Cox G N, Shamansky L M, Boisvenue R J
Synergen, Inc., Boulder, CO 80301.
Mol Biochem Parasitol. 1989 Oct;36(3):233-41. doi: 10.1016/0166-6851(89)90171-0.
Cuticular surface antigens of the XL3 and L4 stages of Haemonchus contortus have been studied by surface labeling and immunological techniques. Live worms were labeled with 125I and extracted with sodium dodecyl sulfate (SDS) followed by SDS + 2-mercaptoethanol. The SDS-soluble surface proteins of XL3s and L4s were found to consist of relatively few major species. The pattern of labeled polypeptides was distinctive for each developmental stage. These proteins are refractory to digestion by bacterial collagenase. Several of the proteins are glycosylated. Further extraction of labeled worms with SDS + 2-mercaptoethanol solubilized additional labeled proteins that appeared to be primarily collagens. Rabbit antisera prepared against native XL3 and L4-cuticles reacted strongly with the surfaces of live worms in immunofluorescence assays. In contrast, antisera prepared against SDS-extracted cuticles reacted weakly or not at all with live worms in similar experiments. Rabbit antisera prepared against adult cuticles failed to react with live XL3s or L4s. These studies suggest that the major surface antigens of XL3s and L4s are solubilized by SDS and that there are different antigens present on the cuticular surfaces of XL3s, L4s and adults. Stage-specificity in cuticular surface proteins may contribute to the successful parasitic lifestyle of this nematode.
利用表面标记和免疫学技术对捻转血矛线虫XL3和L4阶段的角质层表面抗原进行了研究。将活虫用¹²⁵I标记,并用十二烷基硫酸钠(SDS)提取,随后用SDS + 2-巯基乙醇提取。发现XL3和L4的SDS可溶性表面蛋白由相对较少的主要种类组成。每个发育阶段标记多肽的模式都很独特。这些蛋白质对细菌胶原酶消化具有抗性。其中几种蛋白质是糖基化的。用SDS + 2-巯基乙醇对标记的虫体进行进一步提取,可溶解额外的标记蛋白,这些蛋白似乎主要是胶原蛋白。在免疫荧光试验中,针对天然XL3和L4角质层制备的兔抗血清与活虫表面强烈反应。相比之下,在类似实验中,针对SDS提取的角质层制备的抗血清与活虫反应较弱或根本不反应。针对成虫角质层制备的兔抗血清与活的XL3或L4不发生反应。这些研究表明,XL3和L4的主要表面抗原可被SDS溶解,并且XL3、L4和成虫的角质层表面存在不同的抗原。角质层表面蛋白的阶段特异性可能有助于这种线虫成功的寄生生活方式。