13 However, the growth cycle can be slowed or arrested depending on intracellular nutrient availability, leading to bacterial persistence within host cells.14,15 This is a key survival feature of these organisms and is a major determinant of disease pathogenesis as discussed more fully in the following sections. C. abortus typically causes reproductive failure and abortion in ruminants and swine and has a world-wide distribution, with the exception of Australia and New Zealand. C. abortus is also a well-recognized and potentially

fatal zoonosis, presenting a major hazard to pregnant women who come in contact with livestock, particularly at lambing.16 Although OEA is a reproductive disease, the principal route of transmission to naïve sheep is thought to be via an oro-nasal route, most likely from heavily infected placentas from ewes that have aborted and contaminate the environment.17,18 A typical example R788 mouse of a placenta with characteristic thickened www.selleckchem.com/products/atezolizumab.html membranes from an ewe that aborted as a result of OEA is shown in Fig. 2. Abortion is thought to be because of inappropriate inflammatory cytokine and chemokine production in the placenta that leads to placentitis.18,19 The success of C. abortus as a reproductive pathogen in a species that is only pregnant for 5 months

and only gives birth once a year is because of its ability to establish a persistent, subclinical infection in non-pregnant sheep.20 Thus, when naïve, non-pregnant sheep are infected, protective immunity does not develop. Ewes then abort in the subsequent pregnancy. Sheep that have aborted do develop strong protective immunity (but not necessarily sterile immunity) and reproduce normally in subsequent pregnancies.20,21 The Adenylyl cyclase epidemiology and pathogenesis of OEA both indicate that a systemic phase of infection occurs after the primary infection of the oro-nasal mucosa. Neither the site of persistence of C. abortus nor the timing or duration of the systemic phase of infection has been identified. Therefore, the paradigms relating to reproductive immunology and to host immune control of intracellular bacteria are useful frameworks for addressing questions regarding

the pathogenesis of OEA. Furthermore, in addressing these paradigms in sheep, we can test their predictions and assess their relevance for a species other than mouse or human. In doing so, we should advance our knowledge of comparative immunology and reproduction. The first description of helper T-cell clones expressing distinctive cytokine profiles was made by Tim Mosmann, Robert Coffman and co-workers22 in 1986 in a paper that has had a profound impact on our understanding of how CD4+ve T cells orchestrate and regulate immune responses. They discovered that mitogen-activated murine CD4+ve T-cell clones were mutually exclusive in their expression of IL-2/IFN-γ (TH1) and what we now know to be IL-4 (TH2), whereas both sets of clones made IL-3.

7a). Induction of IL12p40 expression on rhesus pDC, as observed with mAb C8·6, was confirmed by using another anti-IL-12p40/70 mAb (clone

C11·5), which gave similar percentages of positive cells for pDC as well as mDC upon TLR-7/8 stimulation (Fig. 7b). Finally, analysis of IL-12p40 mRNA in TLR-7/8 (CL097)-stimulated purified pDC, mDC and monocyte populations showed similar high expression levels in pDC relative to mDC and monocytes and no induction in pDC upon TLR-4 stimulation (Fig. 8a), thus confirming the FACS expression data. Both mDC and monocytes up-regulated TNF-α mRNA expression upon TLR-4 (LPS) as well as TLR-7/8 (CL097) stimulation, underscoring the functional capacity of these purified cell populations (Fig. 8b). In agreement with the FACS analysis, TNF-α mRNA expression in pDC was up-regulated only upon TLR-7/8 and not TLR-4 stimulation. While the mDC and pDC preparations were only 60–75% pure the other cells present were find more Dorsomorphin either granulocytes or monocytes

and this could not have affected the IL-12p40 expression data, as monocytes were observed to have only very low IL-12p40 expression and the monocyte fraction itself was >90% pure with <5% mDC and <1% pDC contamination. In this work, we adapted a whole blood stimulation assay to study functional characteristics of peripheral blood DCs and monocytes in macaques and performed a direct comparison with human blood samples. Most responses of the different subsets were very similar between macaques and humans and in agreement with previous studies, in which purified cell populations instead of whole blood stimulation had been used [2, 17, 25-28, 32]. However, we observed that, in contrast to humans, rhesus pDC expressed

