Hornok S, Széll Z, Nieuwenhuijs J, Nieuwland M G, Cornelissen A W, Varga I
Department of Parasitology and Zoology, University of Veterinary Science, Budapest, Hungary.
Parasitol Res. 1999 Jan;85(1):71-7. doi: 10.1007/s004360050509.
The immunogenic properties of water-soluble and detergent-extracted components of Cryptosporidium baileyi oocysts were studied. Oocyst cytosol antigen (OCA) containing hydrophilic proteins was obtained by freeze-thaw cycles in liquid nitrogen. This was followed by Triton X-114 extraction of remaining oocyst fragments to dissolve membrane-bound proteins (TRE). The remainder of the pellet was solubilized with sodium dodecyl sulfate and treated with 2-mercaptoethanol to reduce disulfide-linked oocyst wall proteins (BME). The immune recognition of these three extracts was evaluated during the course of experimental cryptosporidiosis in chickens using ELISA, immunoblotting, and the lymphocyte stimulation test (LST). Four groups of chickens were infected at various times with different doses of C. baileyi and one group with the mammalian parasite C. parvum. Analysis of the data revealed that OCA proteins are well recognized by serum antibodies during the infection and to a limited extent by sera from chickens infected with C. parvum. Humoral responses of chicken groups to this antigen did not correlate well with the length of patency in contrast with its cellular recognition in LST. TRE gave lower values than OCA in both ELISA and LST, though it was still specifically recognized by samples from C. baileyi-infected chickens. Antibodies reacted aspecifically with BME, since only samples of birds which were immunocompetent at the time of their infection were able to recognize this extract as antigen. Immunoblotting revealed more specific components in OCA than in TRE or BME.
对贝利隐孢子虫卵囊的水溶性成分和经去污剂提取的成分的免疫原性进行了研究。通过在液氮中反复冻融获得了含有亲水性蛋白质的卵囊胞质溶胶抗原（OCA）。随后用 Triton X - 114 提取剩余的卵囊片段以溶解膜结合蛋白（TRE）。沉淀的其余部分用十二烷基硫酸钠溶解，并用 2 - 巯基乙醇处理以还原二硫键连接的卵囊壁蛋白（BME）。在鸡实验性隐孢子虫病过程中，使用酶联免疫吸附测定（ELISA）、免疫印迹和淋巴细胞刺激试验（LST）评估了这三种提取物的免疫识别情况。四组鸡在不同时间用不同剂量的贝利隐孢子虫感染，一组用哺乳动物寄生虫微小隐孢子虫感染。数据分析表明，在感染期间，OCA 蛋白能被血清抗体很好地识别，而感染微小隐孢子虫的鸡血清对其识别程度有限。与 LST 中的细胞识别情况相反，鸡群对该抗原的体液反应与排虫期长度的相关性不佳。在 ELISA 和 LST 中，TRE 的值均低于 OCA，不过感染贝利隐孢子虫的鸡的样本仍能特异性识别它。抗体与 BME 发生非特异性反应，因为只有在感染时具有免疫活性的鸡的样本才能将该提取物识别为抗原。免疫印迹显示，OCA 中比 TRE 或 BME 有更多的特异性成分。
