethal ketoacidosis. Pancreas in animals with end‐stage disease showed total destruction of islet cells. Interestingly, also a high proportion of wild bank voles in cyclic populations that were trapped at (or close to) the cyclic population density peak frequently showed high blood glucose levels and pathological glucose tolerance test. Extensive islet destruction was not seen in wild bank voles at the time of capture, but did develop in some of the animals over a time period of two months. Diabetes in both colonized and wild bank voles was associated with Ljungan virus (LV). LV could be isolated from the pancreas of diabetic bank voles and antigen detected at the site of tissue damage by immunohistochemistry. In addition, picornavirus‐like particles were visualized in the islets of diabetic voles using thin‐section transmission electron microscopy.
Defective Activation‐Induced Cell Death in NOD T Lymphocytes
Authors: DECALLONNE B. , MATHIEU C.
Published: January 2004

Abstract: The aim of this study was first to analyze strain differences in apoptosis resistance of NOD peripheral T lymphocytes in in vitro models of death by neglect and activation‐induced cell death (AICD). Especially AICD is known to play a key role in peripheral tolerance, keeping autoimmune effector cells under control. Second, we studied the effects of in vivo treatment of NOD mice with 1α,25‐dihydroxyvitamin D3 [1α,25(OH)2D3], an immunomodulator known to prevent diabetes and insulitis in NOD mice, on in vitro T lymphocyte apoptosis resistance.
The Role of Endoplasmic Reticulum Stress in Nonimmune Diabetes: NOD.k iHEL, a Novel Model of β Cell Death
Authors: SOCHA L. , SILVA D. , LESAGE S. , GOODNOW C. , PETROVSKY N.
Published: January 2004

Abstract: The final common pathway in diabetes development is β cell apoptosis. We herein describe a novel diabetes model based on transgenic NOD.k iHEL mice, wherein male mice develop diabetes due to nonimmune‐mediated β cell death. Histology and electron microscopy confirm endoplasmic reticulum (ER) abnormalities that are consistent with endoplasmic stress caused by the HEL transgene. The NOD.k iHEL model may be particularly useful for studying mechanisms of β cell death secondary to ER stress and also for testing potential therapies designed to protect β cells from stress‐induced apoptosis. The observation that only male NOD.k iHEL mice develop diabetes and exhibit ER abnormalities is intriguing and suggests these mice may be useful in deciphering the link between hyperandrogenism, insulin resistance, and diabetes.
Defective Maturation of Myeloid Dendritic Cell (DC) in NOD Mice Is Controlled by IDD10/17/18
Authors: PENG RUIHUA , BATHJAT K. , LI Y. , CLARE‐SALZLER M. J.
Published: January 2004

Abstract: We previously demonstrated that adoptive transfer of NOD pancreatic lymph node (PLN) DC protected recipients from diabetes. Our recent studies showed that the tolerogenic DC population presented islet antigens and were mature myeloid DC that did not produce IL‐12, suggestive of exhausted or fully mature DC. Extensive characterization of the DC population in vivo in NOD and control mice demonstrated a specific deficiency of PLN tolerogenic DC in older mice. These findings suggest autoimmunity might arise in NOD mice secondary to deficient maturation of myeloid DC to a tolerogenic state. To address this issue, we characterized maturation and function at development of bone marrow‐derived myeloid DC from NOD and several control strains. We found that NOD DC were highly resistant to several maturation stimuli and maintained an immature phenotype (average % immature DC: 75% in NOD versus 15% in B6, p < 0.01). A survey of congenic NOD mice with various NOD diabetes susceptibility loci demonstrated that the IDD10/17/18 region on chromosome 3 controlled approximately 50% of the NOD DC maturation defect. The defect also affected NOD DC that underwent phenotypic maturation. These cells appeared to arrest in a “maturing” phase as they produced 5‐ to 7‐fold more IL‐12 than control strains and significantly less IL‐10. The cytokine defect was completely corrected in NOD IDD10/17/18 mice. In addition, the IDD10/17/18 locus limited DC accumulation in islets and significantly increased tolerogenic DC in the PLN. Together, the above findings suggest that polygenic regulation of DC maturation defects in NOD mice promotes islet inflammation while limiting the generation of tolerogenic DC.
Absence of the T‐bet Gene Coding for the Th1‐Related Transcription Factor Does Not Affect Diabetes‐Associated Phenotypes in Balb/c Mice
Authors: MELANITOU EVIE , LIU EDWIN , MIAO DONGMEI , YU LIPING , GLIMCHER LAURIE H. , EISENBARTH GEORGE
Published: January 2004

