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M Previti

Publications and source records attributed to M Previti.

11 recordsLinked to original sources

Escape from the acute Wolff-Chaikoff effect is associated with a decrease in thyroid sodium/iodide symporter messenger ribonucleic acid and protein.

In 1948, Wolff and Chaikoff reported that organic binding of iodide in the thyroid was decreased when plasma iodide levels were elevated (acute Wolff-Chaikoff effect), and that adaptation or escape from the acute effect occurred in approximately 2 days, in the presence of continued high plasma iodide concentrations. We later demonstrated that the escape is attributable to a decrease in iodide transport into the thyroid, lowering the intrathyroidal iodine content below a critical inhibitory threshold and allowing organification of iodide to resume. We have now measured the rat thyroid sodium/iodide symporter (NIS) messenger RNA (mRNA) and protein levels, in response to both chronic and acute iodide excess, in an attempt to determine the mechanism responsible for the decreased iodide transport. Rats were given 0.05% NaI in their drinking water for 1 and 6 days in the chronic experiments, and a single 2000-microg dose of NaI i.p. in the acute experiments. Serum was collected for iodine and hormone measurements, and thyroids were frozen for subsequent measurement of NIS, TSH receptor, thyroid peroxidase (TPO), thyroglobulin, and cyclophilin mRNAs (by Northern blotting) as well as NIS protein (by Western blotting). Serum T4 and T3 concentrations were significantly decreased at 1 day in the chronic experiments and returned to normal at 6 days, and were unchanged in the acute experiments. Serum TSH levels were unchanged in both paradigms. Both NIS mRNA and protein were decreased at 1 and 6 days after chronic iodide ingestion. NIS mRNA was decreased at 6 and 24 h after acute iodide administration, whereas NIS protein was decreased only at 24 h. TPO mRNA was decreased at 6 days of chronic iodide ingestion and 24 h after acute iodide administration. There were no iodide-induced changes in TSH receptor and thyroglobulin mRNAs. These data suggest that iodide administration decreases both NIS mRNA and protein expression, by a mechanism that is likely to be, at least in part, transcriptional. Our findings support the hypothesis that the escape from the acute Wolff-Chaikoff effect is caused by a decrease in NIS, with a resultant decreased iodide transport into the thyroid. The observed decrease in TPO mRNA may contribute to the iodine-induced hypothyroidism that is common in patients with Hashimoto's thyroiditis.

Animals↗

The GM2-1 ganglioside islet autoantigen in insulin-dependent diabetes mellitus is expressed in secretory granules and is not beta-cell specific.

The pancreatic islet monosialo-ganglioside (GM2-1), an autoantigen in insulin-dependent diabetes mellitus (IDDM) recently shown to be the target of autoantibodies associated with diabetes development in relatives of IDDM patients, is islet specific within the pancreas, and its expression is metabolically regulatable. In the present study we sought to establish 1) whether GM2-1 is beta-cell specific, and 2) its intracellular localization. To this end, we analyzed the pattern of ganglioside expression in highly purified beta- and non-beta-cells isolated from rat islets. In addition, ganglioside levels were determined in subcellular fractions of a rat beta-cell line (INS). No qualitative or quantitative difference was found in the pattern of ganglioside expression between beta and non-beta rat islet cells, with GM3, GM2-1, and GD3 gangliosides expressed in both cell populations. Within INS cells, GM2-1 ganglioside was expressed in the fraction containing secretory granules and, to a lesser extent, in plasma membranes; GM3 was expressed in secretory granules, whereas GD3 was found only in plasma membranes. These data indicate that the GM2-1 autoantigen is not beta-cell specific within the islets, in accordance with the observation that this molecule is a target of islet cell autoantibodies that bind to the whole pancreatic islet. Interestingly, this autoantigen is present in secretory granules similarly to other autoantigens in IDDM (insulin, carboxypeptidase H, 38-kDa protein, etc.), suggesting that the autoimmunity to the components of this organelle may be central to the pathogenesis of the disease.

Animals↗

Interferon-alpha therapy may induce insulin autoantibody development in patients with chronic viral hepatitis.

