Improving TB vaccine design by natural selection of intracellularly processed epitopes presented in primary tissues of murine model for protective imm

通过自然选择鼠模型原代组织中存在的细胞内加工表位来改进结核疫苗设计，以实现保护性免疫

• 批准号: 1923675
• 金额: --
• 依托单位: St George's, University of London
• 依托单位国家: 英国
• 项目类别: Studentship
• 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=studentship-1923675
• 关键词: Improving; TB; vaccine; design; natural

SUMMARYOne of the main hallmarks of Mycobacterium tuberculosis (MTB) pathogenesis is its ability to manipulate the timing of the immune response to its own advantage by changing expression of its antigens to avoid early recognition and destruction. MTB has capacity for infection of a wide range of tissues [Fanning A, 1999] with extrapulmonary TB representing approximately 15% of newly reported cases and 20-30% of relapsed cases [Global Tuberculosis Report 2017, WHO] and while macrophages constitute its main primary host, other cell types, such as professional phagocytes (i.e. dendritic cells) and non-professional antigen presenting cells (APCs) i.e. epithelial cells are submissive to the pathogen [Mvubu NE 2016, Harrif MJ, 2014, Lerner TR 2016] and can process and present MTB antigens [Flyer DC 2002, Penn BH 2018]. Cross-presentation of foreign antigens by host is a critical part of the process and in MTB infection this predominantly involves MHC-I receptors associated with short peptides derived via the intracellular proteasome-focused processing pathway [Adiko AC, 2015; Chen H, 2016; Ivanyi J, 2014]. Understanding and being able to influence this system is a fundamental part of next generation vaccine design. HYPOTHESISThe leading project hypothesis is that stimulation of Mtb antigens cross-presentation pathways may translate in identification of new protective antigens for next generation of vaccines. AIMSObjective 1. To examine BCG and MTB presentation upon in vitro infection in various mouse organs grouped as follows: Primary and secondary lymphoid organs Urinary system Respiratory system Cardiac system Differentiated bone marrow cells Objective 2. To ascertain antigenic changes in BCG and MTB presentation upon cross-presentation pathways stimulation METHODS AND TECHNIQUES- organ collection for in vitro infection assays and tissue culture - extraction and purification of immunogenic complexes - antigen(s) identification via mass spectrometry (collaborative work) FUTURE PROSPECTSAny new identified antigens / methods enhancing their presentation may be included into the current TB vaccination strategies to improve their efficacy REFERENCES1. https://www.who.int/tb/publications/global_report/gtbr2017_main_text.pdf 2. Adiko AC et al. 'Intracellular transport routes for MHC I and their relevance for antigen crosspresentation' Front Immunol. 2015 Jul 2;6:335. doi: 10.3389/fimmu.2015.00335. eCollection 2015. Page 1 of 2 3. Fanning A. Tuberculosis: 6. Extrapulmonary disease; CMAJ. 1999 Jun 1;160(11):1597-603; PMID: 10374005 4. Flyer DC et al. Identification by mass spectrometry of CD8(+)-T-cell Mycobacterium tuberculosis epitopes within the Rv0341 gene product' Infect Immun. 2002 Jun;70(6):292632. 5. Harriff MJ et al. 'Human lung epithelial cells contain Mycobacterium tuberculosis in a late endosomal vacuole and are efficiently recognized by CD8 T cells' PLoS One. 2014 May 14;9(5):e97515. doi: 10.1371/journal.pone.0097515. eCollection 2014. 6. Ivanyj J 'Function and potentials of M.tuberculosis epitopes' Front Immunol. 2014 Mar 24;5:107. doi: 10.3389/fimmu.2014.00107. eCollection 2014. 7. Keller Ch et al. 'Genetically determined susceptibility to tuberculosis in mice causally involves accelerated and enhanced recruitment of granulocytes' Infect Immun. 2006 Jul;74(7):4295-309. 8. Lerner TR et al. Lymphatic endothelial cells are a replicative niche for Mycobacterium tuberculosis J Clin Invest. 2016 Mar 1;126(3):1093-108. doi: 10.1172/JCI83379. Epub 2016 Feb 22. 9. Mvubu NE et al. 'Mycobacterium tuberculosis strains exhibit differential and strain-specific molecular signatures in pulmonary epithelial cells' Dev Comp Immunol. 2016 Dec;65:321329. doi: 10.1016/j.dci.2016.07.022. Epub 2016 Aug 3. 10. Penn BH et al. 'An Mtb-Human Protein-Protein Interaction Map Identifies a Switch between Host Antiviral and Antibacterial Responses' Mol Cell. 2018 Aug 16;71(4):637-648.e5.

