bluuebunny/crossref_metadata_2025 huggingface dataset:
In Vivo Expression of B:9‐23 Peptide/I‐Ag7 Complex May Abrogate the Inhibition of Diabetes Induced by RGD‐Fiber‐Mutant Adenovirus in NOD Mice
Authors: ABIRU NORIO , SUN FUYAN , KAWASAKI EIJI , YAMASAKI HIRONORI , OSHIMA KATSUYA , NAGAYAMA YUJI , MIZUGUCHI HIROYUKI , HAYAKAWA TAKAO , MIAO DONGMEI , LIU EDWIN , EISENBARTH GEORGE S. , EGUCHI KATSUMI
Published: January 2004

Abstract: Insulin B chain peptide B:9‐23 given to NOD mice decreases the development of diabetes, and phase II trials of an altered peptide ligand of B:9‐23 are under way in humans. We have created a gene for the NOD MHC class II β chain, covalently linked to the B:9‐23 peptide. B lymphoma cells transfected with the gene stimulated NOD islet‐derived B:9‐23 reactive T cell clones in vitro. In this study, we generated an RGD‐fiber‐mutant adenovirus vector encoding the covalent B:9‐23 peptide/I‐Ag7 gene (Ad‐RGD‐B:9‐23) to test whether in vivo expression of the gene could protect NOD mice from diabetes. NOD female mice were injected intramuscularly with 5 × 108 PFU of Ad‐RGD‐B:9‐23 and empty RGD‐adenovirus vector. A single administration of the empty vector did not alter the expression of insulin autoantibodies, but delayed the onset of diabetes in NOD mice. In contrast, Ad‐RGD‐B:9‐23 immunization induced an early expression of insulin autoantibodies, but did not change the disease occurrence compared to control NOD mice. Our results suggest that adenovirus infection could confer protection from diabetes in NOD mice. The in vivo expression of covalent B:9‐23 peptide/class II complex by adenovirus gene transfer might activate anti‐insulin autoimmunity, resulting in abrogation of the inhibition of diabetes induced by an RGD‐fiber‐mutant adenovirus vector.
Abnormal Peripheral Blood Dendritic Cell Populations in Type 1 Diabetes
Authors: PENG RUIHUA , LI Y. , BREZNER K. , LITHERLAND SALLY , CLARE‐SALZLER M. J.
Published: January 2004

Abstract: Type 1 diabetes (T1D) is a T cell‐mediated disease. Various DC populations play important roles in initiating and directing T cell responses and thus may be critical for T1D pathogenesis. We thus examined peripheral blood DC1 and DC2 populations by flow cytometry in healthy controls, subjects at risk for T1D, new‐onset patients, and established T1D patients. We found a significant increase in the number of DCs (including DC1 and DC2) in at‐risk subjects and those with new‐onset T1D versus healthy controls and established T1D patients (ANOVA; p < 0.0001). Analysis of DC1 and DC2 subsets in these same groups demonstrated a significant decrease in the ratio of DC1 and DC2 in subjects at risk and new‐onset and established T1D patients in contrast with healthy controls (p < 0.0001). Both subsets of peripheral blood DCs from T1D patients expressed significantly higher levels of HLA‐DR than healthy controls. Peripheral blood mononuclear cells from T1D patients secreted significantly higher amounts of IFN‐α than controls, and IFN‐α production correlated inversely with the DC1/DC2 ratio. This study demonstrates a marked increase in peripheral blood DC numbers that occurs during a time of active autoimmunity in at‐risk subjects and patients with new‐onset T1D, but is lost in established diabetes. However, the abnormal distribution of peripheral blood DC populations appears to be a persistent phenotype in all stages of T1D.
Characterization of Dendritic Cells in Humans with Type 1 Diabetes
Authors: SUMMERS KELLY L. , BEHME MARGARET T. , MAHON JEFFERY L. , SINGH BHAGIRATH
Published: January 2004

Abstract: The characterization of dendritic cells (DCs) in diabetes has primarily examined in vitro‐generated DCs. In this study, we have compared the composition and phenotype of naturally occurring DCs within the peripheral blood of subjects with type 1 diabetes, latent‐onset autoimmune diabetes in adults, and nondiabetic controls. We find that circulatory DC subsets exist in normal frequencies and phenotypic states in diabetic patients. In vivo, DCs were located around the pancreatic islets in type 1 diabetic patients, but were absent in pancreatic tissue of normal controls. These findings provide new insight toward understanding the pathological role of DCs in type 1 diabetes.
NKT Cell Frequency in Japanese Type 1 Diabetes
Authors: OIKAWA YOICHI , SHIMADA AKIRA , YAMADA SATORU , MOTOHASHI YOSHIKO , NAKAGAWA YOSHINORI , IRIE JUN‐ICHIRO , MARUYAMA TARO , SARUTA TAKAO
Published: January 2004

Abstract: A numerical and functional deficit of natural killer T (NKT) cells has been reported to be associated with the pathogenesis of Caucasian patients with type 1 diabetes. However, a conflicting finding of a higher frequency of NKT cells (Valpha24+ Vbeta11+ T cells) was observed in islet‐associated Ab+ and Ab− Japanese “classic” type 1 diabetes. Here, we combined the data of NKT cell frequency in Ab+ and Ab−“classic” type 1 diabetic patients and then analyzed the relationship between NKT cell frequency and disease activity.
IL‐6 Modulates Hyperglycemia‐Induced Changes of Na+ Channel Beta‐3 Subunit Expression by Schwann Cells
Authors: SKUNDRIC DUSANKA S. , DAI RUJUAN , MATAVERDE PHILIP
Published: January 2004

Abstract: Our data show the suppressive effect of hyperglycemia on expression of voltage‐gated Na+ channel β3 subunit by Schwann cells as well as the protection of β3 expression by treatment with IL‐6. Our findings have direct relevance for therapy of diabetic neuropathy.
Glucose‐Responsive Expression of the Human Insulin Promoter in HepG2 Human Hepatoma Cells
Authors: BURKHARDT BRANT R. , LOILER SCOTT A. , ANDERSON JO ANNE , KILBERG MICHAEL S. , CRAWFORD JAMES M. , FLOTTE TERENCE R. , GOUDY KEVIN S. , ELLIS TAMIR M. , ATKINSON MARK
Published: January 2004

