体外培养的猪带绦虫囊尾蚴的排泄/分泌抗原(ES),以及抗ES单克隆抗体在神经囊虫病患者脑脊液样本中的抗原检测

N. M. Espíndola, A. Vaz, A. Pardini, I. Fernandes
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引用次数: 35

摘要

摘要采用无蛋白杂杂瘤培养基(PFHM)培养的方法,从带绦虫ORF株囊尾蚴中获得抗原。24-48h后观察新泡的萌发。排泄/分泌(ES)抗原(<20kDa的肽)培养48h后在培养基中恢复。囊液(VF)抗原与ES抗原的SDS-PAGE分析显示两者具有部分同源性。18- kda和14-kDa的ES肽被免疫VF抗原或全抗原制备的囊尾蚴绦虫制备的多克隆抗体所识别。神经囊虫患者血清或脑脊液(CSF)样本中存在的抗体也与ES肽反应。抗ES单克隆抗体检测了10例神经囊虫病患者脑脊液中的抗原,显示ES抗原与人感染猪囊尾蚴的抗原同源性。
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Excretory/secretory antigens (ES) from in-vitro cultures of Taenia crassiceps cysticerci, and use of an anti-ES monoclonal antibody for antigen detection in samples of cerebrospinal fluid from patients with neurocysticercosis
Abstract Antigens were obtained from cysticerci of the ORF strain of Taenia crassiceps, by culture of cysts in protein-free hybridoma medium (PFHM). Budding of new vesicles was observed after 24-48h. Excretory/secretory (ES) antigens (peptides of <20kDa) were recovered in the medium after culture for 48h. SDS-PAGE analysis of vesicular-fluid (VF) antigens (obtained by rupturing T. crassiceps cysticerci in PFHM) and the ES antigens indicated partial homology between the two preparations. ES peptides of 18- and 14-kDa were recognized by polyclonal antibodies produced in rabbits immunized either with the VF antigens or with a total-antigen preparation of T. solium cysticerci. Antibodies present in samples of serum or cerebrospinal fluid (CSF) from patients with neurocysticercosis also reacted with ES peptides. An anti-ES monoclonal antibody detected antigens in the CSF from 10 patients with neurocysticercosis, showing the antigenic homology of the ES antigens with those of T. solium cysticerci in human infections.
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