Samuelson J C, Caulfield J P
J Cell Biol. 1985 May;100(5):1423-34. doi: 10.1083/jcb.100.5.1423.
Cercariae, the freshwater stage of Schistosoma mansoni infectious to man, are covered by a single unit membrane and an immunogenic glycocalyx. When cercariae penetrate the host skin, they transform to schistosomula by shedding tails, secreting mucous and enzymes, and forming microvilli over their surface. Here the loss of the glycocalyx from cercariae transforming in vitro was studied morphologically and biochemically. By scanning electron microscopy, the glycocalyx was a dense mesh composed of 15-30 nm fibrils that obscured spines on the cercarial surface. The glycocalyx was absent on organisms fixed without osmium and was partially lost when parasites aggregated in their own secretions before fixation. By transmission electron microscopy, a 1-2 microns thick mesh of 8-15-nm fibrils was seen on parasites incubated with anti-schistosomal antibodies or fixed in aldehydes containing tannic acid or ruthenium red. Cercariae transformed to schistosomula when tails were removed mechanically and parasites were incubated in saline. Within 5 min of transformation, organisms synchronously formed microvilli which elongated to 3-5 microns by 20 min and then were shed. However, considerable fibrillar material remained adherent to the double unit membrane surface of schistosomula. For biochemical labeling, parasites were treated with eserine sulfate, which blocked cercarial swimming, secretion, infectivity, and transformation to schistosomula. Material labeled by periodate oxidation and NaB3H4 was on the surface as shown by autoradiography and had an apparent molecular weight of greater than 10(6) by chromatography. Periodate-NaB3H4 glycocalyx had an isoelectric point of 5.0 +/- 0.4 and was precipitable with anti-schistosomal antibodies. More than 60% of the radiolabeled glycocalyx was released into the medium by transforming parasites in 3 h and was recovered as high molecular weight material. Parasites labeled with periodate and fluorescein-thiosemicarbazide and then transformed had a corona of fluorescence containing microvilli, much of which was shed onto the slide. Material on cercariae labeled by lodogen-catalyzed iodination was also of high molecular weight and was antigenic. In conclusion, the cercarial glycocalyx appears to be composed of acidic high molecular weight fibrils which are antigenic and incompletely cleared during transformation.
曼氏血吸虫的尾蚴是对人类具有感染性的淡水阶段,其表面覆盖着一层单位膜和一层免疫原性糖萼。当尾蚴穿透宿主皮肤时,它们通过甩掉尾巴、分泌黏液和酶以及在其表面形成微绒毛而转变为童虫。在此,我们从形态学和生物化学角度研究了体外转变过程中尾蚴糖萼的丢失情况。通过扫描电子显微镜观察,糖萼是由15 - 30纳米的纤维组成的致密网状结构,它掩盖了尾蚴表面的棘。未经锇固定的生物体上没有糖萼,而在固定前寄生虫聚集在自身分泌物中时,糖萼会部分丢失。通过透射电子显微镜观察,在用抗血吸虫抗体孵育或固定于含鞣酸或钌红的醛类中的寄生虫上,可见一层由8 - 15纳米纤维组成的1 - 2微米厚的网状结构。当机械去除尾巴并将寄生虫在盐水中孵育时,尾蚴转变为童虫。在转变的5分钟内,生物体同步形成微绒毛,到20分钟时微绒毛伸长至3 - 5微米,然后脱落。然而,相当多的纤维状物质仍附着在童虫的双单位膜表面。为进行生化标记,用硫酸毒扁豆碱处理寄生虫,该物质可阻止尾蚴游动、分泌、感染性以及向童虫的转变。经高碘酸盐氧化和NaB3H4标记的物质通过放射自显影显示在表面,通过色谱法测定其表观分子量大于10(6)。高碘酸盐 - NaB3H4糖萼的等电点为5.0±0.4,可被抗血吸虫抗体沉淀。超过60%的放射性标记糖萼在3小时内被转变的寄生虫释放到培养基中,并以高分子量物质形式回收。用高碘酸盐和荧光素 - 硫代半卡巴腙标记然后转变的寄生虫有一个含微绒毛的荧光晕,其中大部分脱落到载玻片上。用碘酰催化碘化标记的尾蚴上的物质也是高分子量且具有抗原性。总之,尾蚴糖萼似乎由酸性高分子量纤维组成,这些纤维具有抗原性且在转变过程中未被完全清除。