Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, , P.O. Box 14155-6453, Iran.
Parasitol Res. 2014 Jan;113(1):229-37. doi: 10.1007/s00436-013-3648-0. Epub 2013 Oct 24.
Cryptosporidium parvum is a coccidian protozoan that causes diarrhea in immunocompromised humans and newborn animals. Billions of oocysts of C. parvum can be released from the infected calves and can contaminate the environment. The severity of the disease depends on the immunological status of the individual. Oocysts of Cryptosporidium are extremely resistant to many environmental stresses, and no effective disinfectant and curative agent against this organism is available. In our study, recombinant C. parvum P23 was prepared for application in the isolation and prevention of cryptosporidiosis. P23 is a glycoprotein that belongs to a family of protein of 23-27 kDa and is believed to be expressed in the different life stages of the parasite. Immunostaining analysis using the indirect fluorescent antibody test showed that P23 could be recognized on the surface of the oocysts. The antibody prepared in rabbit against P23 was bound to Sepharose 4B and used for the isolation of oocysts. The results showed that the prepared column was able to bind specifically only the oocysts. The effect of specific recombinant C. parvum IgY antibody against infection with C. parvum was examined in a mouse model. For this aim, purified egg yolk antibody prepared from immunized hens was used to analyze the protective effect of recombinant P23 specific antibody in immunosuppressed adult mice. The results showed more than 70% reduction in oocyst shedding after challenge with 1 × 10(4) oocysts. These results support previous studies of other investigators regarding the protective effect of P23 as an antigen against C. parvum infection and showed that it could be possible to design a passive immunization strategy against C. parvum based on the anti-P23 yolk antibody in animals and immunosuppressed humans.
微小隐孢子虫是一种引起免疫功能低下的人和新生动物腹泻的球虫原生动物。数以十亿计的微小隐孢子虫卵囊可从受感染的小牛中释放出来，并污染环境。疾病的严重程度取决于个体的免疫状态。微小隐孢子虫卵囊对许多环境压力具有极强的抵抗力，目前尚无针对该生物的有效消毒剂和治疗剂。在我们的研究中，制备了重组微小隐孢子虫 P23 以用于分离和预防隐孢子虫病。P23 是一种糖蛋白，属于 23-27 kDa 大小的蛋白家族，据信在寄生虫的不同生活阶段表达。使用间接荧光抗体试验的免疫染色分析表明，P23 可在卵囊表面被识别。用 P23 制备的兔抗血清与 Sepharose 4B 结合，用于分离卵囊。结果表明，制备的柱仅能特异性结合卵囊。在小鼠模型中检查了针对感染微小隐孢子虫的特异性重组微小隐孢子虫 IgY 抗体的效果。为此，用免疫母鸡制备的纯化卵黄抗体分析了重组 P23 特异性抗体在免疫抑制成年小鼠中的保护作用。结果显示，用 1×10(4)个卵囊攻毒后，卵囊脱落减少了 70%以上。这些结果支持了其他研究人员关于 P23 作为针对微小隐孢子虫感染的抗原的保护作用的先前研究，并表明可以根据动物和免疫抑制人类中的抗 P23 卵黄抗体设计针对微小隐孢子虫的被动免疫策略。