Abstract: The T‐box expressed in T cells gene (T‐bet) is a member of the T‐box family of transcription factors. T‐bet‐deficient mice show normal lymphoid development, but exhibit profound defects in their Th1‐mediated immune responses. As the balance between Th1‐ and Th2‐mediated immune responses plays a role in autoimmune‐prone diseases, we have investigated the diabetes‐related insulin autoantibody (IAA) and cellular immune responses (insulitis), in the absence of Th1 lineage commitment, in T‐bet KO Balb/c mice, after immunization with the B9‐23 insulin peptide. We have therefore investigated whether absence of the T‐bet gene influences diabetes‐related phenotypes in Balb/c T‐bet KO mice.
Immunolocalization of Caspase‐3 in Pancreatic Islets of NOD Mice during Cyclophosphamide‐Accelerated Diabetes
Authors: REDDY SHIVA , BRADLEY JOSHUA , ROSS JACQUELINE M.
Published: January 2004

Abstract: Apoptosis may be a major mechanism of beta cell loss during insulin‐dependent diabetes mellitus. Caspase‐3 is a key enzyme involved in the terminal steps of this death process. Here, the intra‐islet expression of caspase‐3 in the NOD mouse was examined immunohistochemically following acceleration of the disease with cyclophosphamide. Female NOD mice were treated at day 95 with cyclophosphamide, and caspase‐3 expression in pancreatic sections was studied at days 0, 4, 7, 11, and 14 and compared with age‐matched control tissue. In the treated group at day 0, caspase‐3 labeling was seen in several peri‐islet macrophages and only extremely rarely in beta cells. At day 4, only a few beta cells weakly expressed the enzyme. From day 7, caspase‐3 expression began to increase in intra‐islet macrophages and reached a peak at days 11 and 14, when a small number of CD4 and CD8 T cells also showed positive labeling. Beta cell expression of caspase‐3 at days 11 and 14 was rare. At this stage, several intra‐islet immune cells with positive labeling for the enzyme coexpressed either Fas or interleukin‐1β. Only a small proportion of intra‐islet caspase‐3 cells showed apoptotic nuclei judged by terminal deoxynucleotidyl transferase‐mediated dUTP nick end labeling (TUNEL). We conclude that, during cyclophosphamide‐accelerated diabetes, the predominant caspase‐3 immunolabeling in intra‐ and extra‐islet macrophages suggests that apoptosis of macrophages may be an important mechanism for their elimination. The virtual absence of caspase‐3 immunolabeling in most beta cells even during the height of beta cell loss supports the need for developing other markers of early beta cell apoptosis in the NOD mouse.
Congenic Mapping and Candidate Sequencing of Susceptibility Genes for Type 1 Diabetes in the NOD Mouse
Authors: IKEGAMI HIROSHI , FUJISAWA TOMOMI , MAKINO SUSUMU , OGIHARA TOSHIO
Published: January 2004

