Structure-function relationships of Toll-like receptors, key mediators of antiviral innate immunity, in Human Papillomavirus 16 infection and cervical cancer

toll样受体（TLRs）是一类细胞表面受体，在对细菌、病毒和其他微生物入侵的应答中，参与先天激活和随后的适应性免疫。TLRs检测多种保守分子结构，如细菌脂多糖（TLR4）、脂蛋白（TLR2）；细菌DNA (TLR9)；病毒RNA (TLR3和7)，病毒蛋白（TLRs 2和4），但配体识别，受体激活和细胞内信号传导的分子原理在很大程度上仍然未知。人类乳头瘤病毒（HPV） 16是宫颈癌的主要病因，也是全球女性癌症死亡的第二大原因。有证据表明，先天免疫反应的组成部分对控制病毒感染至关重要。tlr是HPV免疫识别的主要候选者之一。因此，tlr在对抗hpv16感染和预防宫颈癌方面具有巨大的潜力。本项目试图从结构-功能的角度解决TLR生物学的关键方面。利用可用的（来自蛋白质晶体结构）和可推导的（通过比较建模）结构信息，采用基于诱变的功能分析来识别对信号传导重要的氨基酸。随后，诱变效应将使用生化和生物物理方法进行更详细的研究。此外，tlr在HPV16免疫中的作用将被研究。TLR单核苷酸多态性（SNP）与持续性HPV16感染/宫颈癌之间的相关性将通过将SNP信息引入已建立的基于诱变的功能筛选中来分析。此外，TLR的表达和受体刺激对病毒感染进展的影响将在HPV感染模型中进行研究。

Toll-like receptors (TLRs) are a family of cell surface receptors required for innate activation and subsequent adaptive immunity in response to invasion by bacteria, viruses, and other microbes. TLRs detect a multitude of conserved molecular structures such as bacterial lipopolysaccharide (TLR4), lipoproteins (TLR2); bacterial DNA (TLR9); viral RNA (TLR3 and 7), viral proteins (TLRs 2 and 4) but the molecular principles of ligand recognition, receptor activation and intracellular signalling remain largely unknown. Human Papillomavirus (HPV) 16 is the primary etiological agent for cervical cancer and the second biggest cause of female cancer mortality worldwide. There is evidence that components of the innate immune response are crucial in controlling viral infection. TLRs are among the primary candidates for HPV immune recognition. Consequently, TLRs have an enormous potential in the fight against HPV16-infection and the prevention of cervical cancer. This project attempts to address key aspects of TLR biology from a structure-function perspective. Using available (from protein crystal structures) and deducible (by comparative modelling) structural information, a functional assay based on mutagenesis is employed to identify amino acids of importance for signalling. Subsequently, the effects of mutagenesis will be investigated in more detail using biochemical and biophysical methods. Additionally, the role of TLRs in HPV16 immunity will be investigated. Correlations between TLR single nucleotide polymorphisms (SNPs) and persistent HPV16 infection/cervical cancer will be analyzed by introducing SNP information into the established mutagenesis-based functional screen. Furthermore, TLR expression and the influence of receptor stimulation on the progression of viral infection will be studied in an HPV model of infection.