Iniguez Eva, Rodriguez Felipe, Husseneder Claudia, Foil Lane, Maldonado Rosa A
Border Biomedical Research Center, Bioscience Research Building, Department of Biological Sciences, The University of Texas at El Paso, El Paso, TX, United States.
Department of Entomology, Louisiana State University Agricultural Center, Baton Rouge, LA, United States.
Front Cell Infect Microbiol. 2025 May 30;15:1595333. doi: 10.3389/fcimb.2025.1595333. eCollection 2025.
Leishmaniasis and Chagas disease are major human neglected diseases, affecting an estimate of 12 and 6 to 8 million people worldwide, respectively. Current treatments for both diseases are highly toxic for the vertebrate host and lack specificity for the parasites, highlighting the need for the discovery of new therapies against these diseases. In this study, we tested the use of the lytic peptide and a construct that incorporates a ligand to bind the lytic peptide to protozoa membranes and screened them for protozoacidal activity.
We first screened parasite survival of luciferase expressing promastigotes and epimastigotes in the presence of or , and after 12, 48 and 96 h by measuring the parasite luciferase activity. In addition, High-Content Imaging Assay was used to evaluate the proliferation of intracellular amastigotes propagated inside murine macrophages after treatment with or .
The lowest half maximal effective concentration observed after 48 h of incubation with and was lower against promastigotes than epimastigotes. treatment significantly reduced infection rate of macrophages amastigotes compared to the non-treated vehicle control; while treatment with was significant only at higher drug concentrations. Importantly, no significant cytotoxicity was observed when screened against intraperitoneal murine macrophages for either or treatments.
Our results indicate that ligand-lytic peptide complexes are potential targets for therapeutic drugs that can selectively kill both extracellular and intracellular protozoa parasites stages with no significant toxicity to host cells.
利什曼病和恰加斯病是主要的人类被忽视疾病，全球分别估计有1200万人和600万至800万人受其影响。目前这两种疾病的治疗方法对脊椎动物宿主毒性很大，且对寄生虫缺乏特异性，这凸显了发现针对这些疾病的新疗法的必要性。在本研究中，我们测试了溶细胞肽以及一种包含配体的构建体的用途，该配体可使溶细胞肽与原生动物膜结合，并对它们进行杀原虫活性筛选。
我们首先通过测量寄生虫荧光素酶活性，在存在或不存在或的情况下，以及在12、48和96小时后，筛选表达荧光素酶的前鞭毛体和上鞭毛体的寄生虫存活率。此外，使用高内涵成像分析来评估在用或处理后在小鼠巨噬细胞内繁殖的细胞内无鞭毛体的增殖情况。
与上鞭毛体相比，与和孵育48小时后观察到的最低半数最大有效浓度对前鞭毛体更低。与未处理的载体对照相比，处理显著降低了巨噬细胞无鞭毛体的感染率；而仅在较高药物浓度下，处理才有显著效果。重要的是，在用或处理针对腹腔小鼠巨噬细胞进行筛选时，未观察到明显的细胞毒性。
我们的结果表明，配体 - 溶细胞肽复合物是治疗药物的潜在靶点，其可以选择性地杀死细胞外和细胞内原生动物寄生虫阶段，而对宿主细胞无明显毒性。
