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Peripheral Blood Mononuclear Cell Activation Induced by Leptospira interrogans Glycolipoprotein

机译：钩端螺旋体糖脂蛋白诱导的外周血单核细胞活化

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Leptospira interrogans glycolipoprotein (GLP) has been implicated in pathological and functional derangement seen in leptospirosis. The goal of this study was to evaluate GLP's ability to induce cellular activation, as assessed by cytokine production and expression of surface activation markers. GLP extracted from either pathogenic L. interrogans serovar Copenhageni or nonpathogenic Leptospira biflexa serovar Patoc (GLPp) was used to stimulate peripheral blood mononuclear cell cultures from healthy donors. Supernatant cytokine levels were measured by enzyme-linked immunosorbent assay. Expression of CD69 and HLA-DR on lymphocytes and monocytes, as well as lipopolysaccharide (LPS) binding, were measured by flow cytometry. At 6 h of incubation, GLP induced a significant rise in tumor necrosis factor alpha levels, which dropped progressively until 72 h of incubation. Interleukin-10 peak levels were obtained at between 24 and 48 h, with sustained levels until 72 h of incubation. The response magnitude was proportional to the GLP dose. CD69 expression on T lymphocytes and monocytes increased significantly, as did HLA-DR expression on monocytes. GLPp induced no CD69 or HLA-DR expression. GLP did not block biotinylated LPS binding to monocytes, suggesting that different pathways are used to induce cell activation. In conclusion, GLP induces cellular activation and may play a major role in the pathogenesis of leptospirosis.
机译：问号细螺旋体糖脂蛋白（GLP）与钩端螺旋体病的病理和功能紊乱有关。这项研究的目的是评估GLP诱导细胞活化的能力，通过细胞因子的产生和表面活化标记的表达来评估。从任何一种致病性 L提取的GLP。使用interrogans 血清型Copenhageni或非致病性 Leptospira biflexa 血清型Patoc（GLPp）刺激健康捐献者的外周血单核细胞培养。通过酶联免疫吸附测定法测量上清细胞因子水平。通过流式细胞术测量CD69和HLA-DR在淋巴细胞和单核细胞上的表达以及脂多糖（LPS）的结合。孵育6小时后，GLP诱导肿瘤坏死因子α水平显着上升，并逐渐下降直至孵育72 h。在24至48小时之间获得白介素10峰值水平，持续水平直至孵育72小时。反应幅度与GLP剂量成正比。 T淋巴细胞和单核细胞上的CD69表达显着增加，单核细胞上的HLA-DR表达也显着增加。 GLPp不诱导CD69或HLA-DR表达。 GLP并未阻止生物素化的LPS与单核细胞的结合，表明使用了不同的途径来诱导细胞活化。总之，GLP诱导细胞活化，并可能在钩端螺旋体病的发病机理中起主要作用。

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