Abstract: The concept of insulin production afforded by hepatic gene therapy retains promise as a potential therapy for type 1 diabetes, but the approach has been limited by the need for strict transgene regulation in response to fluctuating levels of both glucose and insulin. Furthermore, while hepatocytes contain various glucose‐responsive elements, they lack the appropriate regulated secretory system necessary for insulin release, thereby necessitating the requirement for transcriptional regulation of hepatic insulin production under the direction of a glucose‐responsive promoter. To address this, we have evaluated several glucose‐responsive promoters that may be used successfully for hepatic insulin production via recombinant adeno‐associated virus (rAAV) therapy. Our results suggest that the human insulin promoter represents a strong candidate as a robust, glucose‐responsive promoter for regulated hepatic insulin production.
Interleukin‐6 Protects MIN6 β Cells from Cytokine‐Induced Apoptosis
Authors: PARK HYEWON , AHN YUHERN , PARK CHUNG‐KYU , CHUNG HEE‐YONG , PARK YONGSOO
Published: January 2004

Abstract: Host nonspecific β cell injury by cytokines has been implicated in the process of early islet graft dysfunction. Islet transplant destruction by cytokines released from inflammatory cells that infiltrate the graft is thought to be mainly mediated by NF‐κB‐dependent nitric oxide (NO) production by the islet. In this study, we aimed to evaluate the role of IL‐6 in making a β cell resistant to cytokine‐induced apoptosis by decreasing the NF‐κB‐dependent NO production and compared the result with that of the expression of a dominant negative inhibitor of NF‐κB (DN NF‐κB). Incubation of MIN6 cells with IL‐1β, IFN‐γ, and TNF‐α (cytomix) increased production of nitrites, with increased expression of iNOS mRNA. When treated with cytomix, the DN NF‐κB‐transfected mutant demonstrated significantly less nitrite production and apoptosis than parent MIN6. NO production was effectively blocked by IL‐6 as well as by N‐monomethyl‐l‐arginine (l‐NMMA). Inhibition of the NO production led to decreased rate of apoptosis accompanied by downregulation of the proapoptotic molecule Bax and increased expression of the antiapoptotic molecule Bcl‐2 and Bcl‐xL. These data indicate that cytokine‐induced cell death in the MIN6 β cell line involves mechanisms that are, in part, NF‐κB‐ and NO‐dependent. Inhibition of the NO production by the incubation of the MIN6 cells by the pretreatment of IL‐6 or l‐NMMA is cytoprotective and can be used as a substitute for the expression of DN NF‐κB.
A Diabetes‐Related Epitope of GAD65
Authors: MYERS MARK A. , FENALTI GUSTAVO , GRAY ROBYN , SCEALY MARITA , TONG JONATHAN C. , EL‐KABBANI OSSAMA , ROWLEY MERRILL J.
Published: January 2004

Abstract: The 65‐kDa isoform of glutamic acid decarboxylase (GAD65) is a major autoantigen in type 1 diabetes, and most patients have serum antibodies reactive with conformational epitopes on the GAD65 molecule. The aims of this study were to prepare mutants of GAD65 to further localize the type 1 diabetes epitope in the region of the PEVKEK loop of GAD65 and to identify the particular amino acids within the epitope that are recognized by autoimmune diabetes sera.
SOX13 Autoantibodies Are Likely to Be a Supplementary Marker for Type 1 Diabetes in Korea
Authors: PARK YONGSOO , PARK HYEWON , YOO EUNKYUNG , KIM DUKHEE
Published: January 2004

Abstract: The SOX13, one of the family of transcription factors that play key roles in organ development, is reported to be a diabetes autoantigen, islet cell antigen 12 (ICA12). Recently, a study of antibodies to SOX13 was conducted in patients with type 1 diabetes mellitus (T1DM) indicating that these antibodies potentially identified patients without antibodies to the major T1DM‐associated autoantigens, insulin, GAD, or IA‐2. We know that the prevalence of islet‐specific autoantibodies (GAD, IA‐2) in Korean patients is much lower than that in white patients. It may be possible that other autoantibodies that could be directed to as yet unknown antigen may play a role in Korean T1DM patients. To investigate this, we measured SOX13 autoantibodies applying a radioligand binding assay using in vitro transcribed and translated antigen in 188 T1DM patients (mean duration, 4.2 years) and 64 T2DM patients and compared the results with those of 101 healthy control subjects. SOX13 autoantibodies occurred at a significantly higher frequency among T1DM patients (55/188, 29.3%) than among T2DM patients (4/64, 6.2%) or healthy adult controls (1/101, 1%). The 55 patients with positive SOX13 antibodies had significantly shorter duration of diabetes than SOX13 antibody‐negative patients (3.6 ± 2.8 vs. 4.5 ± 3.9 years; p < 0.05). We could detect a prevalence similar to control in patients with Hashimoto's thyroiditis (4.9%, n= 101) and rheumatoid arthritis (6.7%, n= 89). As a whole, 44 of the 55 patients with SOX13 antibodies had at least one or more other autoantibodies to the major T1DM‐associated autoantigens. However, SOX13 antibodies were the only antibodies detected as positive in 1 of the 11 new‐onset patients. We conclude, therefore, that these antibodies are likely to be one of several epitope‐spreading responses to islet‐ or nonislet‐specific autoantigens seen in the development of T1DM, and they may be used as a supplementary marker for investigating T1DM in Korea.
Development and Validation of a Radioligand Binding Assay to Measure Insulin Specific IgG Subclass Antibodies in Human Serum
Authors: FINCO‐KENT D. , MORRONE A. , MOXNESS M. , BEDIAN V. , KRASNER A. , FOLEY J. , STENE M. , KAWABATA T.
Published: January 2004

Abstract: The objective was to develop and validate a radioligand binding assay for insulin antibodies (IABs) of the IgG1, IgG2, IgG3, and IgG4 subclasses in human serum. The validation studies focused on determining specificity, capacity, linearity, sensitivity, and precision of each assay. It was seen that our assay for IAB IgG subclasses is specific and has sufficient capacity to measure each of the subclasses in human serum. Moreover, the linear region and limits of detection and quantitation for each assay are clearly determined.
Development and Validation of Radioligand Binding Assays to Measure Total, IgA, IgE, IgG, and IgM Insulin Antibodies in Human Serum
Authors: MOXNESS MICHAEL , FOLEY JIM , STENE MARK , FINCO‐KENT DEBORAH , BEDIAN VAHE , KRASNER ALAN , KAWABATA THOMAS
Published: January 2004

