A chicken primary B cell culture model to study the pathogenesis and improve the control of immunosuppressive viruses of poultry

鸡原代 B 细胞培养模型，用于研究发病机制并提高家禽免疫抑制病毒的控制

• 批准号: NC/R001138/1
• 负责人: Andrew Broadbent
• 金额: $ 44.48万
• 依托单位: The Pirbright Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FR001138%2F1
• 关键词: chicken; primary; cell; culture; model

Chicken meat is an important source of dietary protein for people worldwide, and the poultry industry is important for securing enough food to feed the growing human population. Ensuring chickens remain healthy is therefore an economically important issue as well as a welfare issue for the birds. Chickens require a healthy immune system for vaccines to work effectively, and to fight off infection, so if their immune system is suppressed, birds will respond less well to vaccines and be more susceptible to catching infections, some of which can also infect people. The chicken immune system can become suppressed by infection with viruses that target a type of white blood cell known as a B cell. The main viruses that infect B cells in chickens are: Marek's disease virus (MDV), infectious bursal disease virus (IBDV), avian leukosis virus (ALV) and reticuloendotheliosis virus (REV). MDV and IBDV destroy B cells, whereas ALV and REV can cause the B cells to become cancerous and form a tumour. All these viruses can cause immunosuppression. These diseases are poorly controlled: New strains of IBDV have spread and vaccine failures have been reported, and ALV and REV continue to be isolated despite eradication efforts. There is therefore a need for new vaccines and control methods. It might be possible in the future to produce birds that are resistant to these diseases, but we need to better understand how the viruses interact with the cells they infect in order to realise this potential.Until recently, it was not possible to study how the chicken B cells responded to infection as the cells died when they were taken out of the birds. Therefore, most studies into how these viruses cause disease and immunosuppression had to be conducted using infected chickens that suffered from disease, which could be severe in some cases. Recently, it has become possible to keep chicken B cells alive in the laboratory once removed from birds, and one research group has already used the cells to study MDV infection. We have used the cells to study IBDV infection, and our data suggest that they would be a good model for studying how IBDV interacts with the bird immune system, without the need to infect birds directly. We believe the model has the potential to replace the use of infected birds, and reduce the total number of birds used in this kind of research. The aim of this project is to build on our previous data to establish the chicken B cell model for studying IBDV, and expand it to the study of ALV and REV. In order to do this, we have three objectives: First, we will confirm and validate the use of the model for studying IBDV by comprehensively quantifying changes in the level of gene expression in the B cells following infection compared to what happens in the infected bird. Second, we will expand the model to include ALV and REV. Third, we will use the cells to produce stocks of IBDV as, currently, virus stocks are produced by infecting birds and recovering the virus from organs, so if the B cells can be used for this purpose, it would replace the use of even more infected birds. We will also determine if the cells can be used to screen vaccines for how effective they are against locally circulating and new strains to improve control.Not only will we be able to replace and reduce the use of infected chickens, but using B cells is actually a better approach than using infected chickens to answer some scientifically important questions, as there is only one cell population as opposed to many, and it is possible to control the amount of virus the cells receive and to know for how long the cells have been infected. In addition, we can apply the model in the future to study how IBDV vaccines interact with the cells, and study co-infection with multiple viruses, so the project not only has a high 3Rs impact, but also a high scientific and translational impact.

鸡肉是全世界人民膳食蛋白质的重要来源，家禽业对于确保足够的食物来养活不断增长的人口非常重要。因此，确保鸡保持健康是一个重要的经济问题，也是一个鸟类福利问题。鸡需要一个健康的免疫系统才能让疫苗有效地发挥作用，并抵抗感染，所以如果它们的免疫系统受到抑制，鸟类对疫苗的反应就会降低，更容易感染，其中一些也会感染人类。鸡的免疫系统可能会受到病毒感染的抑制，这些病毒的目标是一种称为B细胞的白色血细胞。感染鸡B细胞的主要病毒有：马立克氏病病毒（MDV）、传染性法氏囊病病毒（IBDV）、禽白血病病毒（ALV）和网状内皮组织增生症病毒（REV）。MDV和IBDV破坏B细胞，而ALV和REV可导致B细胞癌变并形成肿瘤。所有这些病毒都能引起免疫抑制。这些疾病的控制很差：新的IBDV毒株已经传播，疫苗失败的报道已经出现，尽管有根除努力，ALV和REV仍然被分离出来。因此，需要新的疫苗和控制方法。将来也许有可能培育出对这些疾病有抵抗力的禽类，但我们需要更好地了解病毒如何与它们感染的细胞相互作用，以实现这一潜力。直到最近，人们才能研究鸡的B细胞如何对感染作出反应，因为细胞在从禽类体内取出时就死亡了。因此，大多数关于这些病毒如何引起疾病和免疫抑制的研究必须使用患有疾病的受感染鸡进行，在某些情况下可能是严重的。最近，一旦从鸟类中取出鸡B细胞，就可以在实验室中保持其存活，一个研究小组已经使用这些细胞来研究MDV感染。我们已经使用这些细胞来研究IBDV感染，我们的数据表明，它们将是研究IBDV如何与鸟类免疫系统相互作用的良好模型，而不需要直接感染鸟类。我们相信该模型有可能取代受感染鸟类的使用，并减少此类研究中使用的鸟类总数。本项目的目的是在前期研究的基础上，建立研究IBDV的鸡B细胞模型，并将其扩展到ALV和REV的研究。为此，我们有三个目标：第一、我们将通过全面量化感染后B细胞中基因表达水平的变化，与感染后B细胞中发生的变化相比，确认和验证该模型用于研究IBDV的用途。被感染的鸟第二，我们将扩大模型，包括ALV和REV。第三，我们将使用细胞生产IBDV的股票，目前，病毒股票是通过感染鸟类和从器官中回收病毒，所以如果B细胞可以用于这一目的，它将取代使用更多的感染鸟类。我们还将确定这些细胞是否可以用于筛选疫苗，以确定它们对局部传播和新菌株的有效性，以改善控制。我们不仅能够取代和减少使用受感染的鸡，而且使用B细胞实际上是比使用受感染的鸡更好的方法来回答一些科学上重要的问题，因为只有一个细胞群，而不是许多，并且有可能控制细胞接受的病毒量并知道细胞被感染了多长时间。此外，我们可以在未来应用该模型研究IBDV疫苗如何与细胞相互作用，并研究与多种病毒的共感染，因此该项目不仅具有很高的3Rs影响，而且具有很高的科学和转化影响。