IL-12p40 upon stimulation with TLR-7/8 or TLR-9. Preliminary data suggest a similar IL-12p40 expression pattern in cynomolgus macaques (V.S., to be published elsewhere). We also observed that relative to humans, mDC G protein-coupled receptor kinase and monocytes in rhesus macaques responded less well to TLR-7/8 stimulation when expressed as percentage of IL-12p40- and TNF-α-positive cells. Of note is that a similar relatively lower level of IL-12 induction has been reported previously for macaque monocyte-derived DC [23]. The capacity of rhesus pDC to produce IFN-α as well as IL-12p40 may potentially modify their response to viral infections, where pDC are known to play an important role [36]. Previous studies either did not include IL-12 in their analysis [23] or measured IL-12 cytokine production by enzyme-linked immunosorbent assay (ELISA) on either stimulated total PBMC or lineage-negative cell cultures [25-27]. Others used FACS analysis, but studied IL-12 expression only in LPS-stimulated PBMC [17], which would have given no expression in pDC. Hence, our observation was made possible by the use of FACS analysis to detect TLR-induced cytokine expression in all subsets simultaneously.

Various strategies based on modified live or inactivated vaccines have been used to control Aujeszky’s disease. Although a modified live vaccine is known to successfully minimize both the clinical symptoms and viral shedding during the acute phase of PrV infection (13), these strategies still have some disadvantages including the risk of reversion to virulence (13–15) and interference with efficient antigen presentation (15). In contrast, inactivated PrV vaccine is harmless Decitabine cell line but insufficient to induce effective protection against PrV infection. Therefore, the need to

develop a safe vaccine that can induce complete protection against PrV infection remains. We previously demonstrated that attenuated aspartate β-semialdehyde dehydrogenase (Asd)-negative Salmonella enterica serovar Typhimurium devoid of antibiotic resistance gene is an effective delivery system for the mass administration of cytokines without the need for antibiotic selection (16–18). Furthermore, the oral administration of S. enterica serovar Typhimurium expressing cytokines such as chicken IFN-α and IL-18 ameliorated the clinical signs caused by respiratory infection with avian influenza virus (19,20). However, the modulatory effect of the oral co-administration of S. enterica serovar Typhimurium expressing swIFN-α and swIL-18 on the immune responses induced by parenteral administration with inactivated AZD6244 vaccine

was not addressed. Here, we investigated the modulatory effect of the combined administration of swIL-18 and swIFN-α on vaccination with inactivated PrV vaccine using

Salmonella enterica serovar Typhimurium as delivery system. Ultimately, we demonstrate the benefit of the combined administration of swIL-18 and swIFN-α using attenuated S. enterica serovar Typhimurium to provide effective immune responses against the inactivated PrV vaccine. Seronegative crossbreed F1 (Large white-Landrace × Duroc) piglets (3–4 weeks old) were obtained from a local breeding farm and housed in stainless steel cages (2–3 piglets/cage). Piglets were reared with formulated commercial feed and water STK38 provided ad libitum throughout the experimental period. All experimental and animal management procedures were undertaken in accordance with the requirements of the Animal Care and Ethics Committees of Chonbuk National University. The animal facility of the Chonbuk National University is fully accredited by the National Association of Laboratory Animal Care. The wild-type PrV YS strain and thymidine kinase-deleted PrV were generously supplied by the National Veterinary Research and Quarantine Service in the Republic of Korea. The viruses were propagated in the porcine kidney cell line, PK-15, using Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 2.5% fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 U/mL).

In this study we report that the proinflammatory cytokines interleukin (IL)-2, interferon (IFN)-γ and tumour necrosis factor (TNF)-α show a time-dependent increase upon ex-vivo bacterial, viral and fungal antigen stimulations. Furthermore, evidence is provided that this assay is sensitive to mirror stress hormone-mediated immune modulation in humans as shown either after hydrocortisone injection or after acute

stress exposure during free fall in parabolic flight. This in-vitro test appears to be a suitable assay to sensitively mirror stress hormone-dependent inhibition of cellular immune responses in the human. Cell Cycle inhibitor Because of its standardization and relatively simple technical handling, it may also serve as an appropriate research

tool in the field of psychoneuroendocrinology in clinical as in field studies. Humans are continuously subjected to environmental challenges which affect the immune function according to the intensity of psychological and physiological stressors. Due to the complex nature of in-vivo immune responses, the delayed-type hypersensitivity (DTH) skin test has served as a standardized tool to monitor the overall status of the immune system by simultaneously placing six antigens and one diluent (as a negative control) intracutaneously into the forearm. With the DTH skin test it was possible to Saracatinib evaluate, to a certain degree, the extent of immunodeficiency, as seen in individuals infected with the human immunodeficiency virus (HIV) [1].

