Investigation of the utility of the 1.1B4 cell as a model human beta cell line for study of persistent enteroviral infection.
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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DOI：
10.1038/s41598-021-94878-y
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发表时间：
2021-08-02
影响因子：
4.6
通讯作者：
Morgan NG
中科院分区：
文献类型：
--
作者：
Chaffey JR;Young J;Leslie KA;Partridge K;Akhbari P;Dhayal S;Hill JL;Wedgwood KCA;Burnett E;Russell MA;Richardson SJ;Morgan NG
长期以来，糖尿病研究的一个重要目标是建立一种人胰腺β细胞系，它可以复制出在原代β细胞中看到的反应，但可以在培养中繁殖。这一点在关注肠道病毒感染作为1型糖尿病患者β细胞自身免疫的潜在原因的研究中尤其如此。在目前的工作中，我们利用了来自欧洲鉴定细胞培养集合的克隆β细胞系(1.1B4)，该克隆细胞系是由原代人类β细胞与胰腺导管癌细胞PANC-1融合而产生的。我们的目标是研究允许慢性肠道病毒感染在人类β细胞中发展和持续的因素。由于PANC-1细胞被报道支持持续的肠道病毒感染，杂交的1.1B4细胞似乎为我们的研究提供了一个理想的载体。为了支持这一点，用一种最初从1型糖尿病儿童的胰腺分离的柯萨奇病毒CVB4.E2感染细胞，感染的复数很低，导致了一种持续感染的状态。对分子机制的研究表明，这种反应是由一些意想不到的结果促进的，包括细胞明显未能上调某些抗病毒反应基因产物对干扰素的反应。然而，更详细的探索表明，这种反应的缺乏仅限于被人类选择性试剂激活或检测到的分子靶标。相比之下，令我们惊讶的是，这些细胞对啮齿动物选择性试剂的反应要好得多。然后，通过多种方法，我们确定了1.1B4细胞的群体并不是同质的，而是含有啮齿动物和人类细胞的混合物。无论是我们自己的细胞库，还是欧洲鉴定细胞培养集合会持有的细胞库，都是如此。鉴于这一意想不到的发现，我们开发了一种策略，从异质群体中收集、分离和扩大单细胞克隆，这使得我们能够建立人类独有的1.1B4细胞克隆(h1.1.B4)。然而，对这些细胞的基因表达谱、免疫活性胰岛素含量、调节的分泌途径和电生理特性的广泛分析表明，它们并不保留人类β细胞的主要特征。我们的数据表明，1.1B4细胞的存量在用作模型人类β细胞之前应该仔细评估，因为它们可能不会保留人类β细胞所期望的表型。
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.
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影响因子：
4.5
作者：
Akhbari P;Richardson SJ;Morgan NG
通讯作者：
Morgan NG
早期和童年时期的病毒素以及胰岛自身免疫性和1型糖尿病的发展：观察性研究的系统评价和荟萃分析。
影响因子：
11.1
作者：
Faulkner CL;Luo YX;Isaacs S;Rawlinson WD;Craig ME;Kim KW
通讯作者：
Kim KW
影响因子：
7.7
作者：
Lundberg, Marcus;Krogvold, Lars;Skog, Oskar
通讯作者：
Skog, Oskar
影响因子：
8.1
作者：
Benazra, Marion;Lecomte, Marie-Jose;Ravassard, Philippe
通讯作者：
Ravassard, Philippe
影响因子：
4.8
作者：
MacFarlane, WM;Chapman, JC;Dunne, MJ
通讯作者：
Dunne, MJ