Abstract: Inheritance of type 1 diabetes is polygenic with a major susceptibility gene located in the major histocompatibility complex (MHC). In addition to MHC‐linked susceptibility, a number of susceptibility genes have been mapped outside the MHC in both humans and animal models. In order to localize and identify susceptibility genes for type 1 diabetes, we have developed a series of congenic strains in which either susceptibility intervals from the NOD mouse, a mouse model of type 1 diabetes, were introgressed onto control background genes or protective intervals from control strains were introgressed onto NOD background genes. NOD. CTS‐H‐2 congenic mice, which possess recombinant MHC with NOD alleles at class II A and E genes, which are candidates for Idd1, revealed that Idd1 consists of multiple components, one in class II (Idd1) and the other adjacent to, but distinct from, Idd1 (Idd16). Phenotypes of NOD. IIS‐Idd3 congenic mice, which share the same alleles at both Il2 and Il21 as the NOD mouse, were indistinguishable from the NOD parental strain, indicating that both Il2 and Il21 are candidates for Idd3. In contrast, NOD. IIS‐Idd10 congenic mice, which share the same alleles at Fcgr1, a previous candidate for Idd10, as the NOD mouse, were protected from type 1 diabetes, suggesting that Fcgr1 may not be responsible for the Idd10 effect. These data suggest that the use of strain colony closely related to a disease model to find the same candidate mutation on different haplotypes and make congenic strains with this recombinant chromosome, termed ancestral haplotype congenic mapping, is an effective strategy for fine mapping and identification of genes responsible for complex traits.
Establishing Insulin 1 and Insulin 2 Knockout Congenic Strains on NOD Genetic Background
Authors: PARONEN JOHANNA , MORIYAMA HIROAKI , ABIRU NORIO , SIKORA KAMILA , MELANITOU EVIE , BABU SUNANDA , BAO FEI , LIU EDWIN , MIAO DONGMEI , EISENBARTH GEORGE S.
Published: January 2004

Abstract: As insulin is a major autoantigen in autoimmune diabetes and because the insulin gene region locus in humans has been linked to diabetes risk, we have bred insulin gene knockouts onto the NOD mouse. Mice differ from humans in terms that they express two nonallelic genes of insulin. Insulin 2 is the murine homologue of the human insulin gene and is located on mouse chromosome 7. Insulin 1 is thought to have evolved by a gene duplication event, lacks the second intron of the insulin 2 gene, and is located on mouse chromosome 19. The differential thymic expression of the insulin gene may be important for central tolerance induction. Here, we present the initial establishment of congenic knockouts and characterization of the congenic intervals corresponding to insulin 1 and insulin 2 knockout genes on mouse chromosome 19 and 7, respectively.
Relationship between β Cell Mass of NOD Donors and Diabetes Development of NOD‐scid Recipients in Adoptive Transfer System
Authors: YAMADA SATORU , SHIMADA AKIRA , KODAMA KEIICHI , MORIMOTO JIRO , SUZUKI RYUJI , OIKAWA YOICHI , IRIE JUNICHIRO , SARUTA TAKAO
Published: January 2004

Abstract: Background—There is controversy about the way the β cell mass is reduced in type 1 diabetes. One view is that a gradual fall in β cell mass begins soon after the onset of insulitis. Another view is that a sudden wave of β cell destruction occurs just before the onset of diabetes. To clarify how the β cell mass is reduced, we performed adoptive transfer experiments and examined the relationship between the pancreatic status of NOD donors and the time taken to transfer diabetes into NOD‐scid recipients. Methods—We killed 18‐week‐old female NOD mice (n= 20), removed their spleen, and transferred splenocytes into 5‐week‐old female NOD‐scid mice (n= 60). The relationship between the pancreatic status of donors and the time taken to transfer diabetes into recipients was assessed. As pancreatic status, we measured insulin content and severity of insulitis. Results—There was no linear correlation between the pancreatic status of donors and the time taken to transfer diabetes into recipients. NOD donors who needed 7 or more weeks to transfer diabetes in NOD‐scid recipients had similar levels of insulin content or severity of insulitis as those of NOD donors who could not transfer diabetes. On the other hand, NOD donors who needed 6 or less weeks to transfer diabetes in recipients had similar levels of insulin content or severity of insulitis as those of diabetic NOD mice. Conclusions—According to our observations, β cell mass seems to be preserved until just before the onset of diabetes and decreased dramatically within a few weeks.
Hen Egg White Lysozyme Vaccination Induces Acute Shock in NOD Mice
Authors: BRANISTEANU DUMITRU D. , OVERBERGH LUTGART , DECALLONNE BRIGITTE , MATHIEU CHANTAL
Published: January 2004

Abstract: To investigate whether vaccination could induce lethal shock and which mechanisms are involved in this phenomenon, we tested a panel of autoantigens or diabetes‐irrelevant peptides or proteins in nonobese diabetic (NOD), Balb/c, and C57Bl/6 mice. Of the antigens tested, only nondiabetogenic hen egg white lysozyme induced a severe form of shock exclusively in NOD mice. The mechanism involved is suggestive of a Th2‐mediated anaphylactic reaction possibly connected to activation of PAF and triggering of DIC.