PreprintArticleVersion 1Preserved in Portico This version is not peer-reviewed
Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus
Version 1
: Received: 3 February 2021 / Approved: 5 February 2021 / Online: 5 February 2021 (11:34:57 CET)
How to cite:
Jimbo, E.; Kobayashi, T.; Takeshita, A.; Mine, K.; Nagafuchi, S.; Fukui, T.; Yagihashi, S. Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus. Preprints2021, 2021020159. https://doi.org/10.20944/preprints202102.0159.v1
Jimbo, E.; Kobayashi, T.; Takeshita, A.; Mine, K.; Nagafuchi, S.; Fukui, T.; Yagihashi, S. Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus. Preprints 2021, 2021020159. https://doi.org/10.20944/preprints202102.0159.v1
Jimbo, E.; Kobayashi, T.; Takeshita, A.; Mine, K.; Nagafuchi, S.; Fukui, T.; Yagihashi, S. Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus. Preprints2021, 2021020159. https://doi.org/10.20944/preprints202102.0159.v1
APA Style
Jimbo, E., Kobayashi, T., Takeshita, A., Mine, K., Nagafuchi, S., Fukui, T., & Yagihashi, S. (2021). Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus. Preprints. https://doi.org/10.20944/preprints202102.0159.v1
Chicago/Turabian Style
Jimbo, E., Tomoyasu Fukui and Soroku Yagihashi. 2021 "Immunohistochemical Detection of Enteroviruses in Pancreatic Tissues of Patients with Type 1 Diabetes Using a Polyclonal Antibody Against 2A Protease of Coxsackievirus" Preprints. https://doi.org/10.20944/preprints202102.0159.v1
Abstract
The need for antiserum for immunohistochemical (IHC) detection of enterovirus (EV) in formaldehyde fixed and paraffin-embedded (FFPE) specimens is increasing. The standard monoclonal antibody against EV-envelope protein (VP1), clone 5D8/1, was proven to cross-react with other proteins. Another candidate marker of EV proteins is 2A protease (2Apro), which is coded by the EV gene and translated by host cells during EV replication. We raised polyclonal antiserum by immunizing rabbits with an 18-mer peptide of Coxsackievirus B1 (CVB1)-2A protease (2Apro) and examined the specificity and sensitivity for EV on FFPE tissue samples. ELISA study showed a high titer of antibody for CVB1-2Apro. IHC demonstrated that antiserum against 2Apro reacted with CVB1-infected Vero-cells. Confocal microscopy demonstrated that 2Apro labelled by the antibody located in the same cell with VP1 stained with 5D8/1. IHC demonstrated dense positive reactions pancreatic islets of EV-associated fulminant type 1 diabetes (FT1DM), and located in the same cell stained positive with 5D8/1. Specificity of IHC staining FT1DM pancreas was confirmed by absorption with an excessive concentration of immunized peptide. In conclusion, our study provides a new polyclonal antiserum against CVB1 2Apro which may be useful for detection of EV-infected human tissues stored as archive of FFPE tissue samples.
Keywords
Enterovirus; Coxsackievirus; 2A protease; polyclonal antibody; type 1 diabetes
Subject
Biology and Life Sciences, Anatomy and Physiology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
