Masako Waguri

Mononuclear cell infiltration and its relation to the expression of major histocompatibility complex antigens and adhesion molecules in pancreas biopsy specimens from newly diagnosed insulin-dependent diabetes mellitus patients.

We examined pancreas biopsy specimens from 18 newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients to elucidate the mechanism underlying beta cell destruction. Pancreas islets were seen in all patients and insulitis in eight patients. Infiltrating mononuclear cells consisted of CD4+T, CD8+T, B lymphocytes, and macrophages. Among them, CD8+T lymphocytes were predominant and macrophages followed. The expression of MHC class I antigens was increased in islet and endothelial cells in nine patients. MHC class II expression was increased in endothelial cells of the same patients. The expression of intercellular adhesion molecule-1 was increased in endothelial cells in two of the nine patients with MHC hyperexpression; in one of them, lymphocyte function-associated antigen-3 expression was also increased. Out of the eight patients with insulitis, seven showed MHC class I hyper-expression, whereas 2 of the 10 patients without insulitis showed the phenomenon (P < 0.05). The relation between insulitis and the hyperexpression of adhesion molecules was not evident. In conclusion, we revealed the close relation between CD8+T lymphocyte-predominant insulitis and MHC class I hyperexpression in islet cells. This suggests that infiltrating CD8+T lymphocytes recognize islet autoantigens in association with increased MHC class I molecules and act as major effector cells in autoimmune response against islet cells in IDDM pancreases. The role of adhesion molecules in the pathogenesis of IDDM still remains to be elucidated.

Demonstration of Two Different Processes of β-Cell Regeneration in a New Diabetic Mouse Model Induced by Selective Perfusion of Alloxan

To clarify the regeneration process of pancreatic β-cells, we established a new mouse model of diabetes induced by selective perfusion of alloxan after clamping the superior mesenteric artery. In this model, diabetes could be induced by the destruction of β-cells in alloxan-perfused segments, while β-cells in nonperfused segments were spared. Intraperitoneal glucose tolerance tests showed glucose intolerance, which gradually ameliorated and was completely normalized in 1 year with a concomitant increase of insulin content in the pancreas. Histological examination showed neoislet formation in the alloxan-perfused segment and the proliferation of spared β-cells in the nonperfused segment. In the alloxan-perfused segment, despite a marked reduction of islets in size and number at an early stage, both the number of islets, including islet-like cell clusters (ICCs), and the relative islet area significantly increased at a later stage. Increased single β-cells and ICCs were located in close contact with duct cell lining, suggesting that they differentiated from duct cells and that such extra-islet precursor cells may be important for β-cell regeneration in β-cell–depleted segment. In addition to β-cells, some nonhormone cells in ICCs were positive for nuclear insulin promoter factor 1, which indicated that most, if not all, nonhormone cells positive for this factor were β-cell precursors. In the nonperfused segment, the islet area increased significantly, and the highest 5-bromo-2-deoxyuridine–labeling index in β-cells was observed at day 5, while the number of islets did not increase significantly. This indicated that the regeneration of islet endocrine cells occurs mostly through the proliferation of preexisting intra-islet β-cells in the nonperfused segment. In conclusion, the regeneration process of β-cells varied by circumstance. Our mouse model is useful for studying the mechanism of regeneration, since differentiation and proliferation could be analyzed separately in one pancreas.

Occurrence of IDDM during interferon therapy for chronic viral hepatitis

We report a case of IDDM which occurred during interferon therapy for chronic hepatitis. A 31-year-old man intermittently received 2.5 x 10(8) units of alpha-IFN and 1 x 10(8) units of beta-IFN for treatment of chronic viral hepatitis type B. Four years after the beginning of IFN therapy, he acutely developed moderate hyperglycemia and severe ketonuria with positive islet cell antibody, and then 28 units/day of insulin injection was started. After the start of insulin therapy, there was a remission period for about 3 years but insulin-dependency recurred thereafter. The clinical course of this case indicates that IFN therapy precedes IDDM. During and after IFN therapy we should consider the possibility of occurrence of IDDM as well as other autoimmune diseases and observe the clinical course carefully.