Abstract: Radioligand binding assays for total and Ig classes of insulin antibodies (IAB) were developed and validated. For each assay, insulin‐extracted serum samples were incubated with radiolabeled insulin in the presence and absence of high levels of unlabeled insulin to determine nonspecific binding and total binding, respectively. To measure total IAB, antibody‐bound insulin was precipitated with a polyethylene glycol solution, washed, and counted in a gamma‐counter. To measure IgG IAB, samples were treated with protein G‐Sepharose beads, centrifuged, washed, and counted. For the measurement of IgA, IgE, and IgM IAB, IgG was removed from the samples and treated with anti‐IgA, ‐IgE, or ‐IgM conjugated to Sepharose beads, centrifuged, washed, and counted. The acid/charcoal extraction of bound and unbound insulin from serum samples was optimized. Specificity and binding capacity of the protein G and antibody‐bound beads were evaluated and optimized. The linear region of the total and IgG IAB assays was determined using serum samples containing high levels of insulin antibodies. The limit of quantitation, limit of detection, and precision for all the assays were also determined.
Induction of Diabetes‐Related Autoantibodies below Cutoff for “Positivity” in Young Nondiabetic Children
Authors: HOLMBERG HANNA , VAARALA OUTI , FÄLTH‐MAGNUSSON KARIN , LUDVIGSSON JOHNNY
Published: January 2004

Abstract: The aim was to study the natural course of diabetes‐related autoantibodies at low concentrations, below “positivity”, in a nondiabetic population followed up from infancy. Blood samples were taken from 205 children at 6 weeks, 6 months, 18 months, and 5 years of age. Autoantibodies against GAD65 (GADA), tyrosine phosphatase (IA‐2A), and insulin (IAA) were determined by radioligand‐binding assays. All children had detectable levels of GADA and approximately half had IA‐2A, but only approximately 10% had detectable levels of IAA during the follow‐up period. Many children developed IA‐2A already at 6 months of age, similar concentrations were seen at 18 months, and then the levels of IA‐2A decreased until 5 years of age. GADA were induced less often at 6 months of age, increased up to 18 months, and fluctuated at similar levels up to 5 years of age. IAA were detectable in so few children and at low levels, so no trend in natural course could be revealed. We conclude that there is a natural induction of humoral immune response to β cell autoantigens early in life. Our results suggest that the mechanisms of β cell tolerance to GAD and IA‐2 differ in healthy children.
Cow‐Milk‐Free Diet during Last Trimester of Pregnancy Does Not Influence Diabetes‐Related Autoantibodies in Nondiabetic Children
Authors: LUDVIGSSON JOHNNY
Published: January 2004

Abstract: The aim was to study whether cow‐milk‐free diet during the last trimester of pregnancy influences the development of diabetes‐related autoantibodies in nondiabetic children. We also examined the effect of later introduction of cow milk proteins and gluten. Blood samples were taken from 205 children at 6 weeks, 6 months, 18 months, and 5 years, whose mothers had been randomized to either a cow‐milk‐free diet or not during the last trimester of pregnancy. During the first 3 months of life, cow milk proteins were not fed to the children. Autoantibodies against GAD.65 (GADA), tyrosine phosphatase (IA‐2A), and insulin (IAA) were determined by a radioligand‐binding assay. We found specific autoantibodies, although in low concentrations below the traditional cutoff for positivity in the children. The diet of the mother during pregnancy had no influence on development of autoantibodies, nor did duration of breast‐feeding or time for introduction of gluten. Cow milk introduction between 3 and 6 months caused a transient increase in GADA at 6 months of age (p < 0.05). Children who developed atopic disease had significantly lower IA‐2A at 6 months (p < 0.001).
Difference in Gene Expression Profiles between Human CD4+CD25+ and CD4+CD25− T Cells
Authors: PATI NIRUPMA , GHOSH SOUMITRA , HESSNER MARTIN J. , KHOO HUOY‐JII , WANG XUJING
Published: January 2004

Abstract: We studied the gene expression profiles of human CD4+CD25+ and CD4+CD25− T cells by using cDNA microarrays. Our preliminary results indicate that there are likely significant differences in the regulation of apoptosis, cell cycle, cytokine receptor, cell‐cell interaction, and stress pathway genes between these two subtypes of T cells.
The Design of a Gene Chip for Functional Immunological Studies on a High‐Quality Control Platform
Authors: WAUKAU JILL , JAILWALA PARTHAV , WANG YOUMIN , KHOO HUOY‐JII , GHOSH SOUMITRA , WANG XUJING , HESSNER MARTIN J.
Published: January 2004

Abstract: We have created an immunology‐related microarray chip containing primarily known genes with well‐studied functional properties. By looking at known genes rather than expressed sequence tags, we hope to gain a better understanding of immunological pathways and how they work. The immunology gene chip contains genes from the following functional categories: T cell genes; B cell genes; dendritic cell genes; chemokine and cytokine genes; apoptosis genes; cell cycle genes; cell interaction genes; general hematology and immunology genes; and adhesion genes. We have also developed a novel three‐color cDNA array platform in which arrays are directly visualized before hybridization, which allows us to select only high‐quality chips for our experiments. In an effort to provide quantitative quality control for each array element as well as the entire chip, we have developed Matarray, a software package for image processing and data acquisition. With Matarray, we have built a quantitative data filtering and normalization scheme that has proved to be more efficient than the existing methods. The list of immunology chip genes is available from the authors.
Relationship between T and B Cell Responses to Proinsulin in Human Type 1 Diabetes
Authors: DURINOVIC‐BELLÓ IVANA , MAISEL NICOLA , SCHLOSSER MICHAEL , KALBACHER HUBERT , DEEG MARTIN , EIERMANN THOMAS , KARGES WOLFRAM , BOEHM BERNHARD O.
Published: January 2004

