Center for Doctoral Studies on Health Sciences (Immunopathology), Faculty of Medicine and Pharmacy, Hassan II University of Casablanca (UH2C), Casablanca, Morocco.
Research team on Immunopathology of Infectious And Systemic Diseases, Laboratory of Cellular and Molecular Pathology, Faculty of Medicine and Pharmacy, UH2C, Casablanca, Morocco.
Parasit Vectors. 2017 Oct 23;10(1):506. doi: 10.1186/s13071-017-2401-4.
BACKGROUND: Cutaneous leishmaniasis (CL) is a vector-borne parasitic disease caused by protozoa of the genus Leishmania. In Morocco, CL is a public health problem mainly caused by Leishmania major and Leishmania tropica, which are responsible for zoonotic and anthroponotic CL, respectively. Macrophages are the primary cells infected by Leishmania parasites and their capacity to produce nitric oxide (NO) is of critical importance for parasite elimination. To our knowledge, the role of NO on autochthonous infections has never been investigated before. In this study, we evaluated in vitro the capacity of autochthonous primary dermotropic strains of L. major and L. tropica to modulate NO production by J774-macrophages and determine the sensitivity of both species to exogenous NO. METHODS: The infectivity of the J774 cell line was analyzed by optical microscopy. NO production by macrophages was measured by the Griess method. The sensitivity to NO by the two strains was assessed by the MTT assay using NO donors. RESULTS: Our results show that the percentage of infected macrophages and the average number of parasites per macrophage were similar for L. major and L. tropica strains. While L. tropica significantly inhibited NO production induced by LPS and IFN-γ stimulation in J774 macrophages, L. major did not affect it. However, soluble Leishmania antigens (SLAs) from both autochthonous primary strains significantly inhibited the production of NO by J774-macrophages in a dose-dependent manner. Finally, our results demonstrated that promastigotes and amastigotes from both strains are sensitive to SNAP NO donor in a dose-dependent manner, although L. tropica demonstrated an increased sensitivity. CONCLUSIONS: Our results suggest a differential ability of L. major and L. tropica strains to modulate the capacity of macrophages to produce NO. The increased ability of L. tropica to inhibit NO production by macrophages might come as a necessity due to its higher sensitivity to NO donor. Our results provide one explanation for the tendency of L. tropica to cause chronic lesions and may contribute to the different physiopathology of CL in Morocco.
背景：皮肤利什曼病（CL）是一种由原生动物利什曼原虫引起的虫媒寄生虫病。在摩洛哥，CL 是一个公共卫生问题，主要由利什曼原虫引起。主要和热带利什曼原虫分别导致动物源性和人源性 CL。巨噬细胞是利什曼原虫寄生虫感染的主要细胞，其产生一氧化氮（NO）的能力对寄生虫的消除至关重要。据我们所知，NO 对本地感染的作用以前从未被研究过。在这项研究中，我们评估了利什曼原虫主要和热带本地原代皮肤亲株对 J774 巨噬细胞产生 NO 的能力的体外调节作用，并确定了两种物种对外源性 NO 的敏感性。 方法：通过光学显微镜分析 J774 细胞系的感染性。通过 Griess 法测量巨噬细胞产生的 NO。使用 NO 供体通过 MTT 测定法评估两种菌株对 NO 的敏感性。 结果：我们的结果表明，主要和热带利什曼原虫菌株对巨噬细胞的感染率和每个巨噬细胞的寄生虫平均数量相似。虽然热带利什曼原虫显著抑制 LPS 和 IFN-γ刺激的 J774 巨噬细胞产生的 NO，但主要利什曼原虫没有影响。然而，两种本地亲代株的可溶性利什曼抗原（SLAs）均以剂量依赖性方式显著抑制 J774-巨噬细胞产生的 NO。最后，我们的结果表明，两种株的前鞭毛体和无鞭毛体均以剂量依赖性方式对 SNAP NO 供体敏感，尽管热带利什曼原虫表现出更高的敏感性。 结论：我们的结果表明，主要和热带利什曼原虫菌株调节巨噬细胞产生 NO 的能力存在差异。热带利什曼原虫抑制巨噬细胞产生 NO 的能力增加可能是由于其对 NO 供体的敏感性增加所致。我们的结果为热带利什曼原虫抑制巨噬细胞产生 NO 的能力增加提供了一种解释，这可能是由于摩洛哥 CL 的不同病理生理学所致。
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