由杜氏利什曼原虫引起的人类皮肤利什曼病中的细胞因子和表型细胞特征。

BACKGROUND: The innate immune mediators are likely to influence the clinical phenotype of leishmaniasis by primary responses which limit or facilitate the spread of the parasite, as well as by modulating adaptive immunity. This study investigated the response of key innate immune cells in a focus which regularly reports localised cutaneous leishmaniasis (LCL) caused by Leishmania donovani, a species which typically causes visceral disease. METHODS: Peripheral blood mononuclear cell (PBMC) derived macrophages and dendritic cells from patients with LCL and healthy controls from endemic and non-endemic areas, were stimulated with soluble Leishmania antigen (SLA). Inflammatory mediators produced by macrophages (TNF-α/TGF-β/IL-10, ELISA; NO, Griess method) and dendritic cells (IL-12p70, IL-10, flowcytometry) and macrophage expression of surface markers of polarization, activation and maturation (flowcytometry) were determined at 24h, 48h and 72h and compared. Study was conducted prospectively from 2015-2019. RESULTS: Patient derived macrophages and dendritic cells produced higher levels of both pro and anti-inflammatory mediators compared to controls (p<0.05) with the best discrimination for active disease observed at 72h. Data demonstrated an early activation of macrophages and a subsequent pro-inflammatory bias, as indicated by temporal profiles of TNF-α/TGF-β and TNF-α/IL-10 ratios and higher proportions of classical (M1) macrophages. Higher TGF-β levels were observed in cells from patients with ulcerated or persistent lesions. Immune responses by cells derived from controls in endemic and non-endemic regions did not differ significantly from each other. CONCLUSIONS: The overall immunophenotypic profile suggests that LCL observed in the country is the result of a balancing immune response between pro-inflammatory and regulatory mediators. The mediators which showed distinct profiles in patients warrant further investigation as potential candidates for immunotherapeutic approaches. A comparison with visceral leishmaniasis caused by the same species, would provide further evidence on the differential role of these mediators in the resulting clinical phenotype.

背景：先天免疫介质可能通过限制或促进寄生虫传播的原发性反应，以及通过调节适应性免疫来影响利什曼病的临床表型。本研究调查了在一个经常报告由利什曼原虫引起的局限性皮肤利什曼病（LCL）的焦点中关键先天免疫细胞的反应，该物种通常导致内脏疾病。

方法：来自利什曼病患者和来自流行地区和非流行地区的健康对照的外周血单核细胞（PBMC）衍生的巨噬细胞和树突状细胞用可溶性利什曼抗原（SLA）刺激。巨噬细胞（TNF-α/TGF-β/IL-10，ELISA；NO，Griess 法）和树突状细胞（IL-12p70、IL-10、流式细胞术）产生的炎症介质以及巨噬细胞表面极化、激活和成熟的标记物的表达（流式细胞术）在 24 小时、48 小时和 72 小时进行测定并进行比较。研究于 2015-2019 年进行前瞻性。

结果：与对照相比，患者来源的巨噬细胞和树突状细胞产生了更高水平的促炎和抗炎介质（p<0.05），在 72 小时观察到对活动性疾病的最佳区分。数据表明巨噬细胞的早期激活和随后的促炎偏向，如 TNF-α/TGF-β 和 TNF-α/IL-10 比值的时间曲线和更高比例的经典（M1）巨噬细胞所示。在溃疡或持续病变患者的细胞中观察到较高的 TGF-β 水平。来自流行地区和非流行地区的对照细胞的免疫反应彼此之间没有显着差异。

结论：整体免疫表型谱表明，该国观察到的 LCL 是促炎和调节介质之间平衡免疫反应的结果。在患者中表现出不同特征的介质值得进一步研究，作为免疫治疗方法的潜在候选物。与由同一物种引起的内脏利什曼病进行比较，将为这些介质在产生的临床表型中的差异作用提供进一步证据。