Itoh N, Hanafusa T, Yamagata K, Nakajima H, Tomita K, Tamura S, Inada M, Kawata S, Kono N, Kuwajima M
Second Department of Internal Medicine, Osaka University Medical School, Japan.
Diabetologia. 1995 Jun;38(6):667-71. doi: 10.1007/BF00401837.
Viral infection is assumed to trigger or exacerbate autoimmune responses against pancreatic beta cells leading to the development of insulin-dependent diabetes mellitus (IDDM). We therefore examined by polymerase chain reaction the presence of two candidate viruses, cytomegalovirus and Epstein-Barr virus, in IDDM pancreases. Pancreas tissues were obtained by biopsy under laparoscopy from 16 recent-onset IDDM patients: age 17-53 years; disease duration 0-7 months; six had flu-like symptoms before onset. Frozen sections were made and subjected to DNA amplification. DNA samples were prepared from the frozen sections and polymerase chain reaction was performed using primers specific to cytomegalovirus, Epstein-Barr virus and control gene for HLA-DP. Cytomegalovirus- and Epstein-Barr virus-infected cells were used for positive control. Southern blot analysis could detect cytomegalovirus DNA from as few as 2 x 10(-1) cytomegalovirus-infected cells and Epstein-Barr virus DNA from two Epstein-Barr virus-infected cells. This highly sensitive analysis, however, could not detect cytomegalovirus or Epstein-Barr virus genomes in pancreases of recent-onset IDDM. A single copy human gene (HLA-DP) was amplified from all IDDM pancreases indicating that DNA amplification was performed without inhibition. We conclude that cytomegalovirus or Epstein-Barr virus genomes are unlikely to exist in pancreas biopsy specimens of recent-onset IDDM patients.
病毒感染被认为会引发或加剧针对胰腺β细胞的自身免疫反应,从而导致胰岛素依赖型糖尿病(IDDM)的发生。因此,我们通过聚合酶链反应检测了IDDM胰腺中两种候选病毒——巨细胞病毒和爱泼斯坦-巴尔病毒的存在情况。通过腹腔镜活检从16例近期发病的IDDM患者获取胰腺组织:年龄17 - 53岁;病程0 - 7个月;6例发病前有流感样症状。制作冰冻切片并进行DNA扩增。从冰冻切片制备DNA样本,并使用针对巨细胞病毒、爱泼斯坦-巴尔病毒和HLA - DP对照基因的引物进行聚合酶链反应。使用感染巨细胞病毒和爱泼斯坦-巴尔病毒的细胞作为阳性对照。Southern印迹分析可从低至2×10⁻¹个感染巨细胞病毒的细胞中检测到巨细胞病毒DNA,从两个感染爱泼斯坦-巴尔病毒的细胞中检测到爱泼斯坦-巴尔病毒DNA。然而,这种高灵敏度分析未能在近期发病的IDDM胰腺中检测到巨细胞病毒或爱泼斯坦-巴尔病毒基因组。从所有IDDM胰腺中均扩增出单拷贝人类基因(HLA - DP),表明DNA扩增未受抑制。我们得出结论,近期发病的IDDM患者胰腺活检标本中不太可能存在巨细胞病毒或爱泼斯坦-巴尔病毒基因组。