Instituto René Rachou, Fundação Oswaldo Cruz - FIOCRUZ, Belo Horizonte, Brazil.
Universidade Federal de São Paulo, UNIFESP, Diadema, Brazil.
Front Cell Infect Microbiol. 2019 Jul 10;9:240. doi: 10.3389/fcimb.2019.00240. eCollection 2019.
Lipophosphoglycan (LPG) is the major surface glycoconjugate having importance during the host-parasite interface. () displays a spectrum of clinical forms including: typical cutaneous leishmaniasis (TL), mucocutaneous (ML), and atypical lesions (AL). Those variations in the immunopathology may be a result of intraspecies polymorphisms in the parasite's virulence factors. In this context, we evaluated the role of LPG of strains originated from patients with different clinical manifestations and the sandfly vector. Six isolates of were used: M2903, RR051 and RR418 (TL), RR410 (AL), M15991 (ML), and M8401 (vector). LPGs were extracted and purified by hydrophobic interaction. Peritoneal macrophages from C57BL/6 and respective knock-outs (TLR2 and TLR-4) were primed with IFN-γ and exposed to different LPGs for nitric oxide (NO) and cytokine production (IL-1β, IL-6, IL-12, and TNF-α). LPGs differentially activated the production of NO and cytokines via TLR4. In order to ascertain if such functional variations were related to intraspecies polymorphisms in the LPG, the purified glycoconjugates were subjected to western blot with specific LPG antibodies (CA7AE and LT22). Based on antibody reactivity preliminary variations in the repeat units were detected. To confirm these findings, LPGs were depolymerized for purification of repeat units. After thin layer chromatography, intraspecies polymorphisms were confirmed especially in the type and/size of sugars branching-off the repeat units motif. In conclusion, different isolates of from different clinical forms and hosts possess polymorphisms in their LPGs that functionally affected macrophage responses.
脂磷壁酸 (LPG) 是主要的表面糖缀合物，在宿主-寄生虫界面具有重要作用。() 表现出一系列临床形式，包括：典型皮肤利什曼病 (TL)、黏膜皮肤利什曼病 (ML) 和非典型病变 (AL)。免疫病理学的这些变化可能是寄生虫毒力因素种内多态性的结果。在这种情况下，我们评估了来自不同临床表现和沙蝇媒介的患者的 LPG 菌株的作用。使用了 的六种分离株：M2903、RR051 和 RR418 (TL)、RR410 (AL)、M15991 (ML) 和 M8401 (媒介)。LPG 通过疏水相互作用提取和纯化。用 IFN-γ 对 C57BL/6 和各自的敲除体 (TLR2 和 TLR-4) 的腹腔巨噬细胞进行了初始处理，并将其暴露于不同的 LPG 中以产生一氧化氮 (NO) 和细胞因子 (IL-1β、IL-6、IL-12 和 TNF-α)。LPG 通过 TLR4 差异激活 NO 和细胞因子的产生。为了确定这种功能变化是否与 LPG 中的种内多态性有关，纯化的糖缀合物用特异性 LPG 抗体 (CA7AE 和 LT22) 进行了 Western blot。根据抗体反应性，初步检测到重复单元的差异。为了证实这些发现，将 LPG 进行解聚以纯化重复单元。在薄层层析后，特别是在重复单元的分支糖的类型和/或大小上，确认了种内多态性。总之，来自不同临床形式和宿主的 的不同分离株在其 LPG 中具有多态性，这些多态性在功能上影响了巨噬细胞的反应。
