Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection

Abstract The generation of a human pancreatic beta cell line which reproduces the responses seen in primary beta cells, but is amenable to propagation in culture, has long been an important goal in diabetes research. This is particularly true for studies focussing on the role of enteroviral infectio...

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Autores principales:	Jessica R. Chaffey, Jay Young, Kaiyven A. Leslie, Katie Partridge, Pouria Akhbari, Shalinee Dhayal, Jessica L. Hill, Kyle C. A. Wedgwood, Edward Burnett, Mark A. Russell, Sarah J. Richardson, Noel G. Morgan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2b039b3f52724d06b5086c5235906f58
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spelling	oai:doaj.org-article:2b039b3f52724d06b5086c5235906f582021-12-02T17:06:24ZInvestigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection10.1038/s41598-021-94878-y2045-2322https://doaj.org/article/2b039b3f52724d06b5086c5235906f582021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94878-yhttps://doaj.org/toc/2045-2322Abstract The generation of a human pancreatic beta cell line which reproduces the responses seen in primary beta cells, but is amenable to propagation in culture, has long been an important goal in diabetes research. This is particularly true for studies focussing on the role of enteroviral infection as a potential cause of beta-cell autoimmunity in type 1 diabetes. In the present work we made use of a clonal beta cell line (1.1B4) available from the European Collection of Authenticated Cell Cultures, which had been generated by the fusion of primary human beta-cells with a pancreatic ductal carcinoma cell, PANC-1. Our goal was to study the factors allowing the development and persistence of a chronic enteroviral infection in human beta-cells. Since PANC-1 cells have been reported to support persistent enteroviral infection, the hybrid 1.1B4 cells appeared to offer an ideal vehicle for our studies. In support of this, infection of the cells with a Coxsackie virus isolated originally from the pancreas of a child with type 1 diabetes, CVB4.E2, at a low multiplicity of infection, resulted in the development of a state of persistent infection. Investigation of the molecular mechanisms suggested that this response was facilitated by a number of unexpected outcomes including an apparent failure of the cells to up-regulate certain anti-viral response gene products in response to interferons. However, more detailed exploration revealed that this lack of response was restricted to molecular targets that were either activated by, or detected with, human-selective reagents. By contrast, and to our surprise, the cells were much more responsive to rodent-selective reagents. Using multiple approaches, we then established that populations of 1.1B4 cells are not homogeneous but that they contain a mixture of rodent and human cells. This was true both of our own cell stocks and those held by the European Collection of Authenticated Cell Cultures. In view of this unexpected finding, we developed a strategy to harvest, isolate and expand single cell clones from the heterogeneous population, which allowed us to establish colonies of 1.1B4 cells that were uniquely human (h1.1.B4). However, extensive analysis of the gene expression profiles, immunoreactive insulin content, regulated secretory pathways and the electrophysiological properties of these cells demonstrated that they did not retain the principal characteristics expected of human beta cells. Our data suggest that stocks of 1.1B4 cells should be evaluated carefully prior to their use as a model human beta-cell since they may not retain the phenotype expected of human beta-cells.Jessica R. ChaffeyJay YoungKaiyven A. LeslieKatie PartridgePouria AkhbariShalinee DhayalJessica L. HillKyle C. A. WedgwoodEdward BurnettMark A. RussellSarah J. RichardsonNoel G. MorganNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jessica R. Chaffey Jay Young Kaiyven A. Leslie Katie Partridge Pouria Akhbari Shalinee Dhayal Jessica L. Hill Kyle C. A. Wedgwood Edward Burnett Mark A. Russell Sarah J. Richardson Noel G. Morgan Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
description	Abstract The generation of a human pancreatic beta cell line which reproduces the responses seen in primary beta cells, but is amenable to propagation in culture, has long been an important goal in diabetes research. This is particularly true for studies focussing on the role of enteroviral infection as a potential cause of beta-cell autoimmunity in type 1 diabetes. In the present work we made use of a clonal beta cell line (1.1B4) available from the European Collection of Authenticated Cell Cultures, which had been generated by the fusion of primary human beta-cells with a pancreatic ductal carcinoma cell, PANC-1. Our goal was to study the factors allowing the development and persistence of a chronic enteroviral infection in human beta-cells. Since PANC-1 cells have been reported to support persistent enteroviral infection, the hybrid 1.1B4 cells appeared to offer an ideal vehicle for our studies. In support of this, infection of the cells with a Coxsackie virus isolated originally from the pancreas of a child with type 1 diabetes, CVB4.E2, at a low multiplicity of infection, resulted in the development of a state of persistent infection. Investigation of the molecular mechanisms suggested that this response was facilitated by a number of unexpected outcomes including an apparent failure of the cells to up-regulate certain anti-viral response gene products in response to interferons. However, more detailed exploration revealed that this lack of response was restricted to molecular targets that were either activated by, or detected with, human-selective reagents. By contrast, and to our surprise, the cells were much more responsive to rodent-selective reagents. Using multiple approaches, we then established that populations of 1.1B4 cells are not homogeneous but that they contain a mixture of rodent and human cells. This was true both of our own cell stocks and those held by the European Collection of Authenticated Cell Cultures. In view of this unexpected finding, we developed a strategy to harvest, isolate and expand single cell clones from the heterogeneous population, which allowed us to establish colonies of 1.1B4 cells that were uniquely human (h1.1.B4). However, extensive analysis of the gene expression profiles, immunoreactive insulin content, regulated secretory pathways and the electrophysiological properties of these cells demonstrated that they did not retain the principal characteristics expected of human beta cells. Our data suggest that stocks of 1.1B4 cells should be evaluated carefully prior to their use as a model human beta-cell since they may not retain the phenotype expected of human beta-cells.
format	article
author	Jessica R. Chaffey Jay Young Kaiyven A. Leslie Katie Partridge Pouria Akhbari Shalinee Dhayal Jessica L. Hill Kyle C. A. Wedgwood Edward Burnett Mark A. Russell Sarah J. Richardson Noel G. Morgan
author_facet	Jessica R. Chaffey Jay Young Kaiyven A. Leslie Katie Partridge Pouria Akhbari Shalinee Dhayal Jessica L. Hill Kyle C. A. Wedgwood Edward Burnett Mark A. Russell Sarah J. Richardson Noel G. Morgan
author_sort	Jessica R. Chaffey
title	Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
title_short	Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
title_full	Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
title_fullStr	Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
title_full_unstemmed	Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection
title_sort	investigation of the utility of the 1.1b4 cell as a model human beta cell line for study of persistent enteroviral infection
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/2b039b3f52724d06b5086c5235906f58
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Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection

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