Kjersti S. Rønningen

Susceptibility to human type 1 diabetes at IDDM2 is determined by tandem repeat variation at the insulin gene minisatellite locus

The IDDM2 locus encoding susceptibility to type 1 diabetes was mapped previously to a 4.1–kb region spanning the insulin gene and a minisatellite or variable number of tandem repeats (VNTR) locus on human chromosome 11p15.5. By ‘cross–match’ haplotype analysis and linkage disequilibrium mapping, we have mapped the mutation IDDM2 to within the VNTR itself. Other polymorphisms were systematically excluded as primary disease determinants. Transmission of IDDM2 may be influenced by parent–of–origin phenomena. Although we show that the insulin gene is expressed biallelically in the adult pancreas, we present preliminary evidence that the level of transcription in vivo is correlated with allelic variation within the VNTR. Allelic variation at VNTRs may play an important general role in human disease.

HLA-DQA1 and HLA-DQB1 genes may jointly determine susceptibility to develop multiple sclerosis

Serologic DR typing and genomic DRB1, DQA1, DQB1, DPA1, and DPB1 typing using sequence-specific oligonucleotides were performed in 69 multiple sclerosis (MS) patients and 181 healthy controls on in vitro amplified DNA. The frequencies of DR2 as well as the DR2-associated DQA1*0102 and DQB1*0602 alleles were increased whereas DR7 was decreased among MS patients. The distribution of DR4 subtypes as well as DP alleles were similar in patients and healthy controls. All but one of 23 DR4-positive MS patients carried the DQB1*0302 allele, whereas five of five DR7-positive MS patients carried the DQB1*0303 allele. Of the MS patients, 99% compared to 79% of the controls carried DQA1 alleles encoding glutamine at residue 34, while 97% of the MS patients compared to 72% of the controls carried DQB1 alleles encoding DQ beta chains sharing long polymorphic stretches. A combination of such DQA1 and DQB1 alleles was carried by 96% of the MS patients and 60% of the controls, suggesting an association between MS and a combination of particular DQA1 alleles and DQB1 alleles. The corresponding DQ alpha beta heterodimers may have in common an ability to bind a particular peptide.

The amino acid at position 57 of the HLA-DQB chain and susceptibility to develop insulin-dependent diabetes mellitus

In Caucasoids HLA-DQB1 genes encoding amino acids other than aspartic acid at position 57 of the DQ beta chain (non-Asp-57) are associated with susceptibility to develop insulin-dependent diabetes mellitus (IDDM), while resistance is associated with aspartic acid at this residue (Asp-57). Following amplification of genomic DNA by the polymerase chain reaction, the DQB1 alleles of 87 random Norwegian IDDM patients and 187 healthy controls were investigated with 11 different sequence-specific oligonucleotide probes. Of these patients 82% carried DQB1 alleles encoding non-Asp-57 at both of their DQ beta chains, compared to 27% of the controls (relative risk = 12.2, p less than 0.0001). Sixteen percent of the patients (versus 51% of the controls) were heterozygous Asp-57/non-Asp-57. Two percent of the patients (22% of the controls) were apparently Asp-57 homozygous. The results demonstrate that non-Asp-57 DQ beta chains are associated with susceptibility to develop IDDM but also indicate that the protection associated with DQ beta Asp-57 may not be as dominant as reported by others.

Polymorphic analysis of the human MHC-linked heat shock protein 70 (HSP70-2) and HSP70-Hom genes in insulin-dependent diabetes mellitus (IDDM)

In the present study we characterized the frequencies of two polymorphisms within the MHC‐linked heat shock protein (HSP) 70 genes in patients with insulin‐dependent diabetes mellitus (IDDM) (n= 114) and healthy control individuals (n = 110). Significant differences in genotype and allelic frequencies were observed for both polymorphisms between randomly selected patients and controls. However, for the HSP70‐2 polymorphisms this was solely due to linkage disequilibrium with DR3. The rare HSP70‐Hom 2‐allele was significantly more frequent in controls than in patients. It showed strong association with certain tumour necrosis factor (TNF) (class III) and HLA‐B and ‐A (class I) alleles independent of HLA‐DQ and ‐DR alleles. By typing 257 individuals from 55 IDDM multiple‐case families two extended MHC‐haplotypes, including class II‐, TNF‐ and class I‐markers, carrying the rare HSP70‐Hom allele were defined. One was only transmitted to diabetic offspring, whereas the other was only transmitted to unaffected offspring. The functional implication of the polymorphism in the heat shock‐inducible HSP70‐2 gene was analysed by studying HSP70‐2 mRNA expression after heat shock in peripheral blood mononuclear cells from individuals with different HSP70‐2 genotypes. Preliminary data showed that individuals homozygous for the PstI 8.5‐kb allele consistently had slightly lower expression than heterozygous and 9·0‐kb homozygous individuals.

Distribution of HLA class II alleles among Norwegian caucasians

We report genomic HLA class II typing of 181 randomly selected Norwegian controls. Seventeen DRB1, 7 DQA1, 10 DQB1, 2 DPA1, and 16 DPB1 alleles were found in the tested population. HLA class II antigen and allele frequencies are given, as well as the distribution of DRB1, DQA1, DQB1 haplotypes. Linkage disequilibrium between some DPB1 alleles and DRB1 and/or DQB1 alleles are also reported.

