Wyatt C R, Perryman L E
Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164, USA.
Ann N Y Acad Sci. 2000;916:378-87. doi: 10.1111/j.1749-6632.2000.tb05316.x.
We have developed an assay to detect mucosally delivered antibody to Cryptosporidium parvum sporozoite antigens. We absorbed a recombinant 23-kD sporozoite protein to polystyrene microspheres, and used flow cytometry to detect, titer, and determine the isotype of antibody to p23 that was shed in the feces of experimentally infected calves. Noninoculated calves have low levels of mucosal antibody to p23, with IgG1 as the predominant isotype. Antibody titers rise in inoculated calves as the animals recover from cryptosporidiosis. A calf that was naturally protected from cryptosporidiosis had mucosal IgM and IgG1 isotype anti-p23 antibodies prior to challenge with C. parvum oocysts. Ten days after challenge, the calf had high titers of IgM, IgA, IgG1, and IgG2 anti-p23 antibodies. Together, the data show that this method can be used to assess mucosally delivered antibody to C. parvum.
我们开发了一种检测针对微小隐孢子虫子孢子抗原的黏膜递送抗体的检测方法。我们将一种重组23-kD子孢子蛋白吸附到聚苯乙烯微球上,并使用流式细胞术来检测、滴定以及确定在实验感染小牛粪便中排出的针对p23的抗体的亚型。未接种的小牛对p23的黏膜抗体水平较低,以IgG1作为主要亚型。随着动物从隐孢子虫病中恢复,接种小牛的抗体滴度会升高。一头天然对隐孢子虫病有抵抗力的小牛在受到微小隐孢子虫卵囊攻击之前就有黏膜IgM和IgG1亚型的抗p23抗体。攻击后十天,这头小牛有高滴度的IgM、IgA、IgG1和IgG2抗p23抗体。总之,数据表明该方法可用于评估针对微小隐孢子虫的黏膜递送抗体。