In addition to being used as a clinical investigative tool in immune deficiency states, the DTH skin test was also used widely to monitor immune function in states of psychological stress and psychiatric illness. Declines in immune function were found in subjects suffering from severe depression [2, 3], in Liothyronine Sodium crews wintering in the Antarctic [4, 5] and individuals experiencing perceived distress [6-9]. In 2002 this in-vivo skin test (multi-test CMI; Mérieux, Lyon, France) was removed from the market, in part because of the risk of antigen-sensitization when applied repeatedly to the same individual. After the DTH skin test was phased out, no such alternative tests were available to evaluate overall immunity. Standardized in-vitro methods such as the lymphocyte transformation test [10] and in-vitro cytokine induction [11] are used for the measurement of antigen-dependent T cell responses, but these tests are complicated in their performance and may not mirror the immune responses to the pathogenic spectrum that the DTH skin test was able to recall. Even though the complex skin reaction of the DTH skin test – which includes, e.g. cell migration – cannot be reproduced fully in a whole-blood in-vitro system, DTH reactions also seem possible to be reflected in blood tests [12, 13].

The coronary arterioles dilated dose-dependently to the endothelium-dependent NO-mediated vasodilator serotonin. This vasodilation was inhibited in the same manner by NOS inhibitor NG-nitro-l-arginine methyl ester and by lumenal OxLDL (0.5 mg protein/mL). The inhibitory effect of OxLDL was reversed after treating the vessels with either l-arginine (3 mM) or arginase inhibitor Selleck MLN8237 difluoromethylornithine (DFMO; 0.4 mM). Consistent with vasomotor alterations, OxLDL inhibited serotonin-induced NO release from coronary arterioles and this inhibition

was reversed by DFMO. Vascular arginase activity was significantly elevated by OxLDL. Immunohistochemical analysis indicated that OxLDL increased arginase I expression in the vascular wall without altering

eNOS expression. Taken together, these results suggest that OxLDL up-regulates arginase I, which contributes to endothelial dysfunction by reducing l-arginine availability to eNOS for NO production and thus vasodilation. “
“Department of Cardiovascular Science, Faculty of Medicine, Dentistry & Health, University of Sheffield, Medical School, Sheffield, UK Atherosclerosis is a chronic inflammatory disease of the medium and large arteries driven in large part by the accumulation of oxidized low-density lipoproteins and other debris at sites rendered susceptible because of the geometry of the arterial tree. As lesions develop, they DNA Damage inhibitor acquire a pathologic microcirculation that perpetuates lesion progression, both by providing a means for further monocyte and T-lymphocyte recruitment into the arterial wall and by the physical and chemical stresses caused by micro-hemorrhage. This review summarizes work performed in our department investigating the roles

of signaling pathways, alone and in combination, that lead to specific programs of gene expression in the atherosclerotic environment. Focusing particularly on cytoprotective responses that might be enhanced therapeutically, the work has encompassed the anti-inflammatory effects of arterial laminar shear stress, mechanisms oxyclozanide of induction of membrane inhibitors that prevent complement-mediated injury, homeostatic macrophage responses to hemorrhage, and the transcriptional mechanisms that control the stability, survival, and quiescence of endothelial monolayers. Lastly, while the field has been dominated by investigation into the mechanisms of DNA transcription, we consider the importance of parallel post-transcriptional regulatory mechanisms for fine-tuning functional gene expression repertoires. “
“Isolation of rodent endothelial cells from lymphatic capillaries with yields that allow extensive functional studies remains challenging due to low cell numbers, variable purity, and limited growth potential.

