Molecular Definition Of Filarial And Related Nonfilarial Genes And Proteins

完成了罗阿罗阿的基因组后，W。bancrofti和（最近的）O.volvulus，我们利用基因组数据作为背景进行大量的蛋白质组学和转录组学研究。 我们已经完成了一个大规模的蛋白质组和转录组的特点，几乎所有的主要哺乳动物阶段的O。肠扭转，导致超过85-90%的预测产品的鉴定从O。肠扭转基因组/推定蛋白质组。分析还获得了沃尔巴克氏体的大部分蛋白质组，沃尔巴克氏体是O.肠扭转 寄生虫性别和阶段特异性蛋白表达确定了这些途径与寄生虫分化。 为了更好地理解强调蚊子衍生的感染期幼虫（L3）到哺乳动物适应的L3和蜕皮后到L4之间的过渡以及适应人类宿主的最初一周的发育程序，我们适应了允许L3发育和随后蜕皮到L4的体外系统。 通过这个9天的过程多次使用微阵列和蛋白质组学评估，我们不仅鉴定了对L3/L4转变至关重要的那些基因/途径，而且我们还通过药理学抑制（半胱氨酸蛋白酶抑制）和RNAi（关键CPL）证明了半胱氨酸蛋白酶在哺乳动物适应性L3至L4的早期发育中所起的关键作用。 我们最近对确定的丝虫感染患者的血清、Loa loa微丝蚴的排泄/分泌（E/S）产物、O. vovlulus蠕虫和适当的对照以鉴定活性感染的寄生虫来源的生物标志物。 这导致了分子靶点的鉴定，这些分子靶点已被用于配置用于快速检测O.肠扭转和Loa loa。 我们通过评估阿苯达唑（ALB）治疗前3周和3个月后从5个肯尼亚农村村庄的居民收集的粪便，研究了土壤传播的蠕虫在改变肠道微生物组中所起的作用。有趣的是，与年龄匹配的对照组相比，蛔虫和美洲钩虫感染的存在都没有显著改变微生物群的总体多样性，尽管在ALB治疗和土壤传播蠕虫（STH）清除后，梭菌目构成的微生物群比例显著增加（肠杆菌目构成的比例减少）。 处理后丰富度也显著降低。我们的数据表明，通过驱虫清除STH改变了肠道微生物群。 我们已经产生了一个296兆碱基（Mb）的参考质量的基因组组成的17902蛋白编码基因来自一个单一的，有代表性的蛔虫从60个人类宿主在肯尼亚的村庄，养猪业是罕见的。值得注意的是，大多数人分离株（63/68）的线粒体基因组与猪蛔虫的距离比与A.蚯蚓。比较基因组分析确定了超过1100万个核编码的SNP，但只有两种不同的遗传类型在分析的基因组中重组。核基因组具有广泛的杂合性，所有样品都存在与A。suum-like或A.支持高度杂交蛔虫物种遗传复合体的类蛔虫遗传模式。由于这些混合蠕虫的人传人似乎不存在障碍，因此有必要采取单一健康方法来控制人类蛔虫病的传播。 microRNA为O.已经在盘尾丝虫病患者的血清中鉴定了肠扭转，并且已经鉴定了基于miRNA多态性的区域差异。

Having completed the genomes of Loa loa, W. bancrofti, and (most recently) O.volvulus, we have utilized the genomic data as the backdrop for performing a large number of proteomic and transcriptomic studies. We have completed a large-scale proteomic and transcriptomic characterization of almost all the major mammalian stages of O. volvulus, resulting in the identification of more than 85-90% of the products predicted from the O. volvulus genome/putative proteome. The analysis also yielded much of the proteome of Wolbachia, the obligate endosymbiont of O. volvulus. Parasite sex- and stage-specific protein expression identified those pathways related to parasite differentiation. To understand better the developmental programs that underscore the transition between the mosquito-derived infective stage larvae (L3) to mammalian adapted L3s and to L4s following a molt, and the initial week of adaptation to the human host, we adapted an in vitro system that allowed for L3 development and subsequent molting to the L4. Using microarray and proteomic assessments at multiple times through this 9 day process we have not only identified those genes/pathways that are critical for the L3/L4 transition but we have also demonstrated by both pharmacologic inhibition (cysteine protease inhibition) and RNAi (of the critical CPLs) the critical role played by cysteine proteases in the early development of mammalian adapted L3s to L4s. We have recently performed shotgun mass spectroscopy on both human sera of patients with defined filarial infections, excretory/secretory (E/S) products of Loa loa microfilariae, all stages of the O. vovlulus worm, and appropriate controls to identify parasite derived biomarkers of active infection. This has led to identification of molecular targets that have been used d to configure quantitative immunoassays for the rapid detection of active infection for O. volvulus and Loa loa. We examined the role played by soil transmitted helminths in altering the intestinal microbiome by assessing stools collected from residents of 5 rural Kenyan villages prior to and 3 weeks and 3 months following albendazole (ALB) therapy. Interestingly,, the presence of neither Ascaris lumbricoides nor Necator americanus infection significantly altered the overall diversity of the microbiota in comparison with age-matched controls, although Following ALB therapy and clearance of soil-transmitted helminths (STH), there were significant increases in the proportion of the microbiota made up by Clostridiales ( and reductions in the proportion made up by Enterobacteriales. There was also a significant posttreatment decrease in richness, Our data suggest that clearance of STH through deworming alters the gut microbiota. We have generated a 296 megabase (Mb) reference quality genome comprised of 17902 protein-coding genes derived from a single, representative Ascaris worm collected from 60 human hosts in Kenyan villages where pig husbandry is rare. Notably, the majority of human isolates (63/68) possessed mitochondrial genomes that clustered closer to the pig parasite Ascaris suum than to A. lumbricoides. Comparative phylogenomic analyses identified over 11 million nuclear-encoded SNPs but just two distinct genetic types that had recombined across the genomes analysed. The nuclear genomes had extensive heterozygosity and all samples existed as genetic mosaics with either A. suum-like or A. lumbricoides-like inheritance patterns supporting a highly interbred Ascaris species genetic complex. As no barriers appear to exist for anthroponotic transmission of these hybrid worms, a one-health approach to control the spread of human ascariasis will be necessary. MicroRNA for O. volvulus have been identified in the sera of patients with onchocerciasis and regional differences based on miRNA polymorphisms have been identified.