Longitudinal observation of parechovirus in stool samples from Norwegian infants

Parechoviruses are assumed to be common infectious agents, but their epidemiologic and pathogenic properties are not well known. The aim of the present study was to assess the prevalence and molecular epidemiology of Parechovirus in Norwegian infants, as well as to investigate whether the presence of virus correlated with symptoms of infection. A group of 102 infants was longitudinally followed: 51 infants with a high genetic risk for type 1 diabetes (aged 3–35 months), and 51 children without this genotype (aged 3–12). Stool samples were obtained each month, and symptoms of infection were recorded regularly on questionnaires. Human parechovirus was detected in 11.3% of 1,941 samples examined by real‐time RT‐PCR. There was a distinct seasonality, peaking from September to December. By 12 months of age, 43% of the infants had had at least one infection, while 86% of the infants had encountered the virus by the end of the second year. Based on the VP1 sequence, human parechovirus 1 was the most prevalent type (76%), followed by human parechovirus 3 (13%), human parechovirus 6 (9%), an unclassified human parechovirus (1%), and human parechovirus 2 (1%). Ljungan virus, a murine parechovirus, was examined with a separate real‐time RT‐PCR, but no virus was detected. There was no significant association between infections and the following symptoms: coughing, sneezing, fever, diarrhea or vomiting. In conclusion, human parechovirus infects frequently infants at an early age without causing disease. J. Med. Virol. 80:1835–1842, 2008.

Insulin gene region-encoded susceptibility to IDDM maps upstream of the insulin gene.

The gene region on chromosome 11p15.5 known to be involved in insulin-dependent diabetes mellitus (IDDM) susceptibility was recently mapped to a 4.1-kilobase region including the insulin gene. The region contains 10 candidate polymorphisms that are in strong linkage disequilibrium. By genotyping 7 of these 10 polymorphisms and the tyrosine hydroxylase microsatellite in Finnish Caucasoid IDDM patients and control subjects, we demonstrate that many of the polymorphisms found to be associated with IDDM in other Caucasoid populations do not show any association in this Finnish population. Of the polymorphisms typed, only those at –23 Hph I and the variable number of tandem repeats (VNTR) sites confer significant relative risk. Furthermore, we have demonstrated that the –23 Hph I polymorphism cannot explain the association. Comparison of the genotypic patterns observed here and previously suggests that the VNTR is the most likely candidate for IDDM2. The VNTR is located adjacent to defined regulatory DNA sequences affecting insulin gene expression, which suggests a possible effect on expression of insulin or one of the neighboring genes, tyrosine hydroxylase or insulin-like growth factor 2.

High Prevalence of Human Enterovirus A Infections in Natural Circulation of Human Enteroviruses

ABSTRACT Human enterovirus (HEV) infections can be asymptomatic or cause only mild illness; recent evidence may implicate HEV infection in type 1 diabetes mellitus and myocarditis. Here, we report the molecular characterization of HEV obtained in serial monthly collections from healthy Norwegian infants. A total of 1,255 fecal samples were collected from 113 healthy infants beginning at age 3 months and continuing to 28 months. The samples were analyzed for HEV nucleic acid by real-time PCR. Fifty-eight children (51.3%) had HEV infections. One hundred forty-five positive samples were typed directly by nucleotide sequencing of the VP1 region. HEV-A was detected most frequently, with an overall prevalence of 6.8%. HEV-B was present in 4.8% of the samples and HEV-C in only 0.2% of the samples. No poliovirus or HEV-D group viruses were detected. Twenty-two different serotypes were detected in the study period: the most common were EV71 (14.5%), CAV6 (10.5%), CAV4 (8.9%), E18 (8.9%), and CBV3 (7.3%). These findings suggest that the prevalence of HEV infections in general, and HEV-A infections in particular, has been underestimated in epidemiological studies based on virus culture.

Assessing the validity of the association between the SUMO4 M55V variant and risk of type 1 diabetes

Assessing the validity of the association between the SUMO4 M55V variant and risk of type 1 diabetes

Genetic Susceptibility Markers in Danish Patients with Type 1 (Insulin-Dependent) Diabetes-Evidence for Polygenecity in Man

Fifty-five Danish families with two offspring concordant for type 1 diabetes-identified through a nationwide population-based survey, and 57 “true sporadic” cases-matched with familial cases for age at onset, but with no IDDM-affected first-degree relatives and long disease duration, and 110 control subjects were typed for putative genetic susceptibility markers for type 1 diabetes identified from a pathogenetic model. The markers included MHC class I, II and III loci, the manganese superoxide dismutase (MnSOD) locus (chr. 6q), interleukin-1β (IL1B), the IL-1 receptor antagonist (IL1RN), and the IL-1 type 1 receptor (IL1RI) loci (each chr. 2q). No significant differences between familial and sporadic cases were found within the MHC region (including the following loci: HLA-DQ,-DR, heat shock protein (HSP) 70, tumour necrosis factor (TNF), HLA-B and-A). In both groups of patients 11% were negative for both DQA1*0301-DQB 1*0302 and DQA1*0501-DQB1*0201 genotypes, and 7% of the type 1 diabetics had genotypes ...