Abstract: In type 1 diabetes, humoral and cell‐mediated responses to insulin and proinsulin are detectable. Autoantibodies to insulin are associated with impending disease in young individuals and are used as predictive markers to determine disease risk. The aim of this study was to investigate whether different cytokine patterns of cellular reactivity to insulin might serve as additional specific markers of disease maturation and might improve disease prediction in individuals at risk. We correlated T and B cell responses to insulin in subjects with increased genetic risk (HLA‐DRB1*04, DQB1*0302) for diabetes with or without islet autoantibodies (Ab+ subjects and controls, respectively) and HLA‐matched patients. Peripheral blood mononuclear cells were stimulated with 15 overlapping proinsulin peptides (16‐mer), and proinflammatory Th1 (IFNγ) and anti‐inflammatory Th2 (IL‐4) cytokines were analyzed. We observed a simultaneous increase in IL‐4 and IFNγ secretion in early islet autoimmunity of Ab+ subjects, but not in insulin‐treated T1D patients. Furthermore, the increase in IL‐4 secretion in Ab+ subjects was associated with insulin autoantibody responses. There was no correlation of either IFNγ or IL‐4 secretion with insulin antibody responses in patients already treated with exogenous insulin. In conclusion, our findings reveal that quantification of cytokine responses to proinsulin in peripheral blood may prove to be a promising specific marker of diabetes progression and could, in addition to insulin autoantibodies, be used in the prediction of type 1 diabetes.
Inheritance of MHC Class II Genes in Lithuanian Families with Type 1 Diabetes
Authors: SADAUSKAITE‐KUEHNE VAIVA , VEYS KEN , LUDVIGSSON JOHNNY , PADAIGA ZILVINAS , SANJEEVI CARANI B.
Published: January 2004

Abstract: Type 1 diabetes mellitus (DM) is caused by genetic and environmental factors. Twice as many fathers as mothers of children with type 1 DM have the disease. The reason for the differences remains unclear. We looked at the transmission rates of diabetes‐related alleles from parents to children with diabetes. All children with newly diagnosed type 1 DM from August 1, 1996 to August 1, 2000, aged 0 to 15 years, in Lithuania were invited to participate. Blood samples for full genetic analysis were available from 125 families. HLA DQA1, DQB1, and DRB1 typing was done on DNA extracted from peripheral blood, by polymerase chain reaction amplification, manual dot‐blotting onto nylon membranes, synthetic sequence‐specific oligonucleotide probe 3′‐end labeling with 32P‐dCTP, and hybridization, followed by stringency washes, autoradiography, and allele calling. Frequency of diabetes risk‐related alleles DQB1*0302, DQA1*0201, DR4, and DR3 was less prevalent among Lithuanian than among Swedish children with type 1 DM. Transmission rates of DR4‐DQB1*0302‐DQA1*0301 and DR3‐DQB1*0201‐DQA1*0501 haplotypes from parents were higher than expected: χ2 (TDT) 30.56, p < 0.0001, and χ2 (TDT) 11.26, p= 0.0008, respectively. DQB1*0302 and DR4 were significantly more frequently transmitted from both parents, but DR3 was transmitted more frequently only from mothers. Any of these alleles had similar frequencies among female and male offspring. We conclude that, besides DR4‐DQB1*0302‐DQA1*0301 and DR3‐DQB1*0201‐DQA1*0501, there are other inherited alleles that determine risk for type 1 DM among children in Lithuania. Fathers might transfer other alleles of disease susceptibility in higher frequency or mothers might provide a protective environment during pregnancy, which results in higher risk to offspring of fathers than mothers to develop diabetes.
Ethnic Differences in the Associations between the HLA‐DRB1*04 Subtypes and Type 1 Diabetes
Authors: REWERS AMANDA , BABU SUNANDA , WANG TIAN BAO , BUGAWAN TEODORICA L. , BARRIGA KATHY , EISENBARTH GEORGE S. , ERLICH HENRY A.
Published: January 2004

Abstract: The HLA genotype DRB1*03,DQB1*0201/DRB1*04,DQB1*0302 confers a 25‐fold increase in the risk of type 1 diabetes. In persons with this genotype, DRB1*0405, *0402, and *0401 subtypes have been reported to further increase risk, whereas the *0403 and *0406 alleles confer a relative protection. We compared the frequencies of the DRB1*04 alleles in 193 type 1 diabetic patients with the HLA‐DRB1*03,DQB1*0201/DRB1*04,DQB1*0302 genotype (140 non‐Hispanic white [NHW] and 53 Hispanic) and 205 nondiabetic controls (142 NHW and 63 Hispanic). In addition, 87 NHW first‐degree relatives of type 1 diabetes patients were studied: 33 positive and 54 negative for autoantibodies to insulin, GAD65, or IA‐2. The HLA‐DRB1 was typed using standard PCR SSOP methods. DRB1*0401 (OR, 2.19; 95% CI, 1.36‐3.54) in NHW and *0405 (OR, 3.78; 95% CI, 1.43‐10.0) in Hispanics were significantly associated with T1DM, whereas DRB1*0403 was protective (OR, 0.19; 95% CI, 0.04‐0.89 in NHWs; OR, 0.10; 95% CI, 0.01‐0.83 in Hispanics). Associations between the DRB1*04 alleles and prediabetic islet autoimmunity were generally in the same direction as those with diabetes. Among diabetic patients, the mean age of diagnosis appeared to be higher among those with the *0403 and *0407 allele compared with the others. In summary, on the DRB1*03,DQB1*0201/DRB1*04,DQB1*0302 genotypes, the *0403 allele confers relative protection from type 1 diabetes and development of islet autoantibodies in both Hispanics and NHWs and is associated with older age at diabetes diagnosis. Although the associations between diabetes and *0401 and *0405 appear to differ somewhat between Hispanics and NHWs, overall there is no significant difference between these two ethnic groups.
MIC‐A Genotypes 4/5.1 and 9/9 Are Positively Associated with Type 1 Diabetes Mellitus in Brazilian Population
Authors: TICA VALERIA , NIKITINA‐ZAKE LIENE , DONADI EDUARDO , SANJEEVI CARANI B.
Published: January 2004

