Skilton R A, Musoke A J, Wells C W, Yagi Y, Nene V, Spooner P R, Gachanja J, Osaso J, Bishop R P, Morzaria S P
International Livestock Research Institute (ILRI), Nairobi, Kenya.
Parasitology. 2000 Jun;120 ( Pt 6):553-64. doi: 10.1017/s0031182099005934.
Previous studies using monoclonal antibody (mAb) 4C9 specific for a 32 kDa antigen (p32) of Theileria parva demonstrated expression of the antigen on the surface of the sporozoite, making it a potential antigen for sporozoite neutralization. A full-length cDNA encoding the major merozoite/piroplasm surface antigen (mMPSA) of T. parva was cloned and expressed in bacteria. The expressed product reacted strongly with mAb 4C9, demonstrating identity between the p32 and mMPSA of T. parva. Using immunoblot analysis and immunoelectron microscopy with mAb 4C9 it was shown that the mMPSA is a major antigen of the merozoite and piroplasm at the cell surface, while lower levels of antigen are expressed in the sporozoite and schizont stages. Upregulation of the mMPSA occurs at merogony and can be induced by culturing schizont-infected lymphocytes at 42 degrees C. Recombinant mMPSA of T. parva induced high titres of specific antibodies in cattle but failed to confer protection against a T. parva sporozoite stabilate challenge. The pre-challenge sera also failed to neutralize infectivity of sporozoites in an in vitro assay. Possible reasons for the lack of parasite neutralization in vivo and in vitro are discussed.
先前使用针对微小泰勒虫32 kDa抗原(p32)的单克隆抗体(mAb)4C9进行的研究表明,该抗原在子孢子表面表达,使其成为子孢子中和的潜在抗原。克隆了编码微小泰勒虫主要裂殖子/梨形虫体表抗原(mMPSA)的全长cDNA,并在细菌中表达。表达产物与mAb 4C9强烈反应,证明微小泰勒虫的p32和mMPSA具有同一性。使用mAb 4C9进行免疫印迹分析和免疫电子显微镜检查表明,mMPSA是裂殖子和梨形虫在细胞表面的主要抗原,而在子孢子和裂殖体阶段表达的抗原水平较低。mMPSA在裂体增殖时上调,并且可以通过在42℃培养感染裂殖体的淋巴细胞来诱导。微小泰勒虫的重组mMPSA在牛中诱导了高滴度的特异性抗体,但未能对微小泰勒虫子孢子稳定株攻击提供保护。攻毒前血清在体外试验中也未能中和子孢子的感染性。讨论了体内和体外缺乏寄生虫中和作用的可能原因。