apyrase and Diabetes-Mellitus--Type-1

Diabetes mellitus encompasses two distinct disease processes: autoimmune Type 1 (T1D) and nonimmune Type 2 (T2D) diabetes. Despite the disparate aetiologies, the disease phenotype of hyperglycemia and the associated complications are similar. In this paper, we discuss the role of the CD39-adenosinergic axis in the pathogenesis of both T1D and T2D, with particular emphasis on the role of CD39 and CD73.

Topics: Adenosine; Animals; Antigens, CD; Apyrase; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Humans; Pancreas; Signal Transduction

Type 1 diabetes (T1D) and coeliac disease are both characterized by an autoimmune feature. As T1D and coeliac disease share the same risk genes, patients risk subsequently developing the other disease. This study aimed to investigate the expression of T helper (Th), T cytotoxic (Tc) and regulatory T cells (Treg ) in T1D and/or coeliac disease children in comparison to healthy children. Subgroups of T cells (Th : CD4(+) or Tc : CD8(+) ); naive (CD27(+) CD28(+) CD45RA(+) CCR7(+) ), central memory (CD27(+) CD28(+) CD45RA(-) CCR7(+) ), effector memory (early differentiated; CD27(+) CD28(+) CD45RA(-) CCR7(-) and late differentiated; CD27(-) CD28(-) CD45RA(-) CCR7(-) ), terminally differentiated effector cells (TEMRA; CD27(-) CD28(-) CD45RA(+) CCR7(-) ) and Treg (CD4(+) CD25(+) FOXP3(+) CD127(-) ) cells, and their expression of CD39, CD45RA, CD101 and CD129, were studied by flow cytometry in T1D and/or coeliac disease children or without any of these diseases (reference group). Children diagnosed with both T1D and coeliac disease showed a higher percentage of TEMRA CD4(+) cells (P < 0·05), but lower percentages of both early and late effector memory CD8(+) cells (P < 0·05) compared to references. Children with exclusively T1D had lower median fluorescence intensity (MFI) of forkhead box protein 3 (FoxP3) (P < 0·05) and also a lower percentage of CD39(+) and CD45RA(+) within the Treg population (CD4(+) CD25(+) FOXP3(+) CD127(-) ) (P < 0·05). Children with exclusively coeliac disease had a higher MFI of CD101 (P < 0·01), as well as a higher percentage of CD129(+) (P < 0·05), in the CD4(+) CD25(hi) lymphocyte population, compared to references. In conclusion, children with combined T1D and coeliac disease have a higher percentage of differentiated CD4(+) cells compared to CD8(+) cells. T1D children show signs of low CD39(+) /CD45RA(+) Treg cells that may indicate loss of suppressive function. Conversely, children with coeliac disease show signs of CD101(+) /CD129(+) Treg cells that may indicate suppressor activity.

Topics: Adolescent; Antigens, CD; Apyrase; CD8-Positive T-Lymphocytes; Celiac Disease; Child; Diabetes Mellitus, Type 1; Female; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Leukocyte Common Antigens; Male; Membrane Glycoproteins; Receptors, Interleukin-9; T-Lymphocytes, Regulatory