However, renal biopsies have not revealed adequate information for predicting prognosis. Thus, this retrospective study was conducted of diabetic nephropathy to obtain prognostic information from histopathologic findings. Methods: The subjects were 28 diabetic nephropathy patients confirmed by renal biopsy who were seen between August 2007 and December 2012. Histopathological and clinical findings with renal outcomes were studied. The histopathological scores were determined according to Tervaert et al.: glomerular lesions (score 0–20)

were based on degree of mesangial expansion, GBM thickness, exudative lesion, nodular sclerosis, mesangiolysis, polar vasculosis, global sclerosis, segmental sclerosis, ischemic selleck chemical sclerosis, and hypertrophy; interstitial and tubular lesions (score 0–6) were based on degree of interstitial fibrosis, tubular atrophy, and interstitial inflammation; and vascular lesions (score 0–5) were based on degree of arteriolar hyalinosis and arteriosclerosis. Renal dysfunction was defined as doubling of serum creatinine

concentration, chronic hemodialysis initiation or renal transplantation. Results: Renal survival rates contrasting the low and high score groups in each of the three types of lesions were studied by Kaplan-Meier analysis. The mean survival periods of the low and high score groups for the glomerular CH5424802 (p = 0.24) and vascular (p = 0.22) lesions were not different. However, renal survival rates of 19 and 8 months for the low and high score groups (p = 0.010) respectively, in the interstitial and tubular lesions were significant. Conclusion: Interstitial and tubular lesions were a significant predictor for renal prognosis in diabetic nephropathy. Inasmuch as the histopathology of the glomerulus is known to provide important information of renal disease, our study indicates that significant prognostic information

may be associated with interstitial and tubular changes. MENG XIANJIE1, SHEN SHANMEI2, WAN YIGANG2, LUO XUNYANG2, ZHANG LE2, CHEN HAOLI1, SHI XIMIAO1, HUANG YANRU1, MAO ZHIMIN1 1Department of Graduate School, Nanjing University of Chinese Medicine; 2Nanjing filipin Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School Introduction: Diabetic nephropathy (DN), as a common health problem worldwide, is a dominant cause of end-stage renal disease (ESRD). Therefore, noninvasive detections and dynamic managements in clinic are of major importance of preventing progression from DN to ESRD. The early diagnosis of DN has focused on measurement of urinary albumin (UAlb) excretion rate, but UAlb is actually not overall and sensitive marker for DN patients with inchoate and latent injuries in glomeruli and renal tubules.

Naïve CD4+ T cells were labeled with CFSE and co-cultured with primary Th17 clones, and naïve CD4+ T-cell proliferation was determined

by FACS analysis of CFSE dilutions. As shown in Fig. 1E, we observed that these Th17 clones increased the proliferation of naïve T cells with several cell generations in the presence of OKT3, suggesting that these Th17 clones had effector T-cell function. Furthermore, Th1-C1, a Th1 cell line derived from a melanoma TILs selleckchem which served as an effector T-cell control, also increased the proliferation of naïve T cells. In contrast, a CD4+CD25+ Treg line, which served as a suppressive control, strongly inhibited the proliferation of naïve CD4+ T cells. We confirmed these data using STI571 cost 3H-thymidine incorporation assays and obtained

consistent results 27. Taken together, our studies show that we had established Th17 clones derived from TILs and that possessed characteristics of the Th17 lineage. Recent studies in humans and mice have shown that Th17 cells retain greater developmental plasticity than other types of T-cell lineages 7, 18–20. In order to maintain the cell line stability and to obtain the quantities of Th17 cells needed for future studies, we attempted to expand these Th17 clones in vitro with a standard protocol, using irradiated allogeneic PBMCs in the presence of soluble OKT3 (100 ng/mL) and IL-2. This strategy has been successfully used to expand tumor-reactive TILs for adoptive transfer immunotherapy in cancer patients 40. After each of three expansion cycles, the expanded Th17 cells were rested for 3–5 days and then analyzed for their phenotypes. We first determined IL-17, IL-4, IFN-γ-producing

cell populations and FOXP3 expression in the Th17 cells using flow cytometric analyses. Results from a representative Th17 clone were shown in Fig. 2A. We unexpectedly found that the percentages of IL-17-producing cells markedly dropped following each unbiased expansion, from over 95% before expansion (E0) to only 60% after the third expansion (E3). In contrast, the percentages of IFN-γ-producing and FOXP3+ cells were significantly Carbohydrate increased in the Th17 clones after three rounds of expansion, from 3.7 to over 60% and from 2 to 57%, respectively (Fig. 2A). Furthermore, increased proportions of IL-17+IFN-γ+ and IL-17+FOXP3+ double-positive cell populations were observed following expansion (40 and 42%, respectively, after the third round of expansion) (Fig. 2A). In addition, the percentages of IL-4-producing T-cell populations were low (<2%) in all expanded Th17 clones, and this did not change with the expansion. In addition, we obtained similar results from the other Th17 clones shown in Supporting Information Fig. 1. Notably, these expanded Th17 clones (E1–E3) maintained the same TCR-Vβ gene expression patterns as did the original Th17 clones (E0) (Fig. 1B and data not shown), suggesting the preservation of homogeneous clonality with progressive expansion.