Abstract: The aim of our study was to evaluate the frequencies of MIC‐A alleles and genotypes in Brazilian patients with type 1 diabetes mellitus (T1DM) and healthy controls. MHC class I chain‐related gene‐A (MIC‐A) has been shown to be associated with susceptibility to T1DM in different populations. We analyzed the DNA samples from 86 patients and 201 healthy controls for MIC‐A by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer. We found increased frequencies of two genotypes, MIC‐A 4/5.1 (pc= 0.006; OR, 6.1) and 9/9 (pc= 0.045; OR, 5.75), in our patients (mean diagnosis age, 11.70 years; SD, 8.86; mean age, 20.44 years; SD, 12.17) compared with the controls (median age, 26.41 years; SD, 8.87).
HLA‐DRB1 and MICA in Autoimmunity
Authors: BILBAO J. RAMÓN , MARTÍN‐PAGOLA AINHOA , PÉREZ DE NANCLARES GUIOMAR , CALVO BEGOÑA , VITORIA JUAN CARLOS , VÁZQUEZ FEDERICO , CASTAÑO LUIS
Published: January 2004

Abstract: Autoimmune disorders such as type 1 diabetes (T1DM), celiac disease (CD), and Addison's disease (ADD) develop in individuals with genetic susceptibility that are exposed to environmental triggering factors not completely defined. Patients with an autoimmune disease (and their relatives) are at increased risk of developing another disorder, and this might be caused by a common genetic origin of autoimmunity; for example, HLA class II region in 6p21 shows a very strong association with most diseases. The aim of this study was to determine whether shared susceptibility markers extend from the central (DRB1) through the telomeric (MICA) HLA region. We analyzed three independent sets of families with one autoimmune disease, T1DM, CD, or ADD, and genotyped them for HLA‐DRB1 and for the exon 5 GCT polymorphism of MICA. For HLA‐DRB1, allele DRB1*0301 was the only one associated with risk for all three diseases; in the case of MICA, allele A9 was found to be the common protective allele. Haplotype analysis shows that haplotype A5.1‐DRB1*0301 confers risk to autoimmunity. Our results show that there are common risk and protection alleles in both loci, suggesting a core of genetic association with autoimmunity (HLA‐DRB1*0301 risk; A9 protection) that could be modulated by other alleles/loci or environmental factors toward one or another disease. Some alleles are part of conserved haplotypes (A5.1‐DR3, A5.1‐DR2), whereas others seem to have independent effect (A9) and support the idea of two independent loci in this region.
5′‐Insulin Gene VNTR Polymorphism Is Specific for Type 1 Diabetes
Authors: PÉREZ DE NANCLARES GUIOMAR , BILBAO J. RAMÓN , CALVO BEGOÑA , VITORIA JUAN CARLOS , VÁZQUEZ FEDERICO , CASTAÑO LUIS
Published: January 2004

Abstract: The VNTR region located at the 5′‐end of the insulin gene on chromosome 11p15.5 is linked to susceptibility to type 1 diabetes mellitus (T1DM), and class I alleles have been associated with increased risk of disease, whereas class III alleles are considered to be protective. Although a potential effect on the expression level of thymic insulin and a consequent abnormal tolerance have been proposed as an explanation, it is still not clear whether the association is specific for T1DM or whether it is shared by other autoimmune disorders. To investigate the contribution of INS‐VNTR to the genetic susceptibility to autoimmune disorders, we analyzed 102 autoantibody‐positive T1DM patients, 59 patients with celiac disease (CD), and 57 patients with Addison's disease (ADD), as well as 111 unrelated healthy individuals from the general population. When analyzing the results, class I allele frequencies were 85.8% in the T1DM group, 77% among CD patients, 71% in the ADD group, and 76.1% in the general population. Association with increased risk was seen only in the T1DM group (pc= 0.015). Risk to T1DM was associated with the class I/class I homozygous genotype (RR, 1.92; 95% CI, 1.03‐3.6). In conclusion, INS‐VNTR does not seem to be involved in the susceptibility to autoimmune diseases other than T1DM and can be considered a diabetes‐specific locus.
Nicotinamide Adenine Dinucleotide Phosphate Oxidase (NADPH Oxidase) P22 Phox C242T Gene Polymorphism in Type 1 Diabetes
Authors: MATSUNAGA SEIKO , MARUYAMA TARO , YAMADA SATORU , MOTOHASHI YOSHIKO , SHIGIHARA TOSHIKATSU , SHIMADA AKIRA , SARUTA TAKAO
Published: January 2004

Abstract: Type 1 diabetes is caused by the immune‐mediated destruction of insulin‐secreting pancreatic β cells and is thought to be an autoimmune disease resulting from a complex interaction of genetic and environmental factors. In animal models of type 1 diabetes, macrophages and their products, superoxides, have central roles in the β cell destruction, but in humans their roles remain unclear. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase produces superoxide in macrophages, and its essential component, p22 phox, is a critical enzyme for superoxide production. The C242T polymorphism in the p22 phox coding gene has been reported to be associated with reduced oxidase activity. We therefore investigated whether the p22 phox gene polymorphism affected the susceptibility to and clinical course of type 1 diabetes. We examined 287 Japanese type 1 diabetic patients and 425 unrelated nondiabetic subjects. In addition, we allocated the diabetic patients to the following three groups: (1) acute‐onset type 1 diabetes with at least one autoantibody (GADA, IA‐2, IAA); (2) acute‐onset type 1 diabetes without autoantibodies; and (3) slow‐onset type 1 diabetes with autoantibody. We could not find a significant difference in p22 phox genotype and T allele frequency between overall type 1 diabetic patients and control subjects. Regardless of the onset pattern and autoantibody positivity of type 1 diabetes, no difference in p22 phox genotype and T allele frequency was found among the groups. In conclusion, the p22 phox C242T gene polymorphism did not affect the susceptibility to and clinical course of Japanese type 1 diabetes.
Stromal Cell‐Derived Factor‐1 Chemokine Gene Polymorphism Is Not Associated with Onset Age of Japanese Type 1 Diabetes
Authors: SHIGIHARA TOSHIKATSU , SHIMADA AKIRA , YAMADA SATORU , MARUYAMA TARO , HIROSE HIROSHI , SARUTA TAKAO
Published: January 2004