Diabetes mellitus (DM) is a metabolic disorder characterized by a chronic hyperglycemia state, increased oxidative stress parameters, and inflammatory processes.. To evaluate the effect of caffeic acid (CA) on ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) and adenosine deaminase (ADA) enzymatic activity and expression of the A2A receptor of the purinergic system, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymatic activity and expression of the α7nAChR receptor of the cholinergic system as well as inflammatory and oxidative parameters in diabetic rats.. Diabetes was induced by a single dose intraperitoneally of streptozotocin (STZ, 55 mg/kg). Animals were divided into six groups (n = 10): control/oil; control/CA 10 mg/kg; control/CA 50 mg/kg; diabetic/oil; diabetic/CA 10 mg/kg; and diabetic/CA 50 mg/kg treated for thirty days by gavage.. CA treatment reduced ATP and ADP hydrolysis (lymphocytes) and ATP levels (serum), and reversed the increase in ADA and AChE (lymphocytes), BuChE (serum), and myeloperoxidase (MPO, plasma) activities in diabetic rats. CA treatment did not attenuate the increase in IL-1β and IL-6 gene expression (lymphocytes) in the diabetic state; however, it increased IL-10 and A2A gene expression, regardless of the animals' condition (healthy or diabetic), and α7nAChR gene expression. Additionally, CA attenuated the increase in oxidative stress markers and reversed the decrease in antioxidant parameters of diabetic animals.. Overall, our findings indicated that CA treatment positively modulated purinergic and cholinergic enzyme activities and receptor expression, and improved oxi-inflammatory parameters, thus suggesting that this phenolic acid could improve redox homeostasis dysregulation and purinergic and cholinergic signaling in the diabetic state.

Topics: Acetylcholinesterase; Adenosine Deaminase; alpha7 Nicotinic Acetylcholine Receptor; Animals; Antigens, CD; Antioxidants; Apyrase; Butyrylcholinesterase; Caffeic Acids; Cytokines; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Gene Expression Regulation; GPI-Linked Proteins; Inflammation Mediators; Lipid Peroxidation; Lymphocytes; Male; Membrane Proteins; Oxidative Stress; Peroxidase; Rats; Rats, Wistar

Type 1 diabetes (T1D) is an autoimmune disease resulting from an attack by autoreactive T lymphocytes against pancreatic islet β- cells. In recent studies, regulatory T cells (Tregs) have been implicated in the process of T1D. Furthermore, cluster of differentiation 39 (CD39), which is involved in the suppression of inflammation, has been shown to be expressed on Tregs. However, the pathological importance of CD39 to the memory Treg population remains unclear.. This study investigated Treg subsets, focusing on resting, effector, and memory Tregs, and determined CD39 expression on Tregs. In addition, changes in Treg subsets and Treg-associated cytokine secretion after CD3/CD28 stimulation of peripheral blood mononuclear cells were evaluated in diabetic patients and healthy controls. The suppressive function of Tregs was measured using the mixed lymphocyte reaction (MLR) test.. There was a higher percentage of memory Tregs in T1D patients than healthy controls. However, Tregs in T1D patients showed impaired suppression, with low forkhead box P3 (Foxp3) expression and low serum interleukin (IL)-10 levels. Furthermore, CD39 expression on Tregs, and on memory Tregs in particular, was lower in T1D patients than healthy controls. After stimulation, the percentage of resting Tregs was decreased and that of effector/memory Tregs was increased in both healthy controls and T1D patients, but CD39 expression on effector/memory Tregs was still lower and there was no increase in IL-10 secretion in T1D patients.. The defective suppressive function of Tregs in T1D patients is due to lower expression of CD39 on memory Tregs.. 摘要: 背景 1型糖尿病是一种由自身T淋巴细胞攻击胰岛β细胞而造成的自身免疫性疾病。最近有研究表明调节性T细胞(regulatory T cells，Tregs)在1型糖尿病发病过程中发挥作用，此外也有研究证明分化抗原簇39(CD39)参与炎症抑制过程，且在Tregs上表达。然而，CD39在记忆型Treg亚群中病理学上的重要性仍未阐明。 方法 该研究将分析Tregs亚群，包括静息、效应和记忆Tregs亚群，检测CD39是否在这些Tregs亚群中表达。在患者和健康对照者中，以CD3/CD28刺激外周血单核细胞，观察Tregs亚群及Treg相关细胞因子分泌情况的变化。同时，在混合淋巴细胞反应中检测患者和健康对照Treg的抑制功能。 结果 虽然该研究发现1型糖尿病患者中记忆型Treg亚群比例高于健康对照，但是，1型糖尿病患者Treg的抑制功能受损，同时其Foxp3的表达降低及血清中IL-10分泌水平降低，并且CD39在Tregs中的表达，尤其是在记忆Tregs中的表达低于健康对照。在CD3/CD28磁珠刺激后，不管是患者还是健康对照，静息Treg亚群的比例降低，效应/记忆Treg亚群升高。但是1型糖尿病患者中，PBMCs经过刺激后，CD39在效应/记忆Treg亚群中仍然维持低表达，而IL-10的分泌也并没有上升。 结论 1型糖尿病患者中Treg抑制功能的受损是由于CD39在记忆Treg亚群中的表达降低所致。.