To verify the role of mTOR Ceritinib chemical structure activation in Cd-induced neurotoxicity, mice also received a subacute regimen of intraperitoneally administered Cd

(1 mg/kg) with/without rapamycin (7.5 mg/kg) for 11 days. Chronic exposure of mice to Cd induced brain damage or neuronal cell death, due to ROS induction. Co-administration of NAC significantly reduced Cd levels in the plasma and brain of the animals. NAC prevented Cd-induced ROS and significantly attenuated Cd-induced brain damage or neuronal cell death. The protective effect of NAC was mediated, at least partially, by elevating the activities of Cu/Zn-superoxide dismutase, catalase and glutathione peroxidase, as well as the level of glutathione in the brain. Furthermore, Cd-induced activation of Akt/mTOR pathway in the brain was also inhibited by NAC. Rapamycin in vitro and in vivo protected against Cd-induced neurotoxicity. NAC protects against Cd-induced neuronal apoptosis in mouse brain partially by inhibiting ROS-dependent activation of Akt/mTOR pathway. The findings highlight that NAC may be exploited selleck products for prevention and treatment of Cd-induced neurodegenerative diseases. “
“Recent studies have indicated that bone marrow stromal cells (BMSC) may improve neurological function when transplanted into an animal model of CNS disorders, including cerebral infarct. However, there are few studies that evaluate the therapeutic benefits of intracerebral and intravenous BMSC transplantation

for cerebral infarct. This study was aimed to clarify the favorable route of cell delivery for cerebral infarct in rats. The rats were subjected to permanent middle cerebral artery occlusion.

The BMSC were labeled with near infrared (NIR)-emitting quantum dots and were transplanted stereotactically (1 × 106 cells) or intravenously (3 × 106 cells) at 7 days after the insult. Using in vivo NIR fluorescence imaging technique, CYTH4 the behaviors of BMSC were serially visualized during 4 weeks after transplantation. Motor function was also assessed. Immunohistochemistry was performed to evaluate the fate of the engrafted BMSC. Intracerebral, but not intravenous, transplantation of BMSC significantly enhanced functional recovery. In vivo NIR fluorescence imaging could clearly visualize their migration toward the cerebral infarct during 4 weeks after transplantation in the intracerebral group, but not in the intravenous, group. The BMSC were widely distributed in the ischemic brain and some of them expressed neural cell markers in the intracerebral group, but not in the intravenous group. These findings strongly suggest that intravenous administration of BMSC has limited effectiveness at clinically relevant timing and intracerebral administration should be chosen for patients with ischemic stroke, although further studies would be warranted to establish the treatment protocol. “
“We report a case of neuromyelitis optica (NMO) with an unusual pattern of remyelination in the spinal cord.

3e). At all the doses tested, there was no significant difference in IL-2 production by T cells activated by SD-4+/+ versus SD-4−/− selleck inhibitor DC. Altogether, SD-4 deletion had no impact on T-cell responses in the absence of accessory signals delivered by DC, but it augmented the DC-induced response (enhanced co-stimulatory signals resulting from lack of the inhibitory function

of DC-HIL/SD-4 between APC and T cells). Since SD-4−/− T cells were hyper-reactive to allo-antigen in the mixed lymphocyte reaction (Fig. 3a), we examined their effect on acute GVHD (Fig. 4). BALB/c mice were γ-irradiated at a sub-lethal dose and then infused with T-cell-depleted allogeneic BM cells (from C57BL/6 mice) with or without CD3+ T cells isolated from KO or WT mice. Body weight was noted weekly and survival was noted daily through to day 100. All mice lost about 30% of initial body weight within a week after BM transplantation,