Abstract: Type 1 diabetes is characterized by cell‐mediated autoimmune destruction of pancreatic beta cells. Although the disease shows a strong association with HLA class II alleles, other genes may influence the initiation or the rate of progression of the autoimmune process. Recently, it was reported that a polymorphism of the stromal cell‐derived factor‐1 (SDF‐1) (a kind of chemokine) gene was associated with early onset of type 1 diabetes in Caucasians. Therefore, we examined SDF‐1 gene polymorphism in Japanese type 1 diabetes in this study. We examined the SDF‐1 gene polymorphism (801G→A) in 298 unrelated Japanese type 1 diabetic patients and 270 healthy subjects by the TaqMan PCR method. Allelic and genotypic frequencies of the SDF‐1 A variants were similar in overall type 1 diabetic patients and healthy subjects. We then stratified the patients by their onset pattern (acute vs. slow onset) and islet‐associated autoantibody positivity. However, no significant difference was found among each group of type 1 diabetes. Furthermore, unlike the previous report in “Caucasian” type 1 diabetics, the SDF‐1 A variant was not associated with early onset of the disease in Japanese type 1 diabetics. The SDF‐1 gene polymorphism was not associated with onset age (or onset pattern) of type 1 diabetes in Japanese. Further study is necessary to conclude whether SDF‐1 gene polymorphism affects the onset age in type 1 diabetes in general.
Mutation Scan of a Type 1 Diabetes Candidate Gene
Authors: NOLSØE RÚNA L. , POCIOT FLEMMING , NOVICK DANIELA , RUBINSTEIN MENACHEM , KIM SOO‐HYUN , DINARELLO CHARLES A. , MANDRUP‐POULSEN THOMAS
Published: January 2004

Abstract: Type 1 (insulin‐dependent) diabetes, T1DM, is the result of an immune‐mediated destruction of the pancreatic β cells dependent mainly on T helper cells and macrophages. Interleukin‐18 (IL‐18) is a proinflammatory cytokine produced mainly by macrophages. IL‐18 is capable of inducing T lymphocyte synthesis of IFNγ, thereby skewing the T helper response toward a T helper type 1 (Th1) profile. IL‐18 binding protein (IL18BP) neutralizes IL‐18 and leads to a reduced Th1 response. Polymorphisms in IL18BP may affect the activity of IL‐18 and the magnitude of the Th1 response and may play a role in the pathogenesis of T1DM. The aim of the study was therefore to identify polymorphisms in IL18BP and to test these for association with T1DM. We evaluated the human IL18BP gene on chromosome 11q13 as a candidate susceptibility gene for T1DM and scanned the entire IL18BP (promoter, exons 1‐6, and 3′UTR) for polymorphisms using single‐strand conformational polymorphism analysis and direct sequencing. We identified a total of 11 polymorphisms, all having allele frequencies ranging between 0.05 and 0.10. Four were in the 5′UTR: −257G→T, −78C→T, −65G→A, and −59A→G. Three were in intron 3: IVS3+140A→C, IVS4−147G→T, and IVS4−59G→T. The last four, 38*A→T, 48*T→A, 388*C→G, and 440*_441*insG, were in the 3′UTR of IL18BP. However, none of these were frequent enough to permit association studies in T1DM and we conclude that IL18BP does not contribute to the overall genetic susceptibility to type 1 diabetes.
Caspase 7 Is a Positional Candidate Gene for IDDM 17 in a Bedouin Arab Family
Authors: BABU SUNANDA R. , BAO FEI , ROBERTS CHRISTINE M. , MARTIN ALLISON K. , GOWAN KATHERINE , EISENBARTH GEORGE S. , FAIN PAMELA R.
Published: January 2004

Abstract: The IDDM 17 locus was mapped to an 8‐cM interval at chromosome 10q25.1 based on linkage in a large Bedouin Arab family with 19 affected relatives. Caspase 7 (CASP7), an apoptosis‐related cysteine protease, is one of the few known genes in this region. CASP7 is involved in the activation cascade of caspases responsible for apoptosis execution. Only 1 of the 18 SNPs in CASP7 (SNP144692) differed significantly in frequency in the haplotypes found in affected individuals compared to control Bedouin haplotypes. This same SNP showed evidence of association with diabetes in a subset of patients (DR3/DR4*0302) from HBDI families.
Interleukin‐10 Gene Promoter Region Polymorphisms in Patients with Type 1 Diabetes and Autoimmune Thyroid Disease
Authors: IDE AKANE , KAWASAKI EIJI , ABIRU NORIO , SUN FUYAN , FUKUSHIMA TETSUYA , ISHII REIKO , TAKAHASHI RYOKO , KUWAHARA HIRONAGA , FUJITA NARUHIRO , KITA ATSUSHI , IMAIZUMI MISA , OSHIMA KATSUYA , USA TOSHIRO , UOTANI SHIGEO , EJIMA ERI , YAMASAKI HIRONORI , ASHIZAWA KIYOTO , YAMAGUCHI YOSHIHIKO , EGUCHI KATSUMI
Published: January 2004

Abstract: Type 1 diabetes is a heterogeneous autoimmune disease and is frequently associated with other organ‐specific autoimmune diseases, including autoimmune thyroid disease (AITD). Type 1 diabetic patients with AITD are known to show distinct clinical and immunological features from patients without AITD. This study investigated whether interleukin‐10 (IL‐10) gene promoter region polymorphisms are associated with susceptibility to type 1 diabetes and AITD. The frequency of −1082G/A, −819C/T, and −592C/A polymorphisms was analyzed in 54 type 1 diabetic patients with AITD, 74 type 1 diabetic patients without AITD, 124 nondiabetic patients with AITD, and 107 healthy subjects in a case‐control study. No significant differences on the allele and genotype frequencies of three polymorphisms were found not only in type 1 diabetic patients with AITD compared with normal controls, but also between nondiabetic patients with AITD and healthy controls. The distribution of IL‐10 gene haplotypes was also similar between both patient groups and normal controls. These results suggest that IL‐10 gene promoter region polymorphisms are not associated with genetic susceptibility to type 1 diabetes and AITD.
Single Nucleotide Polymorphism Study of IDDM 17 in a Bedouin Arab Family
Authors: BAO FEI , BABU SUNANDA R. , ROBERTS CHRISTINE M. , MARTIN ALLISON K. , GOWAN KATHERINE , EISENBARTH GEORGE S. , FAIN PAMELA R.
Published: January 2004

