Towards the Use of Novel High Density Anopheles-Specific Wolbachia Strains for Anopheles Vector Control.

T1 -基础研究T2 -人类/临床研究在其最新报告中，世界卫生组织估计全球有2.47亿例疟疾病例，619，000例相关死亡。鉴于现有病媒控制方法对减少全世界疟疾负担作出了巨大贡献，最近报告的病媒控制方法效力下降尤其令人关切。亚洲疟疾蚊斯氏按蚊（Anopheles stephensi）是一种适应性很强的城市病媒，它被引入撒哈拉以南非洲地区，给消除疟疾造成了又一个障碍。基于沃尔巴克氏体的干预措施已被证明在针对伊蚊时非常有效，伊蚊是虫媒病毒的主要载体。在最近发表的一项群集随机对照试验中，释放受沃尔巴克氏体感染的蚊子与登革热发病率降低77%和登革热住院率降低86%相关，这不仅证明了基于沃尔巴克氏体的病媒控制策略的可行性，而且证明了沃尔巴克氏体研究的转化性质。鉴于疟疾是绝大多数蚊媒疾病相关发病率和死亡率的原因，因此对开发基于沃尔巴克氏体的疟疾干预措施产生极大兴趣并不奇怪。可行的干预措施的主要障碍是难以产生受沃尔巴克氏体感染的按蚊，只有稳定的交叉感染产生，已知携带沉重的健身成本。直到最近，野生按蚊种群被认为没有沃尔巴克氏体感染，尽管许多蚊子物种和50%的昆虫物种被认为受到感染。最近在两个载体An. demeilloni和An. moucheti的研究已经重振了研究，期望使用按蚊特异性菌株的按蚊Wolbachia交叉感染将证明更成功。拟议的项目存在于发现这些菌株和将其用作干预措施之间的结合点。我将调查按蚊和沃尔巴克氏体之间的独特关系，旨在确定影响按蚊对沃尔巴克氏体易感性的一些因素，同时采取第一步使用这些菌株控制疟疾。目的1识别和减轻限制体外Wolbachia感染的因素1。1建立一个机制模型的框架，以更好地了解限制体外Wolbachia感染成功的因素。1.2在已开发的框架内使用高通量测试，以确定限制Wolbachia感染细胞的因素。1.3比较wAnD的能力，wNo和wAlbB感染伊蚊和按蚊细胞1.4开发用沃尔巴克氏体感染细胞系的优化方案1.5产生用感兴趣的沃尔巴克氏体菌株稳定感染的细胞系。胚胎发育期间沃尔巴克氏体相互作用2.1胚胎发育早期沃尔巴克氏体向性的特征2.2胚胎发育后沃尔巴克氏体向性的特征2.3比较胚胎显微注射Wolbachia后的宿主反应2.4比较不同来源Wolbachia显微注射后蚊子的宿主反应目的3表征自然感染按蚊种群中Wolbachia宿主相互作用3.1表征野生捕获按蚊中Wolbachia的嗜性。moucheti 3.2评估自然感染的An. moucheti母亲

T1 - Basic Research T2 - Human/Clinical ResearchIn its most recent report, the World Health Organisation estimated 247 million cases of malaria, with 619,000 associated deaths worldwide. The recently reported reduction in efficacy of established vector control methods is of particular concern given the vast contribution they have made to the reduction in malaria burden worldwide. The introduction of the Asian Malaria Mosquito, Anopheles stephensi, a highly adaptable urban vector into sub-Saharan Africa presents an additional barrier to malaria elimination. Wolbachia-based interventions have proven highly effective when targeting Aedes mosquitoes, the prominent vectors of arboviruses. In a recently published cluster randomised control trial, the release of Wolbachia infected mosquitoes was associated with an 77% reduction in dengue incidence and an 86% reduction in dengue hospitalisations demonstrating not only the viability of a Wolbachia based vector control strategy but also the translational nature of Wolbachia research. Given that malaria is responsible for the vast majority of mosquito-borne disease related morbidity and mortality, it is unsurprising that there has been great interest in developing a Wolbachia based malaria intervention. The main barrier to a viable intervention has been the difficulty of producing a Wolbachia infected Anopheles mosquito, with the only stable transinfection generated, known to carry a heavy fitness cost. Until recently, wild populations of Anopheles mosquitoes were believed to be free of Wolbachia infections, despite many species of mosquito and 50% of all insect species thought to be infected. The recent discovery of the first two confirmed Anopheles-specific Wolbachia strains, wAnD and wAnM in two vectors An. demeilloni and An. moucheti respectively, has reinvigorated research, with the expectation that Anopheles mosquito Wolbachia transinfections using Anopheles-specific strains will prove more successful. The proposed project exists in the juncture between the discovery of these strains, and their use as an intervention. I will investigate the unique relationship between Anopheles mosquitoes and Wolbachia, aiming to determine some of the factors that influence the susceptibility of Anopheles to Wolbachia while taking the first steps towards the use of these strains for malaria control. Aim 1Identifying and mitigating factors limiting Wolbachia infection in-vitro1.1 Develop the framework for a mechanistic model to better understand factors limiting the success of in-vitro Wolbachia infection. 1.2 Use high throughput testing within the developed framework to identify the factors that limit Wolbachia infection of cells. 1.3 Compare the ability of wAnD, wNo and wAlbB to infect Aedes and Anopheles cells1.4 Develop an optimised protocol for infecting cell lines with Wolbachia 1.5 Generate cell lines stable infected with Wolbachia strains of interest.Aim 2In-vivo analysis of Host-Wolbachia Interactions During Embryonic Development 2.1 Characterising Wolbachia tropism during early embryonic development2.2 Characterising Wolbachia tropism following embryonic microinjection.2.3 Compare the host response upon embryonic microinjection with Wolbachia.2.4 Compare host response in mosquitoes upon microinjection of Wolbachia from different sources.Aim 3Characterise Wolbachia host interaction in naturally infected populations of Anopheles 3.1 Characterise Wolbachia tropism in wild-caught An. moucheti 3.2 Assess host response in offspring of naturally infected An. moucheti mothers