Department of Veterinary Medicine and Surgery, University of Missouri, Columbia, Missouri, USA.
Skin and Stem Cell Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Parasite Immunol. 2024 Oct;46(10):e13068. doi: 10.1111/pim.13068.
Leishmaniasis is a parasitic disease spread by the bite of an infected sandfly and caused by protozoan parasites of the genus Leishmania. Currently, there is no vaccine available for leishmaniasis in humans, and the existing chemotherapy methods face various clinical challenges. The majority of drugs are limited to a few toxic compounds, with some parasite strains developing resistance. Therefore, the discovery and development of a new anti-leishmanial compound is crucial. One promising strategy involves the use of nanoparticle delivery systems to accelerate the effectiveness of existing treatments. In this study, Amphotericin B (AmB) was incorporated into functionalized carbon nanotube (f-CNT) and evaluated for its efficacy against Leishmania major in vitro and in a BALB/c mice model. The increase in footpad thickness was measured, and real-time PCR was used to quantify the parasite load post-infection. Levels of nitric oxide and cytokines IL-4 and IFN-γ were also determined. We found that f-CNT-AmB significantly reduced the levels of promastigotes and amastigotes of the Leishmania parasite. The nanoparticle showed strong anti-leishmanial activity with an IC of 0.00494 ± 0.00095 mg/mL for promastigotes and EC of 0.00294 ± 0.00065 mg/mL for amastigotes at 72 h post-infection, without causing harm to mice macrophages. Treatment of infected BALB/c mice with f-CNT-AmB resulted in a significant decrease in cutaneous leishmania (CL) lesion size in the foot pad, as well as reduced Leishmania burden in both lymph nodes and spleen. The levels of nitric oxide and IFN-γ significantly increased in the f-CNT-AmB treated groups. Also, our results showed that the level of IL-4 significantly decreased after f-CNT-AmB treatment in comparison to other groups. In conclusion, our results demonstrate that AmB loaded into f-CNT is significantly more effective than AmB alone in inhibiting parasite propagation and promoting a shift towards a Th1 response.
利什曼病是一种寄生虫病，通过受感染的沙蝇叮咬传播，由利什曼原虫属的原生动物寄生虫引起。目前，人类尚无针对利什曼病的疫苗，现有的化疗方法面临各种临床挑战。大多数药物仅限于少数几种有毒化合物，一些寄生虫株已产生耐药性。因此，发现和开发新的抗利什曼化合物至关重要。一种有前途的策略是利用纳米颗粒递送系统来加速现有治疗方法的效果。在这项研究中，两性霉素 B（AmB）被包裹在功能化碳纳米管（f-CNT）中，并评估其在体外和 BALB/c 小鼠模型中对抗利什曼原虫的功效。通过测量足垫厚度来评估，并用实时 PCR 来定量感染后寄生虫负荷。还测定了一氧化氮和细胞因子 IL-4 和 IFN-γ 的水平。我们发现，f-CNT-AmB 可显著降低利什曼原虫的前鞭毛体和无鞭毛体寄生虫的水平。纳米颗粒表现出强大的抗利什曼活性，其对前鞭毛体的 IC 为 0.00494±0.00095mg/mL，对无鞭毛体的 EC 为 0.00294±0.00065mg/mL，在感染后 72 小时，对小鼠巨噬细胞没有造成伤害。用 f-CNT-AmB 治疗感染的 BALB/c 小鼠可显著减小足部皮肤利什曼病变的大小，并降低淋巴结和脾脏中的利什曼虫负荷。f-CNT-AmB 处理组的一氧化氮和 IFN-γ水平显著升高。此外，与其他组相比，f-CNT-AmB 处理后，IL-4 的水平显著降低。总之，我们的结果表明，负载 AmB 的 f-CNT 比单独使用 AmB 更能有效地抑制寄生虫繁殖，并促进向 Th1 反应转变。