Abstract: Type 1 diabetes is an autoimmune disease caused by a combination of genetic and environmental factors. On the basis of a genomic search for linkage in a Bedouin Arab family with 19 members with type 1 diabetes, we previously mapped the IDDM 17 locus to the chromosome 10q25.1 region. The result from a recent genome scan showed suggestive evidence of linkage of IDDM 17 in a subset of Caucasian families in which all affected individuals have DR3, indicating that the IDDM 17 locus might have a measurable effect in Caucasian populations from the United Kingdom and the United States. High‐resolution SNP typing provides strong evidence of linkage disequilibrium to the IDDM 17 locus.
No Evidence for Linkage in Swedish Multiplex T1DM Families to IL12B on Chromosome 5q33‐34
Authors: HOLM PERNILLA , LUTHMAN HOLGER , KOCKUM INGRID
Published: January 2004

Abstract: Type 1 diabetes mellitus (T1DM) is an autoimmune disease in which the β cells in the pancreas are destroyed by the body's own immune system. IL12 plays a role in pathological situations, such as septic shock, tissue damage during inflammation, and organ‐specific autoimmune diseases. In NOD mice, administration of IL12 induces T1DM and administration of IL12 antagonists prevents T1DM. Linkage and association of IL12 to T1DM have been reported previously. We are unable to replicate this linkage on chromosome 5q33‐34 in 184 Swedish families. Further, we exclude a gene with λs > 1.4 from this region. Together with previously published findings, these data make IL12 an unlikely major susceptibility gene for T1DM.
Frequency of Latent Autoimmune Diabetes in Adults in Asian Patients Diagnosed as Type 2 Diabetes in Birmingham, United Kingdom
Authors: TICA VALERIA , HANIF M. WASIM , ANDERSSON ANNIKA , VALSAMAKIS G. , BARNETT A. H. , KUMAR SUDESH , SANJEEVI C. B.
Published: January 2004

Abstract: The aims of our study were to measure autoantibodies to glutamic acid decarboxylase and autoantibodies to protein tyrosine phosphatase in patients with type 2 diabetes mellitus, patients with impaired glucose tolerance, and healthy controls of Asian origin from Birmingham, United Kingdom. According to our findings, 27% (9/33) of patients initially diagnosed with type 2 diabetes mellitus carry autoantibodies to GAD65.
Two Cases of “Fulminant” Type 1 Diabetes Suggesting Involvement of Autoimmunity
Authors: NAKAGAWA YOSHINORI , SHIMADA AKIRA , OIKAWA YOICHI , IRIE JUN‐ICHIRO , SHIGIHARA TOSHIKATSU , TSUMURA KAZUHIRO , NARUMI SHOSAKU , SARUTA TAKAO
Published: January 2004

Abstract: Recently, a novel subtype of type 1 diabetes, so‐called fulminant type 1 diabetes, has been proposed. One of the characteristics of this subtype is the absence of detectable “islet‐associated” autoantibody, so it was originally proposed as being “nonautoimmune‐mediated”; however, it has not yet been concluded whether autoimmunity is involved. We have previously shown that serum interferon‐inducible protein‐10 and glutamic acid decarboxylase‐reactive CD4+ interferon‐γ‐producing cells in the peripheral blood are good markers for T cell‐mediated autoimmunity in type 1 diabetes. Here, we report two cases of fulminant type 1 diabetes in which these markers were detected and in which the involvement of islet‐associated autoimmunity is suggested.
Multicenter Prevention Trial of Slowly Progressive Type 1 Diabetes with Small Dose of Insulin (the Tokyo Study)
Authors: MARUYAMA TARO , SHIMADA AKIRA , KANATSUKA AZUMA , KASUGA AKIRA , TAKEI IZUMI , YOKOYAMA JUNICHI , KOBAYASHI TETSURO
Published: January 2004

Abstract: In 1996, we designed a randomized multicenter study to assess the effects of small doses of insulin on beta cell failure in slowly progressive type 1 diabetes (the Tokyo Study). We report here the preliminary results of this study. Glutamic acid decarboxylase 65 antibody (GADA)‐positive patients were randomly divided into 2 groups: one group received insulin (Ins group), the other a sulfonylurea (SU group). Fifty‐four patients (24 Ins group, 30 SU group) were analyzed at the end of a 4‐year period. All patients underwent a 75 g oral‐glucose test (O‐GTT) every 6‐12 months. The insulin‐dependent stage was defined based on an integrated value of serum C‐peptide levels on O‐GTT (∑CPR; sum of CPR at 0, 30, 60, 90, and 120 min) below 4.0 ng/mL. The ∑CPR in the SU group decreased progressively from 22.0 ± 10.6 to 11.3 ± 7.5 ng/mL over the 48‐month period (p < 0.001 vs. baseline). The ∑CPR in the Ins group was unchanged. Among the SU group, 30% of subjects (9/30) progressed to IDDM, while 8.3% of Ins group subjects (2/24) progressed to IDDM (p= 0.087). With regard to the subjects who had a preserved C‐peptide response (∑CPR ≥ 10 ng/mL), the proportion of SU group subjects who progressed to IDDM was significantly higher than that of the Ins group (7/28, 25% vs. 0/21, 0%, p= 0.015). Among subjects with a high GADA titer (≥0 U/mL), 9/14 (64.3%) of the SU group, but only 2/16 (12.5%) of the Ins group, developed IDDM (p= 0.0068). As to those with a high GADA titer and a preserved C‐peptide response, SU group subjects progressed to IDDM (7/12, 58.3%) more frequently than Ins group subjects (0/14, 0%) (p= 0.0012). In summary, our results suggest that small doses of insulin effectively prevent beta cell failure in slowly progressive type 1 diabetes. We recommend avoiding SU treatment and instead administering insulin to NIDDM patients with high GADA titer.
Late‐Onset Autoimmune Diabetes in Relatives of People with Type 1 Diabetes
Authors: FOURLANOS SPIROS , COLMAN PETER G. , HARRISON LEONARD C.
Published: January 2004

