Ruppel A, McLaren D J
Exp Parasitol. 1986 Oct;62(2):223-36. doi: 10.1016/0014-4894(86)90027-5.
The human complement component C3b is known to bind in vitro to the surfaces of all developmental stages of schistosomes as a consequence of complement activation by the alternative pathway. C3b bound to Schistosoma mansoni parasites has now been used in combination with fluorescent labeled antibodies against C3b to label the surfaces of living schistosomes. Binding of complement components and labeled antibodies to adult schistosomes rendered their surface membrane homogeneously fluorescent. At the ultrastructural level, the label was seen as a dense deposit lying on the tegumental membrane. Surface damage was not observed in labeled adults by electron microscopy. Fluorescent schistosomes were cultured in vitro for periods of up to 2 weeks, during which time the parasites remained fully viable and their surface membrane was still fluorescent. The electron dense deposit persisted, and tegumental damage at the electron microscope level was minimal or absent. Consequently, adult schistosomes would seem able to survive in vitro in the absence of rapid and general turnover of their surface membrane. Loss of fluorescence was observed consistently only at the anterior end of the parasite, including the suckers, a finding which indicates that membrane turnover may occur at different rates on different parts of the body. Fluorescent 3-week-old juveniles and 6-day-old lung stage parasites were cultured under the same conditions with similar results: they remained viable and fluorescent for at least 2 weeks. Results with skin schistosomula were different in the sense that many worms died during culture, and those which survived lost large parts of their fluorescent surface. A few of the surviving and fluorescent schistosomula developed the elongate shape typical of lung stage parasites. Fluorescent viable skin schistosomula were injected intravenously into mice and subsequently recovered from the lungs after varying periods. Fluorescence was lost in a patchy way within a few minutes from some individuals and within several hours from most of the worms. These data permit the following conclusions: C3b is a suitable tracer for membrane renewal in all developmental stages of schistosomes. Very slow membrane renewal in vitro and very rapid renewal in vivo are both compatible with parasite survival.
已知人类补体成分C3b在体外可通过替代途径激活补体后与血吸虫成虫各个发育阶段的表面相结合。与曼氏血吸虫结合的C3b目前已与抗C3b荧光标记抗体结合使用,用于标记活血吸虫的表面。补体成分及标记抗体与成虫血吸虫的结合使它们的表面膜呈现均匀的荧光。在超微结构水平上,标记物表现为位于皮层膜上的致密沉积物。电子显微镜观察未发现标记成虫表面有损伤。荧光血吸虫在体外培养长达2周,在此期间,寄生虫仍完全存活,其表面膜仍有荧光。电子致密沉积物持续存在,电子显微镜水平下的皮层损伤最小或不存在。因此,成虫血吸虫似乎能够在体外存活,而无需其表面膜快速且普遍地更新。仅在寄生虫前端(包括吸盘)持续观察到荧光损失,这一发现表明膜更新在身体不同部位可能以不同速率发生。荧光标记的3周龄幼虫和6日龄肺部阶段寄生虫在相同条件下培养,结果相似:它们至少2周内仍存活且有荧光。皮肤期血吸虫的结果不同,许多虫体在培养过程中死亡,存活下来的虫体荧光表面大部分消失。少数存活且有荧光的皮肤期血吸虫发育出典型的肺部阶段寄生虫的细长形状。将荧光标记的活皮肤期血吸虫静脉注射到小鼠体内,随后在不同时间段后从肺部回收。几分钟内,一些个体的荧光呈斑片状消失,大多数虫体在数小时内荧光消失。这些数据可得出以下结论:C3b是血吸虫所有发育阶段膜更新的合适示踪剂。体外非常缓慢的膜更新和体内非常快速的膜更新均与寄生虫存活相容。