Development of type 1 insulin-dependent diabetes mellitus has been recently reported in patients who underwent interferon-alpha (IFN-alpha) therapy because of chronic viral hepatitis. Furthermore IFN-alpha seems to be involved in the immunological events that lead to beta-cell destruction and development of type 1 diabetes. To evaluate whether IFN-alpha treatment could elicit an autoimmune response against beta-cell antigens, we determined the occurrence of islet cell antibodies and insulin autoantibodies in the sera of 60 patients with HCV- or HBV-related chronic hepatitis who had been treated with IFN-alpha for 6 or 12 months. The presence of antibodies against thyroglobulin, thyroid microsomal antigen, gastric parietal cells, and non-organ-specific antigens was also investigated. Insulin autoantibody positivity was observed in 2/60 (3.3%), 8/60 (13.3%), and 4/30 (13.3%) patients, before IFN-alpha treatment, and after 6 months and 12 months of therapy, respectively. None of the studied patients developed islet cell antibodies or type 1 diabetes. Before IFN-alpha therapy four patients showed thyroid autoantibodies and four others developed antibodies against thyroglobulin and/or thyroid microsomal antigen during the treatment. Coexistence of insulin autoantibodies and thyroid autoantibodies was observed in only two patients. Our results showed that IFN-alpha therapy in patients with chronic viral hepatitis is capable of inducing development of autoantibodies against insulin. This event seems to be not related to other autoimmune disorders.

Adolescent↗

Beta-cell markers and autoantigen expression by a human insulinoma cell line: similarities to native beta cells.

In the present study we have evaluated the expression of different beta-cell markers, islet molecules and auto-antigens relevant in diabetes autoimmunity by a human insulinoma cell line (CM) in order to define its similarities with native beta cells and to discover whether it could be considered as a model for studies on immunological aspects of Type 1 diabetes. First, the positivity of the CM cell line for known markers of neuroendocrine derivation was determined by means of immunocytochemical analysis using different anti-islet monoclonal antibodies including A2B5 and 3G5 reacting with islet gangliosides, and HISL19 binding to an islet glycoprotein. Secondly, the expression and characteristics of glutamic acid decarboxylase (GAD) and of GM2-1 ganglioside, both known to be islet autoantigens in diabetes autoimmunity and expressed by human native beta cells, were investigated in the CM cell line. The pattern of ganglioside expression in comparison to that of native beta cells was also evaluated. Thirdly, the binding of diabetic sera to CM cells reacting with islet cytoplasmic antigens (ICA) was studied by immunohistochemistry. The results of this study showed that beta cell markers identified by anti-islet monoclonal antibodies A2B5, 3G5 and HISL-19 are expressed by CM cells; similarly, islet molecules such as GAD and GM2-1 ganglioside are present and possess similar characteristics to those found in native beta cells; the pattern of expression of other gangliosides by CM cells is also identical to human pancreatic islets; beta cell autoantigen(s) reacting with antibodies present in islet cell antibodies (ICA) positive diabetic sera identified by ICA binding are also detectable in this insulinoma cell line. We conclude that CM cells show close similarities to native beta cells with respect to the expression of neuro-endocrine markers, relevant beta cell autoantigens in Type 1 diabetes (GAD, GM2-1, ICA antigen), and other gangliosides. Therefore, this insulinoma cell line may be considered as an ideal model for studies aimed at investigating autoimmune phenomena occurring in Type 1 diabetes.

Autoantigens↗

Autoimmunity to the GM2-1 islet ganglioside before and at the onset of type I diabetes.

Recently, the GM2-1 pancreatic islet ganglioside, proposed as a potential autoantigen in type I diabetes autoimmunity, has been biochemically characterized and found to be a novel ganglioside structure. In the present study, we aimed to determine whether an autoimmune response toward this novel islet molecule is 1) present in type I diabetes and is specifically directed against this molecule and not to gangliosides in general and 2) predictive of disease in high-risk subjects. To this end, the following patients have been studied: 1) 24 newly diagnosed type I diabetic subjects, 20 islet cell autoantibody (ICA)-negative first-degree relatives of type I diabetic subjects, and 25 age-matched normal control individuals; and 2) 31 prospectively evaluated ICA+ first-degree relatives of type I diabetic subjects who were followed for up to 10 years, during which 14 of them developed type I diabetes. A direct assay for autoantibodies to GM2-1 and to other pancreatic gangliosides (GM3, GD3, GD1a) was developed using an indirect immunoperoxidase technique performed directly on thin layer chromatography plates. Anti-GM2-1 autoantibodies (all belonging to the IgG class) were expressed in a high percentage of newly diagnosed type I diabetic subjects (71%), while no significant difference was found in the expression of antibodies directed against other pancreatic gangliosides (GM3, GD3, GD1a) among the different groups studied. Anti-GM2-1 autoantibodies were also present in ICA+ relatives (64%) (P < 0.001 vs. control subjects and ICA-relatives): in this group, life table analysis of progression to diabetes showed that anti-GM2-1 autoantibodies were significantly (P < 0.001) associated with disease, occurring in all relatives developing type I diabetes within 5 years and thus identifying a cohort of ICA+ subjects with markedly increased diabetes risk.

