Detection of enterovirus in the islet cells of patients with type 1 diabetes: what do we learn from immunohistochemistry?

Immunohistochemical detection of enteroviruses in pancreatic tissues of patients with type 1 diabetes using a polyclonal antibody against 2A protease of Coxsackievirus

“…Busse et al 20 reported a new method to detect EV-RNA using short fluorescently labeled oligonucleotide. In addition, it has been reported that the monoclonal antibody against EV-VP1 protein (5D8/1) cross-reacts with other proteins, including a component of the mitochondrial adenosine triphosphate synthase (ATP5B) and an isoform of creatine kinase in human organs 7 and requires optimized IHC conditions 8,9 . We demonstrated that a polyclonal antibody against CVB1-2A pro showed positive staining in formalin-fixed Vero cells infected by CVB1 and the FFPE pancreatic tissues of EV-induced fulminant type 1 diabetes in the present study.…”

“…However, no polyclonal or monoclonal antibodies against 2A pro of EV are yet to be commercially available for IHC analysis 6 . In addition, the standard antiserum against EV-capsid protein 1 (VP1) often suffers from problematic cross-reactions with other protein epitopes 7,8 . Thus, the establishment of optimized conditions that preclude cross-reactions with non-target protein epitopes is required in IHC analysis [7][8][9] .…”

“…In addition, the standard antiserum against EV-capsid protein 1 (VP1) often suffers from problematic cross-reactions with other protein epitopes 7,8 . Thus, the establishment of optimized conditions that preclude cross-reactions with non-target protein epitopes is required in IHC analysis [7][8][9] . For the identification of microbes, it is often difficult to obtain positive results in situ hybridization, because EV-RNA, especially in pancreatic tissues, is likely degraded by many digestive enzymes over the prolonged post-mortem conditions before fixation of the samples 10 .…”

Application of bioinformatics in probe design enables detection of enteroviruses on different taxonomic levels by advanced in situ hybridization technology

“…Thus far, immunohistochemistry-based studies on EVs have mostly been based on a single commercially available antibody clone 5D8/1 (Dako, Glostrup, Denmark) that recognizes viral capsid protein VP1 5 [12]. This method has provoked discussions with regards to its sensitivity and specificity in recent publications [13][14][15][16]. Combining viral protein findings with standardized and reliable in situ hybridization 6 (ISH) mRNA results would strengthen the reliability of viral detection.…”

An ancestral retroviral protein identified as a therapeutic target in type-1 diabetes

“…Most particularly, enteroviruses have been associated with T1D in many instances, but this still raises questions (7)(8)(9)(10)(11). However, a meta-analysis based on molecular studies showed a significant association between enteroviral infection and T1D (12).…”