Lodes M J, Yoshino T P
Department of Zoology, University of Oklahoma, Norman 73019.
J Invertebr Pathol. 1990 Jul;56(1):75-85. doi: 10.1016/0022-2011(90)90147-x.
Excretory-secretory (E-S) products contained in supernatants from in vitro cultured Schistosoma mansoni primary sporocysts were assayed for their effects on the in vitro motility of Biomphalaria glabrata hemocytes. Both whole (unfractionated) and fractionated E-S products were tested in modified Boyden chemotaxis chambers. E-S product fractionation was accomplished using both membrane ultrafiltration (MF) and high-pressure liquid chromatography (HPLC). Transformation (Tr) products, but not those products released by 8-day sporocysts, significantly inhibited the random motility of hemocytes from an S. mansoni susceptible strain (M-Line) of B. glabrata. This activity was found in both high and low MF fractions of Tr but not in an intermediate MF fraction. In an effort to isolate the active component(s) of the high MF fraction, HPLC was used to separate components based on size exclusion. Although each of four HPLC fractions displayed some inhibitory activity, the greatest consistent activity was found in fraction 3, which was composed, predominantly, of a 108-kDa protein. In contrast to the response of M-Line cells to Tr E-S products, the motility of hemocytes from an S. mansoni-resistant strain (10-R2-OK) of B. glabrata was not significantly reduced from controls. The high MF fraction, however, elicited a slight positive chemokinetic response, while the low MF fraction reduced 10-R2-OK hemocyte motility slightly but not significantly. While three HPLC fractions significantly reduced 10-R2-OK hemocyte motility, this effect was significantly less than that produced by the same HPLC fractions on M-Line hemocyte motility. These data suggest that S. mansoni sporocyst Tr E-S products differentially affect the random motility of M-Line and 10-R2-OK snail hemocytes. Although the significance of this differential effect on the in vivo defenses of B. glabrata is not known, it could be important in the host-parasite interaction which leads to either resistance or susceptibility.
检测体外培养的曼氏血吸虫初代子孢子上清液中所含的排泄分泌（E-S）产物对光滑双脐螺血细胞体外运动能力的影响。完整（未分级）和分级的E-S产物均在改良的博伊登趋化性小室中进行测试。E-S产物分级采用膜超滤（MF）和高压液相色谱（HPLC）两种方法。转化（Tr）产物，而非8日龄子孢子释放的产物，能显著抑制光滑双脐螺曼氏血吸虫易感品系（M系）血细胞的随机运动能力。在Tr的高、低MF级分中均发现了这种活性，而在中间MF级分中未发现。为了分离高MF级分中的活性成分，采用HPLC根据尺寸排阻分离成分。尽管四个HPLC级分均表现出一定的抑制活性，但最大且一致的活性出现在级分3中，该级分主要由一种108 kDa的蛋白质组成。与M系细胞对Tr E-S产物的反应不同，光滑双脐螺曼氏血吸虫抗性品系（10-R2-OK）血细胞的运动能力与对照组相比没有显著降低。然而，高MF级分引起了轻微的正向化学动力学反应，而低MF级分则略微降低了10-R2-OK血细胞运动能力，但不显著。虽然三个HPLC级分显著降低了10-R2-OK血细胞运动能力，但这种作用明显小于相同HPLC级分对M系血细胞运动能力的影响。这些数据表明，曼氏血吸虫子孢子Tr E-S产物对M系和10-R2-OK蜗牛血细胞的随机运动能力有不同影响。尽管这种差异效应在光滑双脐螺体内防御中的意义尚不清楚，但它在导致抗性或易感性的宿主-寄生虫相互作用中可能很重要。