Topics: Adult; Apyrase; Biomarkers; Blood Glucose; Case-Control Studies; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Female; Follow-Up Studies; Forkhead Transcription Factors; Glycated Hemoglobin; Humans; Immunologic Memory; Interleukin-10; Leukocytes, Mononuclear; Male; Middle Aged; Prognosis; T-Lymphocytes, Regulatory

Polyclonal T regulatory cells (Treg - CD4

Topics: Animals; Antigens, CD; Apyrase; Cell Proliferation; Cell Separation; Coculture Techniques; Dendritic Cells; Diabetes Mellitus, Type 1; Disease Models, Animal; Gene Expression; Humans; Insulin; Interleukin-10; Interleukin-2; Mice; Mice, Inbred NOD; Primary Cell Culture; Programmed Cell Death 1 Receptor; Receptors, Antigen, T-Cell; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

We investigated the relationship between inflammation, neuronal loss, and expression of indoleamine 2, 3-dioxygenase (IDO) and quinolinic acid (QUIN) in the retina of subjects with type 1 diabetes (T1D) and type 2 diabetes (T2D) and in the retina of rats with T1D.. Retinas from T1D (n = 7), T2D (n = 13), and 20 age-matched nondiabetic human donors and from T1D (n = 3) and control rats (n = 3) were examined using immunohistochemistry for IDO, QUIN, cluster of differentiation 39 (CD39), ionized calcium-binding adaptor molecule (Iba-1, for macrophages and microglia), Vimentin (VIM; for Müller cells), neuronal nuclei (NeuN; for neurons), and UEA1 lectin (for blood vessels).. Based on morphologic criteria, CD39+/ionized calcium binding adaptor molecule 1(Iba-1+) resident microglia and CD39-/Iba-1+ bone marrow-derived macrophages were present at higher density in T1D (13% increase) and T2D (26% increase) human retinas when compared with controls. The density and brightness of IDO+ microglia were increased in both T1D and T2D human retinas. The intensity of QUIN+ expression on CD39+ microglia and VIM+ Müller cells was greatly increased in both human T1D and T2D retinas. T1D retinas showed a 63% loss of NeuN+ neurons and T2D retinas lost approximately 43% when compared with nondiabetic human retinas. Few QUIN+ microglia-like cells were seen in nondiabetic retinas, but the numbers increased 18-fold in T1D and 7-fold in T2D in the central retina. In T1D rat retinas, the density of IDO+ microglia increased 2.8-fold and brightness increased 2.1-fold when compared with controls.. Our findings suggest that IDO and QUIN expression in the retinas of diabetic rats and humans could contribute to the neuronal degeneration that is characteristic of diabetic retinopathy.

