de Rezende Eloiza, Kawahara Rebeca, Peña Mauricio S, Palmisano Giuseppe, Stolf Beatriz S
Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
PLoS Negl Trop Dis. 2017 Nov 27;11(11):e0006090. doi: 10.1371/journal.pntd.0006090. eCollection 2017 Nov.
Leishmaniasis is an antropozoonosis caused by Leishmania parasites that affects around 12 million people in 98 different countries. The disease has different clinical forms, which depend mainly on the parasite genetics and on the immunologic status of the host. The promastigote form of the parasite is transmitted by an infected female phlebotomine sand fly, is internalized by phagocytic cells, mainly macrophages, and converts into amastigotes which replicate inside these cells. Macrophages are important cells of the immune system, capable of efficiently killing intracellular pathogens. However, Leishmania can evade these mechanisms due to expression of virulence factors. Different strains of the same Leishmania species may have different infectivity and metastatic phenotypes in vivo, and we have previously shown that analysis of amastigote proteome can give important information on parasite infectivity. Differential abundance of virulence factors probably accounts for the higher virulence of PH8 strain parasites shown in this work. In order to test this hypothesis, we have quantitatively compared the proteomes of PH8 and LV79 lesion-derived amastigotes using a label-free proteomic approach.
METHODOLOGY/PRINCIPAL FINDINGS: In the present work, we have compared lesion development by L. (L.) amazonensis PH8 and LV79 strains in mice, showing that they have different virulence in vivo. Viability and numbers of lesion-derived amastigotes were accordingly significantly different. Proteome profiles can discriminate parasites from the two strains and several proteins were differentially expressed.
CONCLUSIONS/SIGNIFICANCE: This work shows that PH8 strain is more virulent in mice, and that lesion-derived parasites from this strain are more viable and more infective in vitro. Amastigote proteome comparison identified GP63 as highly expressed in PH8 strain, and Superoxide Dismutase, Tryparedoxin Peroxidase and Heat Shock Protein 70 as more abundant in LV79 strain. The expression profile of all proteins and of the differential ones precisely classified PH8 and LV79 samples, indicating that the two strains have proteins with different abundances and that proteome profiles correlate with their phenotypes.
利什曼病是一种由利什曼原虫寄生虫引起的人兽共患病,在98个不同国家影响着约1200万人。该疾病有不同的临床形式,主要取决于寄生虫的遗传学和宿主的免疫状态。寄生虫的前鞭毛体形式由受感染的雌性白蛉传播,被吞噬细胞(主要是巨噬细胞)内化,并转化为无鞭毛体,在这些细胞内复制。巨噬细胞是免疫系统的重要细胞,能够有效杀死细胞内病原体。然而,利什曼原虫可因毒力因子的表达而逃避这些机制。同一利什曼原虫物种的不同菌株在体内可能具有不同的感染性和转移表型,我们之前已经表明,对无鞭毛体蛋白质组的分析可以提供有关寄生虫感染性的重要信息。毒力因子丰度的差异可能是本研究中PH8菌株寄生虫毒力较高的原因。为了验证这一假设,我们使用无标记蛋白质组学方法对PH8和LV79病变来源的无鞭毛体的蛋白质组进行了定量比较。
方法/主要发现:在本研究中,我们比较了亚马逊利什曼原虫PH8和LV79菌株在小鼠体内的病变发展情况,表明它们在体内具有不同的毒力。病变来源的无鞭毛体的活力和数量因此存在显著差异。蛋白质组图谱可以区分这两种菌株的寄生虫,并且有几种蛋白质存在差异表达。
结论/意义:这项研究表明,PH8菌株在小鼠中更具毒力,并且该菌株病变来源的寄生虫在体外更具活力和感染性。无鞭毛体蛋白质组比较确定GP63在PH8菌株中高表达,超氧化物歧化酶、锥虫过氧化物酶和热休克蛋白70在LV79菌株中更为丰富。所有蛋白质和差异蛋白质的表达谱精确地对PH8和LV79样本进行了分类,表明这两种菌株具有不同丰度的蛋白质,并且蛋白质组图谱与其表型相关。