Proteomic profile of plasma-derived extracellular vesicles from Colombian pregnant women with Plasmodium-soil transmitted helminths coinfection

Background: Knowledge about extracellular vesicles (EV) and their molecular cargo in gestational parasitic infections, particularly by Plasmodium and soil-transmitted helminths (STH), is almost non-existent. Objective: To perform isolation and molecular characterization of plasma-derived EVs from Colombian pregnant women and compare quantity, size, concentration and protein cargo of those EVs according to the infectious status. Methodology: Five study groups were formed: 1), Pregnant women with Plasmodium infection. 2), Pregnant women with STH infection. 3), Pregnant women with coinfection Plasmodium and STH. 4), Pregnant women without infection with Plasmodium nor STH. 5), Non-pregnant women without infection with Plasmodium nor STH. Plasma-derived EVs were isolated by size exclusion chromatography (SEC) and fractions containing EVs identified by a bead-based flow cytometric assay for tetraspanin CD9; the size and concentration of EVs were quantified by nanoparticle tracking analysis, and proteins associated with EVs were identified by liquid chromatography-mass spectrometry (LC-MS) in a pool of samples per study group.

研究背景：目前关于妊娠寄生虫感染（尤其是疟原虫与土源性蠕虫（soil-transmitted helminths, STH）感染）相关的细胞外囊泡（extracellular vesicles, EV）及其分子载荷的研究资料几乎空白。研究目的：对哥伦比亚孕妇血浆来源的细胞外囊泡进行分离与分子表征，并依据感染状态对比这些细胞外囊泡的数量、粒径、浓度及蛋白载荷。研究方法：本研究共设置5个组别：1）疟原虫感染孕妇组；2）土源性蠕虫感染孕妇组；3）疟原虫与土源性蠕虫共感染孕妇组；4）未感染疟原虫及土源性蠕虫的健康孕妇组；5）未感染疟原虫及土源性蠕虫的非妊娠女性组。采用尺寸排阻色谱法（size exclusion chromatography, SEC）分离血浆来源的细胞外囊泡，通过针对四跨膜蛋白CD9的磁珠流式细胞术鉴定含有细胞外囊泡的组分；采用纳米颗粒追踪分析量化细胞外囊泡的粒径与浓度，并通过液相色谱-质谱联用法（liquid chromatography-mass spectrometry, LC-MS）对每个研究组的混合样本进行细胞外囊泡相关蛋白的鉴定。