Topics: Aged; Animals; Antigens, CD; Antigens, Nuclear; Apyrase; Biomarkers; Calcium-Binding Proteins; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; DNA-Binding Proteins; Ependymoglial Cells; Female; Fluorescent Antibody Technique, Indirect; Humans; Indoleamine-Pyrrole 2,3,-Dioxygenase; Male; Microfilament Proteins; Microglia; Microscopy, Confocal; Middle Aged; Nerve Tissue Proteins; Quinolinic Acid; Rats; Rats, Sprague-Dawley; Retina; Vimentin

The authors used histochemical analysis to determine whether increased vascular endothelial growth factor (VEGF) immunoreactivity in diabetic retinal vessels is related to increased vascular permeability, as indicated by human serum albumin (HSA) immunostaining, or to presumed retinal hypoxia as demonstrated by decreased vascularity. A correlation between VEGF and HSA in cryosections with angiopathic changes in the adenosine diphosphatase (ADPase) flat-embedded fellow retinas was sought. Because VEGF is a heparin-binding protein, the relation between VEGF and heparan sulfate proteoglycan (HSPG) immunoreactivities was also investigated.. Cryopreserved eyes from 18 diabetic and 9 nondiabetic subjects removed after death were sectioned and immunohistochemical analysis was performed with antibodies against VEGF, HSA, HSPG, vWf (von Willebrand factor), and collagen IV. The fellow retinas were prepared by our ADPase flat-embedding technique to determine the degree of diabetic retinopathy. The number of positive vessels for each antibody and antibody localizations were determined by light microscopy.. The average number of VEGF-stained vessels in diabetic retinas was significantly higher than in nondiabetic retinas (P = 0.04). In diabetic retinas, there was a positive correlation between the distribution of VEGF-positive vessels and the distribution of HSA- and HSPG-positive vessels. No such correlation was observed in nondiabetic eyes. In many cases, HSPG immunoreactivity appeared colocalized with VEGF immunoreactivity, suggesting VEGF binding to HSPG. The comparison with the ADPase flat-embedded fellow retinas suggested that increased VEGF immunoreactivity and vascular permeability may occur before morphologic changes in the vasculature.. Vascular endothelial growth factor immunoreactivity was correlated with increased vascular permeability to macromolecules and appears to be increased in diabetic subjects before the onset of retinopathy.

Topics: Adult; Aged; Aged, 80 and over; Apyrase; Capillary Permeability; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Endothelial Growth Factors; Heparan Sulfate Proteoglycans; Humans; Immunoenzyme Techniques; Lymphokines; Middle Aged; Retinal Vessels; Serum Albumin; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

While it is known that platelets from diabetic patients bind more fibrinogen than do platelets from normal subjects, there has been no study comparing this phenomenon in platelets from nonretinopathic and retinopathic patients. We have made such a comparison and have found the following. In agreement with previous reports, platelets from nonretinopathic diabetic patients bind abnormally high amounts of fibrinogen. No differences in the amount of fibrinogen bound to platelets (stimulated by collagen or thrombin) were found when data from nonretinopathic and retinopathic patients were compared. However, while aspirin (an inhibitor of thromboxane synthesis) reduced the abnormally high fibrinogen binding of platelets from nonretinopathic patients to normal control levels, it did not normalize the high fibrinogen binding of platelets from retinopathic diabetic patients. The combination of aspirin plus apyrase (an ADP scavenger) almost suppressed fibrinogen binding and aggregation of platelets from normal or nonretinopathic diabetic subjects, whereas it had a somewhat lesser effect on binding and aggregation of platelets from retinopathic subjects. By using a monoclonal antibody (B59.2) to the platelet receptor for fibrinogen, we determined that this receptor was the same in platelets from normal as well as nonretinopathic diabetic subjects and that this antibody could suppress the binding of fibrinogen and the aggregation of platelets from both types of patients just as it did in platelets from normal subjects. Thus, our data indicate that, while platelets from both retinopathic and nonretinopathic patients are hyperaggregable and show abnormally high binding of fibrinogen, they differ in that these abnormalities can be normalized in platelets from nonretinopathic patients by suppressing prostaglandin/thromboxane formation and scavenging ADP, but not in those from retinopathic patients.

Topics: Adolescent; Adult; Apyrase; Aspirin; Blood Platelets; Diabetes Mellitus, Type 1; Diabetic Retinopathy; Female; Fibrinogen; Humans; Male; Platelet Aggregation