结论结核分枝杆菌(MTB)致病机制的主要特征之一是它能够通过改变其抗原的表达来操纵免疫反应的时机，以避免早期识别和破坏。结核分枝杆菌具有感染多种组织的能力，肺外结核约占新报告病例的15%和复发病例的20%-30%[2017年全球结核病报告，世卫组织]，虽然巨噬细胞是其主要宿主，但其他类型的细胞，如专业性吞噬细胞(即树突状细胞)和非专业性抗原提呈细胞(APC)，即上皮细胞服从病原体[Mvubu NE 2016，Harrif MJ，2014，Lerner Tr2016]，并能处理和递送MTB抗原[Flyer DC 2002，Penn BH 2018]。宿主对外来抗原的交叉递呈是这一过程的关键部分，在结核分枝杆菌感染中，这主要涉及与通过细胞内蛋白酶体集中处理途径获得的短肽相关的MHC-I受体[Adiko AC，2015；Chen H，2016；依万尼J，2014]。理解并能够影响这一系统是下一代疫苗设计的基本部分。HYPOTHISIS主要的项目假说是，Mtb抗原交叉递呈途径的刺激可能会转化为为下一代疫苗识别新的保护性抗原。目的1.研究卡介苗和结核分枝杆菌在小鼠原发和次级淋巴器官、泌尿系统、呼吸系统、心脏系统分化的骨髓细胞体外感染中的呈递情况。目的2.探讨卡介苗和结核杆菌呈递在交叉呈递途径、刺激方法和技术上的抗原变化--体外感染检测和组织培养的器官采集-免疫原复合体的提取和纯化-质谱学鉴定(S)抗原(协作性工作)未来PROSPECTS任何新识别的抗原/方法可能被纳入当前的结核病疫苗接种策略中，以提高其效力。Https://www.who.int/tb/publications/global_report/gtbr2017_main_text.pdf 2.Adiko AC等人。MHC I的细胞内转运途径及其与抗原交叉压强的相关性。2015年7月2日；6：335。DOI：10.3389/fimmu.2015.00335.ECollection 2015年。第1页，共2页3.肺结核：6.肺外疾病；CMAJ.1999年6月1日；第160(11)：1597-603；PMID：10374005。Rv0341基因产物感染免疫中CD8(+)-T细胞结核分枝杆菌表位的质谱学鉴定2002年6月；70(6)：292632。5.Harriff MJ等人。人肺上皮细胞在晚期的内体空泡中含有结核分枝杆菌，并能被CD8T细胞的PLoS One有效识别。2014年5月14日；9(5)：e97515。DOI：10.1371/Joural.pone.0097515.ECollection 2014年。6.结核分枝杆菌前免疫表位的伊万尼J‘功能及其潜能2014年3月24日；5：107。DOI：10.3389/fimmu.2014.00107.ECollection 2014年。7.Keller CH等人。基因决定的小鼠对结核病的易感性与感染免疫的粒细胞的加速和增强募集有关。2006年7月；74(7)：4295-309。8.Lerner tr等人。淋巴管内皮细胞是结核分枝杆菌J Clin Invest的复制利基。2016年3月1日；126(3)：1093-108。DOI：10.1172/JCI83379。EPub 2016 2月22日9.Mvubu Ne等人。结核分枝杆菌菌株在肺上皮细胞的Dev comp免疫中表现出不同的和菌株特有的分子特征。2016年12月；65：321329。DOI：10.1016/j.dci.2016.07.022.EPub 2016年8月3日。10.Penn BH等人。结核分枝杆菌-人类蛋白质相互作用图确定了宿主抗病毒和抗菌反应之间的切换摩尔细胞。2018年8月16日；71(4)：637-648.e5。