Department of Pathobiological Sciences, University of Wisconsin, School of Veterinary Medicine, Madison, WI 53706, USA.
Exp Parasitol. 2013 Jan;133(1):28-36. doi: 10.1016/j.exppara.2012.10.002. Epub 2012 Oct 22.
Host lectin-like recognition molecules may play an important role in innate resistance in Biomphalaria glabrata snails to larval schistosome infection, thus implicating parasite-expressed glycans as putative ligands for these lectin receptors. While host lectins may utilize specific glycan structures for parasite recognition, it also has been hypothesized that the parasite may use this system to evade immune detection by mimicking naturally-expressed host glycans, resulting in reduced immunorecognition capacity. By employing immunocytochemical (ICC) and Western blot assays using schistosome glycan-specific monoclonal antibodies (mABs) we sought to identify specific glycan epitopes (glycotopes) shared in common between larval Schistosoma mansoni and B. glabrata hemocytes, the primary immune effector cells in snails. Results confirmed the presence of selected larval glycotopes on subpopulations of hemocytes by ICC and association with numerous hemocyte proteins by Western blot analyses, including a trimannosyl core N-glycan (TriMan), and two fucosylated lacdiNAc (LDN) variants, F-LDN and F-LDN-F. Snail strain differences were seen in the prevalence of constitutively expressed F-LDN on hemocytes, and in the patterns of protein immunoreactivity with these mABs. In contrast, there was little to no hemocyte reactivity with mABs for Lewis X (LeX), LDN, LDN-F or LDN-DF. When intact hemocytes were exposed to larval transformation products (LTPs), distinct cell subpopulations displayed weak (LeX, LDN-DF) to moderate (LDN, LDN-F) glycotope reactivity by ICC, including snail strain differences in the prevalence of LDN-reactive cellular subsets. Far-Western blot analyses of the hemocytes following exposure to larval transformation proteins (LTPs) also revealed multiple mAB-reactive hemocyte protein bands for LeX, LDN, LDN-F, and LDN-DF. These results demonstrate the existence of complex patterns of shared larval glycan constitutively expressed on hemocytes and their proteins, as well as the ability of hemocytes to acquire shared glycans by the selective binding of parasite-released LTP. Unraveling the functional significance of these naturally expressed and acquired shared glycans on specific hemocyte populations represents an important challenge for future investigations.
宿主凝集素样识别分子可能在豆螺对幼虫期血吸虫感染的先天抗性中发挥重要作用,因此寄生虫表达的糖可能是这些凝集素受体的假定配体。虽然宿主凝集素可能利用特定的糖结构来识别寄生虫,但也有人假设寄生虫可能利用这种系统通过模拟天然表达的宿主糖来逃避免疫检测,从而降低免疫识别能力。通过使用针对血吸虫糖的单克隆抗体 (mAB) 的免疫细胞化学 (ICC) 和 Western blot 分析,我们试图鉴定幼虫期曼氏血吸虫和豆螺血淋巴细胞之间共同存在的特定糖表位 (糖基),血淋巴细胞是螺类的主要免疫效应细胞。结果通过 ICC 证实了选择的幼虫糖基在血淋巴细胞亚群中的存在,并通过 Western blot 分析与许多血淋巴细胞蛋白相关联,包括三甘露糖核心 N-聚糖 (TriMan) 和两种岩藻糖基化 lacdiNAc (LDN) 变体,F-LDN 和 F-LDN-F。在血淋巴细胞中组成型表达的 F-LDN 的流行率以及与这些 mAB 的蛋白免疫反应模式方面,观察到螺种差异。相比之下,mAB 对 Lewis X (LeX)、LDN、LDN-F 或 LDN-DF 几乎没有或没有血淋巴细胞反应。当完整的血淋巴细胞暴露于幼虫转化产物 (LTP) 时,通过 ICC,不同的细胞亚群显示出较弱 (LeX、LDN-DF) 到中等 (LDN、LDN-F) 的糖基反应性,包括在 LDN 反应性细胞亚群的流行率方面存在螺种差异。幼虫转化蛋白 (LTP) 暴露后对血淋巴细胞进行 Far-Western blot 分析,也揭示了 LeX、LDN、LDN-F 和 LDN-DF 的多个 mAB 反应性血淋巴细胞蛋白带。这些结果表明,在血淋巴细胞及其蛋白上存在复杂的幼虫糖基组成型表达模式,以及血淋巴细胞通过选择性结合寄生虫释放的 LTP 获得共同糖基的能力。阐明这些在特定血淋巴细胞群体上自然表达和获得的共同糖的功能意义,是未来研究的重要挑战。