Autoantibodies↗

GM2-1 pancreatic islet ganglioside: identification and characterization of a novel islet-specific molecule.

Recent studies have indicated that GM2-1, a pancreatic islet monosialo-ganglioside, is an islet-specific component whose expression is metabolically regulable and represents one of the target antigens of cytoplasmic islet cell antibodies. In the present study we aimed to biochemically characterize this molecule using a panel of biochemical techniques including gas chromatography, thin layer chromatography, enzymatic digestion and mass spectrometry. GM2-1 ganglioside was extracted from human pancreas and purified by thin-layer chromatography. Fatty acids in the ceramide (the hydrophobic portion of the molecule), identified by gas chromatography ranged from C16:1 to C24:1. The oligosaccharide chain was enzymatically digested by the sequential application of various exoglycosidases (neuraminidase followed by beta-galactosidase, followed by beta-hexosaminidase) and characterized by gas chromatography identification of the liberated sugars. The following structure was deducted from enzymatic studies and confirmed by mass spectrometry analysis: N-acetyl neuraminic acid-galactose-galactosamine-galactosamine-glucose-ceramide. This is a novel ganglioside structure, not yet described, which shares characteristics with a neuronal glycolipid autoantigen: the LM1 ganglioside. Both GM2-1 and LM1 have a single sialic acid residue in the terminal position, the same migration position on thin layer chromatography and the same number of carbohydrate moieties. In conclusion, we have characterized a novel islet-specific ganglioside molecule with unusual characteristics, such as the terminal sialic acid and the galactosamine residues, which may facilitate both its antigenicity and its involvement in beta-cell autoimmunity.

Carbohydrate Sequence↗

Gastric mucosal mast cells in atopic subjects.

Intragastral allergen provocation under endoscopic control (IPEC) allows direct observation of gastric mucosa reactions after contact with inhalant allergens that reach the stomach. We selected patients with proved atopy to Parietaria but without clinical and endoscopic signs of gastric disease, and we tested them with the specific inhalant allergen during IPEC, recording gastric macroscopic reaction and mucosal mast-cell changes in biopsy specimens. All atopic patients showed visible changes in gastric mucosa quantified as IPEC score. Mast-cell numbers detected in atopic patients (135.4 +/- 102.6/mm2 of stromal area) were significantly higher than in nonatopic subjects (59.8 +/- 25.4/mm2; P < 0.03) and were positively correlated to atopic IPEC score (P < 0.01). In addition, 6/12 atopics who had both higher mast-cell counts and IPEC score showed an intraepithelial distribution of gastric mast cells which displayed ultrastructural features of partial degranulation. It is likely that changes observed in our patients with allergy to Parietaria reflect a subclinical activation of mast cells in the gastric mucosa.

Adolescent↗

High serum levels of soluble CD8 in insulin-dependent diabetes.

In type 1 (insulin-dependent) diabetes mellitus (IDDM) CD8+ T cells represent the majority of lymphocytes which infiltrate the pancreatic islets during beta cell destruction. Soluble CD8 antigen (sCD8) has been shown to correlate with CD8 cell subset activation. In this study we measured by ELISA sCD8 levels in sera from: 33 newly diagnosed IDDM patients; 29 type 1 diabetics with duration of disease more than 1 year; 37 healthy siblings of IDDM patients; 19 healthy controls. Sera from both groups of IDDM patients and from healthy siblings exhibited soluble CD8 mean levels significantly higher than controls (P = 0.0001, P < 0.003, P < 0.03 respectively). Soluble CD8 levels above the normal range (mean +/- 2 s.d. of controls) were found in a percentage of newly diagnosed subjects (54.5%) significantly higher than in subjects with a long-standing duration of disease (6.9%, P < 0.0005) and healthy siblings (16.2%, P < 0.002). Our results suggest that the raised levels of soluble CD8 near to diabetes onset may indicate the activation of CD8+ T cells probably responsible for the autoimmune beta cell destruction.

