Marinkovic Emilija, Djokic Radmila, Lukic Ivana, Filipovic Ana, Inic-Kanada Aleksandra, Kosanovic Dejana, Gavrovic-Jankulovic Marija, Stojanovic Marijana
Department of Research and Development; Institute of Virology, Vaccines and Sera-TORLAK; Belgrade, Serbia.
OCUVAC-Center of Ocular Inflammation and Infection, Laura Bassi Centres of Expertise; Center for Pathophysiology, Infectiology and Immunology; Medical University of Vienna; Vienna, Austria.
PLoS One. 2017 Feb 24;12(2):e0172469. doi: 10.1371/journal.pone.0172469. eCollection 2017.
We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 μg to 10 μg to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNFα secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGFβ and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFα and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.
我们证明,一种重组香蕉凝集素(rBanLec),其结构特征和生理影响与天然对应物的报道高度相似,可结合小鼠腹膜巨噬细胞并特异性调节其功能特性。通过使用浓度范围为1μg至10μg的rBanLec刺激来自BALB/c和C57BL/6小鼠的驻留腹膜巨噬细胞(RMs)和巯基乙酸盐诱导的腹膜巨噬细胞(TGMs),我们发现rBanLec刺激的效果不仅取决于其浓度,还取决于巨噬细胞的功能状态及其遗传背景。rBanLec以正剂量依赖性方式促进来自BALB/c和C57BL/6小鼠的TGMs增殖,而其对RMs的促有丝分裂影响显著较低(BALB/c小鼠)或无法检测到(C57BL/6小鼠)。在所有腹膜巨噬细胞中,无论其类型和遗传背景如何,rBanLec均以正剂量依赖性方式增强IL-10的分泌。rBanLec对来自BALB/c和C57BL/6的RMs的刺激导致促炎表型呈正剂量依赖性促进(NO产生、IL-12和TNFα分泌增强,精氨酸酶活性降低)。在来自C57BL/6小鼠的TGMs中也观察到向促炎表型的正剂量依赖性偏向。然而,rBanLec刺激的增强促进来自BALB/c小鼠的TGMs向抗炎谱的偏向(NO产生和IL-12分泌减少,精氨酸酶活性以及TGFβ和IL-4分泌增强)。此外,我们确定rBanLec结合TLR2和CD14的寡糖结构,并且通过这些受体阻断信号传导会显著损害BALB/c巨噬细胞中TNFα和NO的产生。由于rBanLec刺激的结果取决于rBanLec浓度以及其靶细胞的功能特性及其遗传背景,因此需要进一步研究以探讨其在生理和特定病理条件下的作用。