Wiest P M, Kossmann R J, Tartakoff A M
Case Western Reserve University, Cleveland, Ohio.
Am J Trop Med Hyg. 1989 Jul;41(1):70-7.
The transformation of free-living cercariae to parasitic schistosomula of Schistosoma mansoni is accompanied by the formation of a multilaminate surface membrane which binds concanavalin A (Con A). This transition normally occurs with penetration of intact skin of the mammalian host, but can be accomplished by vortexing and incubating organisms at 37 degrees C in complex tissue culture medium (e.g., Earle's lactalbumin). We determined the minimal conditions that initiate surface maturation by monitoring surface changes by transmission electron microscopy and surface fluorescein-conjugated Con A labeling. Cercariae incubated in water for 2 hr at 37 degrees C retained their trilaminate surface and were non-fluorescent. After incubation in 116 mM NaCl for 3 hr at 23 degrees C, no organisms were covered by intense surface fluorescence; increasing the temperature to 37 degrees C resulted in 50% of the parasites becoming fluorescent. The addition of 26 mM NaHCO3 (pH 7.4) to parasites incubated in NaCl at 37 degrees C resulted in 98% of the organisms binding Con A. No significant difference in surface Con A binding was found between organisms incubated with the chelators EGTA or EDTA and controls. Mechanical stimulus by itself was not effective in stimulating surface maturation. The conditions that initiated surface maturation were elevation of temperature to 37 degrees C and NaHCO3-containing medium.
曼氏血吸虫自由生活的尾蚴转变为寄生性童虫的过程伴随着多层表面膜的形成,该表面膜能结合伴刀豆球蛋白A(Con A)。这种转变通常发生在哺乳动物宿主完整皮肤被穿透时,但也可通过在37℃下于复合组织培养基(如Earle氏乳白蛋白培养基)中涡旋和孵育虫体来实现。我们通过透射电子显微镜监测表面变化以及表面荧光素偶联的Con A标记来确定引发表面成熟的最低条件。尾蚴在37℃水中孵育2小时后仍保留其三层层状表面且无荧光。在23℃下于116 mM NaCl中孵育3小时后,没有虫体被强烈的表面荧光覆盖;将温度升至37℃导致50%的寄生虫产生荧光。向在37℃的NaCl中孵育的寄生虫中添加26 mM NaHCO₃(pH 7.4),结果98%的虫体结合Con A。在用螯合剂乙二醇双乙胺醚(EGTA)或乙二胺四乙酸(EDTA)孵育的虫体与对照之间,未发现表面Con A结合有显著差异。机械刺激本身对刺激表面成熟无效。引发表面成熟的条件是温度升至37℃和含NaHCO₃的培养基。