Department of Chemistry and Biochemistry, University of Oklahoma, Norman; Laboratories of Molecular Anthropology and Microbiome Research, University of Oklahoma, Norman.
Laboratories of Molecular Anthropology and Microbiome Research, University of Oklahoma, Norman; Department of Anthropology, University of Oklahoma, Norman.
J Vis Exp. 2022 Jan 21(179). doi: 10.3791/63255.
Pathogen tropism and disease tropism refer to the tissue locations selectively colonized or damaged by pathogens, leading to localized disease symptoms. Human-infective trypanosomatid parasites include Trypanosoma cruzi, the causative agent of Chagas disease; Trypanosoma brucei, the causative agent of sleeping sickness; and Leishmania species, causative agents of leishmaniasis. Jointly, they affect 20 million people across the globe. These parasites show specific tropism: heart, esophagus, colon for T. cruzi, adipose tissue, pancreas, skin, circulatory system and central nervous system for T. brucei, skin for dermotropic Leishmania strains, and liver, spleen, and bone marrow for viscerotropic Leishmania strains. A spatial perspective is therefore essential to understand trypanosomatid disease pathogenesis. Chemical cartography generates 3D visualizations of small molecule abundance generated via liquid chromatography-mass spectrometry, in comparison to microbiological and immunological parameters. This protocol demonstrates how chemical cartography can be applied to study pathogenic processes during trypanosomatid infection, beginning from systematic tissue sampling and metabolite extraction, followed by liquid chromatography-tandem mass spectrometry data acquisition, and concluding with the generation of 3D maps of metabolite distribution. This method can be used for multiple research questions, such as nutrient requirements for tissue colonization by T. cruzi, T. brucei, or Leishmania, immunometabolism at sites of infection, and the relationship between local tissue metabolic perturbation and clinical disease symptoms, leading to comprehensive insight into trypanosomatid disease pathogenesis.
病原体嗜性和疾病嗜性是指病原体选择性定植或破坏的组织部位，导致局部疾病症状。能感染人类的锥体虫包括引起恰加斯病的克氏锥虫、引起昏睡病的布氏锥虫和引起利什曼病的利什曼原虫。这些寄生虫共同影响着全球 2000 万人。它们具有特定的嗜性：T. cruzi 感染心脏、食道和结肠；T. brucei 感染脂肪组织、胰腺、皮肤、循环系统和中枢神经系统；皮肤利什曼原虫感染皮肤；内脏利什曼原虫感染肝脏、脾脏和骨髓。因此，从空间角度理解锥体虫病发病机制至关重要。化学绘图通过液相色谱-质谱法生成小分子丰度的 3D 可视化，与微生物学和免疫学参数进行比较。本方案演示了如何将化学绘图应用于研究锥体虫感染过程中的致病过程，从系统组织采样和代谢物提取开始，接着进行液相色谱-串联质谱数据采集，最后生成代谢物分布的 3D 图谱。这种方法可用于多种研究问题，如 T. cruzi、T. brucei 或利什曼原虫对组织定植的营养需求、感染部位的免疫代谢以及局部组织代谢紊乱与临床疾病症状之间的关系，从而全面了解锥体虫病发病机制。
