Enterovirus RNA Is Found in Peripheral Blood Mononuclear Cells in a Majority of Type 1 Diabetic Children at Onset

• Published in: Diabetes (New York, N.Y.) Vol. 51; no. 6; pp. 1964 - 1971
• Main Authors: Yin, Hong, Berg, Anna-Karin, Tuvemo, Torsten, Frisk, Gun
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Diabetes Association 01.06.2002
• Subjects: Adolescent; Alleles; Antibodies, Viral - blood; Antigens; Autoantibodies - blood; Biological and medical sciences; Causes of; Child; Child, Preschool; Diabetes; Diabetes in children; Diabetes Mellitus, Type 1 - genetics; Diabetes Mellitus, Type 1 - immunology; Diabetes Mellitus, Type 1 - virology; Diabetes. Impaired glucose tolerance; Disease susceptibility; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Enterovirus - genetics; Enterovirus - immunology; Environmental aspects; Etiopathogenesis. Screening. Investigations. Target tissue resistance; Female; Genetic Predisposition to Disease; Glutamate Decarboxylase - immunology; HLA-DR Antigens - genetics; HLA-DRB1 Chains; Humans; Immunoglobulin M - blood; Infant; Infections; Insulin; Isoenzymes - immunology; Juvenile diabetes; Leukocytes, Mononuclear - virology; Male; Medical sciences; MEDICIN; MEDICINE; Nuclear Family; Phylogenetics; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - blood; Sequence Analysis; Siblings; Type 1 diabetes; Viral infections; Viruses; Sweden; Finland; Endocrinopathy; Human; Virus; Immunopathology; Blood cell; Mononuclear cell; RNA; Picornaviridae; Insulin dependent diabetes; Autoimmune disease; Enterovirus; Child
• ISSN: 0012-1797; 1939-327X; 1939-327X
• DOI: 10.2337/diabetes.51.6.1964

Enterovirus RNA Is Found in Peripheral Blood Mononuclear Cells in a Majority of Type 1 Diabetic Children at Onset Hong Yin , Anna-Karin Berg , Torsten Tuvemo and Gun Frisk From the Department of Women’s and Children’s Health, Uppsala University, Akademiska Hospital, 751 85 Uppsala, Sweden Abstract We have studied the occurrence of enterovirus (EV)-RNA at the onset of childhood type 1 diabetes in all 24 new cases of childhood type 1 diabetes during 1 year in Uppsala county, Sweden. We also studied 24 matched control subjects and 20 siblings of the patients. RNA was isolated from peripheral blood mononuclear cells and EV-RNA detected by RT-PCR. Primers (groups A and B) corresponding to conserved regions in the 5′ noncoding region (NCR) of EV were used in the PCRs, and the amplicons were sequenced. By the use of group A primers, EV-RNA was found in 12 (50%) of the 24 type 1 diabetic children, 5 (26%) of 19 siblings, and none of the control subjects. Both patients and siblings showed a higher frequency of EV-RNA compared with the control subjects. The group B primers detected EV-RNA in all three groups but did not show statistically significant differences between the groups. The EV-RNA positivity with the group B primers was 11 (46%) of 24 in the type 1 diabetic children, 11 (58%) of 19 in the siblings, and 7 (29%) of 24 in the control subjects. The significant difference between groups seen with the group A primers but not with the group B primers might indicate the existence of diabetogenic EV strains. The phylogenetic analysis of the PCR products revealed clustering of the sequences from patients and siblings into five major branches when the group A PCR primers were used. With the group B primers, the sequences from patients, siblings, and control subjects formed three major branches in the phylogenetic tree, where 6 of the 7 control subjects clustered together in a sub-branch of CBV-4/VD2921. Seven of the type 1 diabetic children clustered together in another sub-branch of CBV-4/VD2921. Five of the type 1 diabetic children formed a branch together with the CBV-4/E2 strain, four clustered together with CBV-5, and one formed a branch with echovirus serotype. The presence of EV-RNA in the blood cells of most newly diagnosed type 1 diabetic children supports the hypothesis that a viral infection acts as an exogenous factor. In addition, sequencing of the PCR amplicons from the type 1 diabetic children, their siblings, and matched control subjects might reveal differences related to diabetogenic properties of such a virus. Footnotes Address correspondence and reprint requests to Gun Frisk, Department of Women’s and Children’s Health, Uppsala University, Akademiska Hospital, 751 85 Uppsala, Sweden. E-mail: gun.frisk{at}kbh.uu.se . Received for publication 24 May 2001 and accepted in revised form 20 February 2002. CBV, Coxsackievirus B; CPE, cytopathic effect; EMEM, Eagle’s minimum essential medium; EV, enterovirus; IAA, insulin autoantibody; ICA, islet cell antibody; NCR, noncoding region; NT, neutralizing antibody; PBMC, peripheral blood mononuclear cell; RT, reverse transcription. DIABETES