Immunological abnormalities in islets at diagnosis paralleled further deterioration of glycaemic control in patients with recent-onset Type I (insulin-dependent) diabetes mellitus

Aims/hypothesis. To determine whether the clinical heterogeneity observed in the development of Type I (insulin-dependent) diabetes mellitus correlates with immunohistochemical differences observed at diagnosis. Methods. Patients (n = 17) with recent-onset diabetes clinically considered to be insulin dependent (Type I), underwent pancreatic biopsy for immunohistological analysis. These patients were divided into two groups based on the presence or absence of islet immunological abnormalities (insulitis or hyperexpression of MHC class I antigens or both). The patients were also HLA typed and tested for islet cell antibodies and antibodies to glutamic acid decarboxylase (GAD-Ab). All patients were followed monthly for 2 years and their fasting plasma glucose, haemoglobin A1C and daily insulin doses were recorded. The clinical course of patients with islet immunological abnormalities was compared with that of patients without those abnormalities. Results. Patients with and without islet immunological abnormalities did not differ with regard to HLA type or islet cell antibodies. Antibodies to glutamic acid decarboxylase correlated with the presence of insulitis and MHC class I hyperexpression. These local immunological abnormalities were also associated with higher haemoglobin A1C values (p < 0.05) and a trend towards greater insulin requirements. Further, patients with the islet abnormalities had higher fasting plasma glucose concentrations 2 years after the biopsy than at the time of the biopsy (p < 0.05). Conclusion/interpretation. The heterogeneous clinical course observed following diagnosis in patients with Type I diabetes correlates with islet immunological abnormalities. Insulitis and hyperexpression of MHC class I correlate with deteriorating glycaemic control. [Diabetologia (1999) 42: 574–578]

Antibodies to glutamic acid decarboxylase induced by interferon-alpha therapy for chronic viral hepatitis

Dear Sir, We and others have reported cases of insulin-dependent diabetes mellitus (IDDM) that developed during interferon-alpha therapy [1, 2]. Foulis et al. [3] have shown that immunoreactive interferon-alpha was positive in insulin-secreting beta cells in IDDM patients, and, recently, the expression of mRNA of interferon-alpha in the IDDM pancreas has been demonstrated [4]. These findings indicate that interferon-alpha plays a role in the development of 1DDM. In addition to classical islet-cell antibodies, serum antibodies to glutamic acid decarboxylase (GAD) are good predictive and diagnostic markers for IDDM [5], and several simple assays for GAD antibodies have recently become available. To clarify the role of interferon-alpha in the pathogenesis of IDDM, we examined the status of GAD antibodies in a patient who developed IDDM during interferon therapy. The detailed profile and treatment of the patient were reported previously [1]. Briefly, hewas a 31-year-old man, who received a dosage of 54 x 106 units of recombinant interferon-alpha 2a (Canferon A; Takeda Chemical, Osaka, Japan) for chronic active type-B hepatitis, which was followed by the development of IDDM. Antibodies to GAD were measured by a radioimmunoassay kit using GAD purified from porcine brain as an antigen (Hoechst Japan, Tokyo, Japan) [5]. Islet-cell antibodies were detected with an indirect immtmofluorescence method and reported in Juvenile Diabetes Foundation Units (JDFU) as described previously [6]. The cutoff levels of GAD antibodies and islet-cell antibodies were 5 U/ml and 5 JDFU, respectively. The results showed that GAD antibodies and islet-cell antibodies were positive 2 weeks after the onset of overt diabetes in this patient. The antibodie titre to GAD was 622 U/ml and that of islet-cell antibodie was 80 JDFU. Antibodies to GAD and islet-cell antibodies remained positive at 328 U/ml and 40 JDFU, respectively 2.5 years after diabetes development. (Pretreatment serum was not available.) This prompted us to further investigate the status of GAD antibodies in another 40 patients [26 males, 14 females, mean age _+ SD of 47.9 -+ 10.1 years] who received recombinant interferon-alpha 2a (Canferon A, or Roferon A; La Roche, Nutley, N.J. USA) or 2b (Intron A; Schering, Kenilworth, N.J., USA) for 24 weeks with a total dosage of 114 to 845 x 106 units [368 +130 x 106 units] to treat chronic viral hepatitis. The sera were taken from the patients before and 4,12, and 24 weeks after the start of treatment. As a result, GAD antibodies newly appeared at 8 U/ml 4 weeks after the start of the interferon

Comparison of pregnancy outcomes between women with gestational diabetes and overt diabetes first diagnosed in pregnancy: A retrospective multi-institutional study in Japan

