Application of an in vitro cell-based system to investigate the killing of target tissue by drug-specific T-cells

由于它们的性质，药物过敏通常不会被检测到，直到一种药物被大量使用。在这么晚的阶段停药不仅会对人体安全造成风险，而且会给制药公司带来巨大的成本，也会延长患者的时间，直到开发出更安全的药物。发现和开发一种药物的成本估计约为9亿美元，大约需要13年。因此，通过开发评估药物免疫原性的检测方法来减少后期药物停药，将减少将更安全的药物推向市场的成本和时间，从而使制药业和患者受益。药物特异性t细胞已经从过敏患者的血液和组织中分离出来。与特定HLA分子结合的药物激活这些细胞导致细胞因子和细胞溶解分子的分泌。学术导师在药物过敏的化学和细胞基础方面有丰富的经验。为了模拟这种体外反应，我们从1000名健康供体中生成了hla型PBMC库，并设计了研究药物活化naïve t细胞的方法。尽管如此，关于将抗原信号转化为组织损伤所需的免疫和患者因素的基本知识仍然缺失。在此博士学习期间，候选人将探索支持药物过敏的基础科学，并定义与组织损伤发展相关的免疫学参数。该项目将以MRC药物安全科学中心（https://www.liverpool.ac.uk/drug-safety/）为基础。我们是利物浦大学的一个独特的研究中心，致力于了解临床重要药物不良反应的机制。我们将利用来自健康志愿者的表达相关HLA风险等位基因的药物启动t细胞和来自同一供者的诱导多能干细胞肝细胞/角化细胞系。我们将评估药物的免疫原性，但也整合组织和免疫细胞来研究t细胞的活化和靶组织的杀伤。将讨论以下问题：A.哪些药物以限制hla等位基因的方式激活t细胞？B.活化的t细胞的表型和功能是什么？C.药物如何激活t细胞？D.哪些信号通路参与了t细胞介导的组织损伤？只有回答了这些问题，我们才有希望了解药物是如何引起这种医源性疾病的，并开发出预测免疫原性的检测方法。

Due to their nature, drug allergy is usually not detected until a drug is administered to a large population. As well as a risk to human safety, the withdrawal of drugs at such a late stage is of great cost to Pharma, and also prolongs the time for the patient until a safer drug is developed. The cost of discovering and developing a drug is estimated to be about $900 million, and takes approximately 13 years. Thus reducing late-stage drug withdrawals through the development of assays to assess immunogenicity of drugs would reduce the cost and time to bring safer drugs to the market, benefitting the pharmaceutical industry and patients. Drug-specific T-cells have been isolated from blood and tissue of allergic patients. Activation of these cells with the drug bound to specific HLA molecules results in the secretion of cytokines and cytolytic molecules. The academic supervisors have extensive experience characterizing the chemical and cellular basis of drug allergy. To model such reactions ex vivo we have generated an HLA-typed bank of PBMC from 1000 healthy donors and designed methods to study the activation naïve T-cells with drugs. Despite this, essential knowledge of the immune and patient factors required to convert the antigenic signal into tissue injury is missing. During this PhD studentship, the candidate will explore the fundamental science underpinning drug allergy and define the immunological parameters associated the development of tissue injury. The project will be based in the MRC Centre for Drug Safety Science (https://www.liverpool.ac.uk/drug-safety/). We are a unique research centre at The University of Liverpool dedicated to understanding the mechanisms of clinically-important adverse drug reactions. We will utilize drug-primed T-cells from healthy volunteers expressing relevant HLA risk alleles and induced pluripotent stem cell hepatocytes/keratinocytes-lines derived from the same donors. We will assess the immunogenicity of drugs, but also integrate the tissue and immune cells to investigate T-cell activation and the killing of target tissue. The following questions will be addressed: A. Which drugs activate T-cells in an HLA-allele-restricted manner? B. What is the phenotype and function of the activated T-cells? C. How do drugs activate T-cells? and D. Which signalling pathways are involved in T-cell-mediated tissue injury? Only by answering these questions can we hope to understand how drugs cause this form of iatrogenic disease and develop assays to predict immunogenicity.