but recovered some weight during the 2nd week. Thereafter, differentially treated mice displayed disparate Sotrastaurin mouse outcomes (Fig. 4a). Mice that received BM cells alone completely recovered their weight 3 weeks post-BM transplantation and survived for at least 100 days. Mice that received BM cells plus SD-4+/+ T cells partially recovered their weight, with 50% dying by day 32, and the rest survived for at least 100 days (Fig. 4b). By contrast, mice that received BM cells plus SD-4−/− T cells lost weight progressively (up to 40%) due to severe diarrhoea, with 50% dying by day 14, and all dead by day 32. We also examined proliferation of infused T cells in recipients, by measuring the number of donor-derived T cells (H-2Kb+) in spleen and liver of mice at day 5 post-BM transplantation (Fig. 4c,d). In spleen (Fig. 4c), there was twofold to threefold greater CD4+ and CD8+ SD-4−/− T cells than SD-4+/+ T cells, and also more CD69+ (activated) cells than in recipients of SD-4+/+ T cells. Similar results were observed in liver, which is another major target of acute GVHD (Fig. 4d).[1] These results indicate that infusion of T cells devoid

of SD-4 worsens morbidity and mortality of acute GVHD, most likely through hyper-reactivity to allo-antigen. Because donor-derived Treg cells are known to play a pivotal Fluorometholone Acetate role in preventing GVHD induced by co-injection of BM cells and T cells isolated from C57BL/6 mice into total body γ-irradiated BALB/c mice,[24] we studied the influence of SD-4 deletion on the T-cell-suppressive activity of Treg. We examined expression of SD-4 on conventional CD4+ Foxp3− T cells (Tconv) versus CD4+ Foxp3+ Treg cells (Fig. 5). The Tconv and Treg cells freshly isolated from naive WT mice represented 90% and 10%, respectively, and neither expressed SD-4. In contrast, PD-1 was expressed by a minuscule fraction of Tconv cells (4·6%) and by some Treg cells (22% of Foxp3+ cells) (Fig. 5a). The Tconv and Treg cells were activated by culture for 2 days with immobilized anti-CD3/CD28 antibody.

The colour reaction was stopped after 30 min and optical density was measured at 450 nm using an MRX Revelation plate reader from Dynex Technologies (Chantilly,

VA, USA). C-peptide was measured at (NLMDRL) 6 min after stimulation with 1 mg glucagon administered intravenously, as described previously [32]. All results for T cell and C-peptide are summarized as the mean, and measures of variability are reported as standard error (s.e.). Linear regression analysis was used to determine the best-fitted line, and an analysis of covariance was used to compare slopes between groups over the entire study. selleck chemicals llc Two-tailed Mann–Whitney U-tests were used to compare results at individual time-points between the treatment

groups. Two-tailed Wilcoxon matched-pairs signed-rank tests were used to compare results between individual time-points within the treatment groups. Demographic data, islet autoantibody and T cell responses to tetanus toxoid from patients treated with rosiglitazone and glyburide are shown in Table 1. No significant differences were observed in age, sex, race, body mass index (BMI), islet autoantibodies, tetanus responses or time since diagnosis between treatment groups at baseline or 36 months (Table 1). Islet-specific T cell responses in both patient groups increased during the first 12 months, becoming Caspase inhibitor increased significantly (P < 0·05) compared to baseline most at 9 months of treatment for both patient groups (Fig. 1). However, beginning at 15 months, T cell responses to islet proteins in the rosiglitazone-treated patients became suppressed significantly (P < 0·03). In fact, the T cell responses

to islet proteins in the rosiglitazone-treated patients became negative at 15 months (fewer than four blot sections) and remained negative throughout follow-up (Fig. 1). In contrast, the T cell responses to islet proteins in the glyburide patients remained positive throughout the study (Fig. 1). Mean stimulated C-peptide responses for both glyburide- and rosiglitazone-treated patients are shown in Fig. 2. During the first 12 months of follow-up, at the time T cell proliferation increased, the C-peptide in the glyburide-treated patients remained stable, whereas the C-peptide responses in the rosiglitazone-treated patients declined significantly (P < 0·05). However, after 12 months of follow-up, when islet-reactive T cell responses were suppressed in rosiglitazone-treated patients (Fig. 1), the C-peptide responses in the rosiglitazone-treated patients improved. In contrast, the C-peptide in the glyburide patients was observed to continue to decline throughout the study, reaching significance (P < 0·05) from baseline at 36 months (Fig. 2). Comparison of the glucagon-stimulated C-peptide responses for the rosiglitazone- and glyburide-treated patients demonstrated significant differences (P < 0·05) beginning at 27 months (Fig. 2).