Martínez-Orellana Pamela, Baxarias Marta, Good Liam, Solano-Gallego Laia
Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain.
Department of Pathobiology and Population Sciences, The Royal Veterinary College, London NW1 0NH, UK.
Vet Sci. 2020 Nov 13;7(4):179. doi: 10.3390/vetsci7040179.
Dogs are the main reservoir for manifesting from a subclinical to a fatal disease. Limited treatments are available, although new antiparasitics and immunomodulators are pursued. Polyhexamethylene biguanide (PHMB) has a broad antimicrobial spectrum, including antiparasitic activity. Here, we evaluated the potential for Toll-like receptor agonists (TLRa) and PHMB alone, and as polyplex nanoparticles containing PHMB and TLR4 or TLR9 agonists, to selectively kill . Susceptibility of promastigotes to PHMB, miltefosine, and allopurinol was performed, and the half-maximum inhibitory concentrations (IC) were determined. Then, DH-82 cells were infected and treated with PHMB alone or combined with TLR4a (MPLA-SM) or TLR9a (CpG ODNs) and allopurinol alone. The IC values of promastigotes were PHMB (1.495 µM), miltefosine (9.455 µM), and allopurinol (0.124 µM). After infection, treated DH-82 cells displayed a lower percentage ( 0.0316), intensity ( 0.0002), and index of infection ( 0.0022) when compared to non-treated cells. PHMB induced lower percentage of infection alone ( 0.043), in combination with TLR9a ( 0.043), and with TLR4a ( 0.0213). Supernatants were collected and used to measure TNF-α and IL-6 levels. Increased TNF-α was observed after PHMB TLR4a, relative to uninfected and infected untreated macrophages ( 0.043). PHMB combined with TLR4a shows promise as a potential anti- drug combination, as well as inducer of proinflammatory response, as demonstrated by decreased infection and increased TNF-α production.
狗是从亚临床疾病发展为致命疾病的主要宿主。尽管正在研发新的抗寄生虫药和免疫调节剂，但可用的治疗方法有限。聚六亚甲基双胍（PHMB）具有广泛的抗菌谱，包括抗寄生虫活性。在此，我们评估了Toll样受体激动剂（TLRa）和单独的PHMB，以及作为包含PHMB和TLR4或TLR9激动剂的复合纳米颗粒，选择性杀死[具体寄生虫名称未给出]的潜力。进行了前鞭毛体对PHMB、米替福新和别嘌呤醇的敏感性实验，并测定了半数最大抑制浓度（IC）。然后，用单独的PHMB或与TLR4a（MPLA-SM）或TLR9a（CpG ODNs）以及单独的别嘌呤醇联合处理感染DH-82细胞。前鞭毛体的IC值分别为：PHMB（1.495 µM）、米替福新（9.455 µM）和别嘌呤醇（0.124 µM）。感染后，与未处理的细胞相比，经处理的DH-82细胞显示出更低的感染百分比（0.0316）、感染强度（0.0002）和感染指数（0.0022）。单独使用PHMB（0.043）、与TLR9a联合使用（0.043）以及与TLR4a联合使用（0.0213）时，感染百分比均较低。收集上清液并用于测量TNF-α和IL-6水平。与未感染和感染未处理的巨噬细胞相比，PHMB与TLR4a联合处理后观察到TNF-α升高（0.043）。PHMB与TLR4a联合显示出作为潜在抗[寄生虫名称未给出]药物组合以及促炎反应诱导剂的前景，这通过感染减少和TNF-α产生增加得以证明。
