Knight M, Simpson A J, Payares G, Chaudri M, Smithers S R
EMBO J. 1984 Jan;3(1):213-9. doi: 10.1002/j.1460-2075.1984.tb01786.x.
Messenger RNA has been extracted from all stages of the life cycle of the parasitic multicellular helminth Schistosoma mansoni. In vitro translation of these mRNA preparations in rabbit reticulocyte lysates yielded in each case a large number of polypeptides. Immunoprecipitation of translation products either by serum from immune mice or from human patients demonstrated that relatively few, approximately 10, polypeptides are recognised as antigens. Two of the in vitro synthesised antigens, of mol. wts. 22 000 and 14 000, were demonstrated to correspond to schistosomula surface antigens. The expression of these antigens may show stage specificity. Both are readily detected from adult and sporocyst translation products, neither from schistosomula and only the 22 000 antigen from miracidia. This is an unexpected finding since similar polypeptide antigens occur on the surface of schistosomula. These results indicate that not only are schistosomula surface antigens preformed at the preceding sporocyst stage, i.e., within the snail host, but they also remain invariant throughout the life cycle in the vertebrate host. Two other prominent schistosomula surface antigens of mol. wts. 38 000 and 32 000, were not recognised amongst cell-free translation products directed by RNA from any life cycle stage. The demonstration that at least two schistosomula surface antigens are detectable amongst adult mRNA cell-free translation products demonstrates the feasibility of identifying the genes encoding them in cDNA libraries from adult worm mRNA.
已从寄生性多细胞蠕虫曼氏血吸虫生命周期的各个阶段提取了信使核糖核酸(mRNA)。在兔网织红细胞裂解物中对这些mRNA制剂进行体外翻译,每种情况下都产生了大量多肽。用免疫小鼠血清或人类患者血清对翻译产物进行免疫沉淀表明,相对较少的大约10种多肽被识别为抗原。两种体外合成的抗原,分子量分别为22000和14000,被证明对应于童虫表面抗原。这些抗原的表达可能具有阶段特异性。在成虫和孢蚴的翻译产物中都能很容易地检测到这两种抗原,在童虫中都检测不到,而在毛蚴中只能检测到分子量为22000的抗原。这是一个意想不到的发现,因为类似的多肽抗原出现在童虫表面。这些结果表明,不仅童虫表面抗原在之前的孢蚴阶段即蜗牛宿主体内就已预先形成,而且它们在脊椎动物宿主的整个生命周期中都保持不变。另外两种突出的童虫表面抗原,分子量分别为38000和32000,在来自任何生命周期阶段的RNA指导的无细胞翻译产物中均未被识别。在成虫mRNA的无细胞翻译产物中能够检测到至少两种童虫表面抗原,这一结果证明了从成虫mRNA的cDNA文库中鉴定编码它们的基因的可行性。