Abstract: The Melbourne Prediabetes Family Study, a prospective study of first‐degree relatives of people with type 1 diabetes (T1D), provided an opportunity to examine late‐onset autoimmune diabetes within the context of a family history of T1D. We compared genetic, immunologic, and clinical features in relatives of people with T1D, who developed early‐ versus late‐onset diabetes.
Insulin Resistance in Latent Autoimmune Diabetes of Adulthood
Authors: BEHME M. T. , DUPRÉ J. , HARRIS S. B. , HRAMIAK I. M. , MAHON J. L.
Published: January 2004

Abstract: Insulin resistance in patients with latent autoimmune diabetes of adulthood (LADA) was determined by homeostasis model assessment (HOMA). LADA was identified by a clinical phenotype of type 2 diabetes with antibodies to GAD65 and/or IA‐2/ICA512. All patients were managed with insulin therapy. Insulin resistance in LADA was lower than in antibody‐negative type 2 diabetes, higher than in normal humans and in recent‐onset type 1 diabetes, and similar to that in long‐term type 1 diabetes. Mean values for HOMA varied linearly with mean values for BMI, which accounted for much of the insulin resistance in these forms of diabetes. LADA resembles long‐term type 1 diabetes with respect to insulin resistance and BMI, but occurs at an older age.
Detection of GAD‐Reactive CD4+ Cells in So‐Called “Type 1B” Diabetes
Authors: SHIMADA AKIRA , KODAMA KEIICHI , MORIMOTO JIRO , OIKAWA YOICHI , IRIE JUNICHIRO , NAKAGAWA YOSHINORI , MATSUBARA KOICHI , MARUYAMA TARO , SARUTA TAKAO
Published: January 2004

Abstract: Objective—Although the majority of type 1 diabetes is considered to be type 1A, some patients with type 1 diabetes have no islet‐associated autoantibody in their serum. This type of type 1 diabetes has usually been diagnosed as type 1B on the basis of islet‐associated autoantibody‐negativity. In this study, we tried to demonstrate the existence of islet‐associated antigen‐specific T cells in type 1 diabetes without islet‐associated autoantibody. Research Design and Methods—Peripheral blood samples were obtained from 110 Japanese diabetic patients, including 15 type 2 diabetic patients. Measurement of islet‐associated antigen‐specific cytokine response was performed by intracellular cytokine staining for flow cytometry. Results—The number of GAD‐reactive IFN‐γ‐producing CD4+ cells in 50,000 CD4+ cells in diabetics with type 1B (113.6 ± 34.6, median 45), type 1A (132.4 ± 33.3, median 25), and LADA (154.4 ± 44.1, median 20) was higher than that in type 2 diabetics (0.3 ± 0.3, median 0) and control subjects (3.8 ± 2.4, median 0). When the normal upper limit of the number of GAD‐reactive CD4+ cells was set at the mean + 3SD of values in control subjects, at least half (52.4%) of the so‐called “type 1B” patients were positive for GAD‐reactive IFN‐γ‐producing CD4+ cells, a significantly larger proportion than that in type 2 diabetics (0%; p < 0.001). Conclusions—Assessment of T cell reactivity against islet‐associated antigen may contribute to the diagnosis of “autoimmune‐related” type 1 diabetes.
Autoantibodies to GAD65 and IA‐2 Antibodies Are Increased, but Not Tissue Transglutaminase (TTG‐Ab) in Type 2 Diabetes Mellitus (T2DM) Patients from South India
Authors: SANJEEVI C. B. , BALAJI M. , BALAJI V. , SESHIAH V.
Published: January 2004

Abstract: The frequency of autoantibody‐positve T2DM patients identified as LADA was shown to be high in T2DM patients from Eastern India. In this study, we measured autoantibodies to GAD65, IA‐2, and TTG in T2DM patients from Southern India. Our results show that either GAD65 or IA‐2 was present in 30% of T2DM patients (n= 155) and in 3% of controls for GAD65 and 1% of controls for IA‐2 (n= 105). TTG‐Ab was not increased in T2DM patients compared to controls. Our results suggest that LADA is more frequent in Southern Indians than in European Caucasians, but not as high as in Eastern Indians.
IA‐2 Autoantibodies Are Predominant in Latent Autoimmune Diabetes in Adults Patients from Eastern India
Authors: KANUNGO ALOK , SANJEEVI C. B.
Published: January 2004

Abstract: Autoimmune diabetes or latent autoimmune diabetes in adults (LADA) among the clinically diagnosed type 2 diabetes patients from Cuttack in Eastern India was studied. GAD65 and IA‐2 autoantibodies were measured by radioligand binding assay using recombinant human GAD65 and IA‐2. The frequency of GAD65 was not significantly different between patients and controls. However, IA‐2 antibodies were predominant in LADA patients and there were two distinct peaks, one in the age group of 20 to 30 years and another in the age group of 50 to 60 years. The data suggest that LADA is more frequent in Eastern Indian T2DM patients and the IA‐2 is the predominant autoantibody in this population.
Parents Want to Know if Their Child Is at High Risk of Getting Diabetes
Authors: STOLT U. GUSTAFSSON , LISS P‐E. , LUDVIGSSON J. ,   ABIS STUDY GROUP
Published: January 2004

Abstract: Not least among professionals, voices have been raised against screening research projects, which have been regarded as involving a risk of being unethical as they may disturb, scare, or even harm the included people without giving enough benefit. This problem with large‐scale screening should be especially pronounced if increased risk of a serious disease like type 1 diabetes is identified when no effective prevention is available, and even more problematic if children were involved. ABIS (All Babies in Southeast Sweden) is a screening project including 17,000 newborn babies in the general population, followed prospectively to identify children at risk to get diabetes, and to study the influence of environmental factors causing the disease process. Four hundred randomly selected ABIS families received a questionnaire on attitudes and ethical questions regarding the project to be answered anonymously: 293/400 (73.3%) answered; 279/293 (95.3%) stated that they regarded it their right to be informed of results in the study and 278/293 (94.9%) said they really want to know. In fact, 254/293 (86.7%) report wanting to know if their child has increased risk of getting diabetes even if there is no preventive measure available. This clear result supports the view that this type of study may well be ethically justified as long as informed consent can be given based on adequate understanding and voluntariness. The results may have implications for the design of future screening studies.