AIMS To determine differences in pregnancy outcomes including diabetic complications, maternal and perinatal complications between gestational diabetes mellitus and overt diabetes in pregnancy in Japan. METHODS A multi-institutional retrospective study compared pregnancy outcomes between gestational diabetes mellitus and overt diabetes in pregnancy. We examined pregnant women who met the former criteria for gestational diabetes mellitus and received dietary intervention with self-monitoring of blood glucose with or without insulin. Overt diabetes in pregnancy was defined as ≥2 abnormal values on 75-g oral glucose tolerance test, fasting glucose ≥126 mg/dl (7.0 mmol/l) and 2-h postprandial glucose ≥200 mg/dl (11.1 mmol/l), or glycated hemoglobin levels ≥6.5% (48 mmol/mol). RESULTS Data were collected on 1267 women with gestational diabetes and 348 with overt diabetes in pregnancy. Pregestational body mass index was higher (26.2 ± 6.1 vs. 24.9 ± 5.7 kg, P<0.05) and gestational age at delivery was earlier (37.8 ± 2.5 weeks vs. 38.1 ± 2.1 weeks, P<0.05) in overt diabetes than in gestational diabetes. Glycated hemoglobin (6.8 ± 1.1% [51 mmol/mol] vs. 5.8 ± 0.5% [40 mmol/mol], P<0.05) and glucose on 75-g oral glucose tolerance test and prevalence of retinopathy (1.2% vs. 0%, P<0.05) and pregnancy-induced hypertension (10.1% vs. 6.1%, P<0.05) were higher in overt diabetes than in gestational diabetes. Pregnancy-induced hypertension was associated with pregestational body mass index, gestational weight gain, chronic hypertension, and nulliparity but not with 75-g oral glucose tolerance test. CONCLUSIONS Overt diabetes in pregnancy is significantly associated with maternal complications such as retinopathy and pregnancy-induced hypertension.

Proliferation and differentiation of pancreatic β-cells: ultrastructural analysis of the pancreas in diabetic mice induced by selective alloxan perfusion

To clarify the mechanism of regenerative processes of pancreatic β-cells, we constructed a new diabetic model of mice and investigated their pancreatic endocrine cells by electron microscopy. Male ICR mice (8 weeks old) were partially and chemically depancreatized by perfusing alloxan (100 mg/kg body weight) via the caudal vein after clamping the cranial mesenteric artery. By this method, we could render the mice diabetic by partial reduction of β-cells localized in the splenic, gastric, and parabiliary segment. Glucose intolerance gradually ameliorated without any treatment. In the perfused segments, pancreatic β-cells showed pyknosis and the mitochondria were swollen 6h after the treatment, while non-β-cells including α-cells remained intact. At 5 days, β-cells were few and the islets became smaller in size. At 20 weeks, small islet cell clusters (ICCs) were observed budding from interlobular and intralobular ductal epithelial cells. β-cells scattering in the exocrine pancreas were also frequently observed. In the alloxan-nonperfused segment, β-cells with thin rough endoplasmic reticulum and immature secretory granules without an electron-opaque halo were observed, and the number of mitochondria increased in some β-cells at 1 day and 5 days after the treatment. At 20 weeks, β-cells that contained only mature granules were observed in hypertrophic islets. In this model, both proliferation of residual β-cells and differentiation of pancreatic endocrine cells from the ductal epithelial cells were recognized.

Present status of clinical care for postpartum patients with hypertensive disorders of pregnancy in Japan: findings from a nationwide questionnaire survey

Objective: To assess the present status of clinical care for postpartum patients with hypertensive disorders of pregnancy (HDP) in Japan. Methods: We conducted a nationwide questionnaire survey of obstetricians, internists and hypertension specialists and analyzed 686 valid responses. Results: Though HDP is widely known as a risk factor for subsequent hypertension and cardiovascular disease, over one-third of obstetricians terminated their postpartum follow-up of HDP patients without referring them to other departments. Conclusion: It is important to establish an effective referral system, whereby patients with HDP can be smoothly transferred to primary care or a specialist physician after childbirth for long-term monitoring and management of blood pressure.

Mitochondrial DNA mutations in pancreatic biopsy specimens from IDDM patients

We investigated the contribution of mitochondrial DNA mutations to the pathogenesis of IDDM by analyzing mitochondrial DNA in pancreatic biopsy specimens and peripheral blood cells from 18 patients with newly-diagnosed IDDM. All patients presented with typical abrupt onset of diabetes and ketosis on initial examination. Point mutations at nucleotides 3243, 3271 and 8344 were assayed by polymerase chain reaction and restriction fragment length polymorphism analysis or by mismatch-primer analysis. A common large deletion from nucleotides 8483-13459 was analyzed by a primer shift method. All of these mutations are known to be pathogenic mutations. However, none of the mitochondrial DNA mutations were detected in any of 18 IDDM patients. Several types of mitochondrial DNA mutation have been identified in the peripheral blood cells in some patients with non-insulin-dependent diabetes mellitus as well as in some with IDDM, however, our results suggest that abrupt-onset IDDM does not correlate with any of the known mitochondrial DNA mutations.

Emerging therapeutic strategies in autoimmune diabetes: aetiology, prediction, prevention and cure

(1999). Emerging therapeutic strategies in autoimmune diabetes: aetiology, prediction, prevention and cure. Emerging Therapeutic Targets: Vol. 3, No. 1, pp. 177-193.