Adolescent↗

Rat pancreatic ganglioside expression: differences between a model of autoimmune islet B cell destruction and a normal strain.

Islet cell antibodies (ICA) bind antigens expressed in both human and rat pancreatic islets. Biochemical studies have shown that an ICA-autoantigen has the properties of a monosialo-ganglioside migrating between GM2 and GM1 standards (GM2-1). We therefore aimed to isolate and characterize gangliosides from whole pancreas and isolated islets of bio breeding diabetes-prone (BB-DP), bio breeding diabetes-resistant (BB-DR), and Wistar Furth (WF) rat strains. Gangliosides were characterized by TLC, HPLC, diode array analysis, and ganglioside-specific staining. ICA binding was studied by indirect immunostaining. The GM2-1 fraction was present in BB-DP, BB-DR, and WF rat pancreases (11, 17, and 9.5%, respectively, of total ganglioside content). Substantial differences were found in other fractions: in BB-DP pancreas, in addition to GM2-1, the main fractions were GM3 (49%), GD1a (12%), GT1b (5%), and a ganglioside migrating between GM1 and GD3 standards (23%), while in BB-DR pancreas the above components were 71, 5.5, 2, and 4.5%, respectively; in WF pancreas, the main fractions were GM3, GD3, GD1a, GT1b and a trisialoganglioside (GT*) migrating above the GT1b standard (42.7, 7, 20.2, 13.8, and 6.8, respectively). A different pattern of ganglioside expression was found in isolated islets of BB-DP, BB-DR, and WF rats: the GM2-1 fraction represented, respectively, 29.1, 30.4, and 31.6% of total ganglioside content; GM3 51.1, 66, and 68.4%. A fraction migrating between GM1 and GD3 standards was present only in BB-DP and BB-DR islets (19.8 and 3.6%, respectively). ICA-positive human sera reacted with pancreas of all rat strains studied, with similar end-point titers. In conclusion, (1) the GM2-1 ganglioside, in the same way as a putative target antigen of ICA, is equally expressed in BB-DP, BB-DR, and WF rat pancreata; and (2) the GM1-GD3 is expressed in higher amounts in BB-DP than in BB-DR pancreas and islets and is absent in WF.

Animals↗

Pancreatic islet ganglioside expression in nonobese diabetic mice: comparison with C57BL/10 mice and changes after autoimmune beta-cell destruction.

Recent observations have shown that the presumed target antigen of cytoplasmic islet cell antibodies (ICA) has properties of a monosialo-ganglioside migrating between GM2 and GM1 standards (GM2-1) and that ICA binding is higher in nonobese diabetic (NOD) than in C57BL/10SnJ mouse pancreatic frozen sections. This study aimed to characterize the ganglioside expression in NOD mouse islets in comparison with the control C57BL/10SnJ strain, taking into account possible sex differences, variations with age, and changes after autoimmune beta-cell destruction. Thus, acidic glycolipid composition was analyzed 1) in isolated islets from 11-week-old female and male NOD mice and age-matched female and male C57BL/10SnJ mice, and 2) in whole pancreas of both NOD and control mouse strains at different ages (4, 8, and 18 weeks) and of female NOD mice before and after diabetes onset. The acidic glycolipid GM2-1 is expressed in isolated female NOD islets, male NOD islets, and C57BL/10SnJ mouse islets, but quantitative analysis showed an increased amount of GM2-1 in NOD vs. C57BL/10 islets. GM3 is a ganglioside fraction expressed in female and male NOD mice and not in the C57BL/10 strain, whereas GD3 characterizes the C57BL/10 strain islets. GM2-1 is the sole ganglioside fraction in the whole pancreas to clearly decrease with age in the NOD mouse, and diabetes onset in this strain is associated with a significant decrease in the expression of this component as well as of GM3, whereas other pancreatic ganglioside (GD3, GD1a, and GT1b) levels did not significantly decrease; no age-related ganglioside change was observed in the C57BL/10SnJ mouse. Interestingly, the observed increased ICA binding in NOD islets is paralleled by the increased expression of GM2-1 islet ganglioside, and beta-cell destruction in NOD mice is associated with a significant decrease in the amount of this ganglioside in the pancreas.

Age Factors↗

Advance directives.

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Advance Directives↗