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在感染微小泰勒虫的牛淋巴细胞中寄生虫抗原的免疫组织化学检测

Immunohistochemical detection of parasite antigens in Theileria parva-infected bovine lymphocytes.

作者信息

Honda Y, Matsubara Y, Morzaria S, McKeever D

机构信息

International Livestock Research Institute (ILRI), Nairobi, Kenya.

出版信息

Vet Parasitol. 1998 Dec 31;80(2):137-47. doi: 10.1016/s0304-4017(98)00184-8.

Abstract

Theileria parva is the causal agent of East Coast fever (ECF), a fatal disease of cattle characterized by pyrexia, transient lymphadenopathy and panleukopenia. We have evaluated monoclonal antibodies (mAb) against three distinct antigens (p67, PIM and p32) of the parasite as immunohistological reagents for monitoring the kinetics of infection in cattle. Bovine lymphocytes were stained with the mAb at various intervals after infection in vitro and in vivo. The p67 sporozoite surface antigen was detected in only a small percentage of both, in vitro and in vivo infected cells. In contrast, expression of the polymorphic immunodominant molecule (PIM) of the parasite proved a useful indicator of infection and staining was correlated with the results of Giemsa analysis. PIM was detected from day 3 in in vitro-infected cells, but was not detected until day 5 in vivo after challenge with a 70% lethal dose of stabilized sporozoite. The p32 antigen was expressed only late in infection in vivo and its expression was associated with the development of merozoites. Less than 20% of in vitro-infected cells expressed p32. The immunohistochemical staining with anti-PIM mAb was found to be a useful tool for analysis of T. parva infection kinetics in cattle.

摘要

泰勒虫是东海岸热(ECF)的病原体,东海岸热是一种牛的致命疾病,其特征为发热、短暂性淋巴结病和全血细胞减少。我们评估了针对该寄生虫三种不同抗原(p67、PIM和p32)的单克隆抗体(mAb),作为监测牛感染动力学的免疫组织化学试剂。在体外和体内感染后的不同时间间隔,用该单克隆抗体对牛淋巴细胞进行染色。p67子孢子表面抗原在体外和体内感染的细胞中仅在一小部分细胞中被检测到。相比之下,该寄生虫多态性免疫显性分子(PIM)的表达被证明是感染的一个有用指标,且染色结果与吉姆萨分析结果相关。在体外感染的细胞中,从第3天开始检测到PIM,但在用70%致死剂量的稳定化子孢子攻击后,体内直到第5天才检测到PIM。p32抗原仅在体内感染后期表达,其表达与裂殖子的发育有关。体外感染的细胞中表达p32的细胞不到20%。发现用抗PIM单克隆抗体进行免疫组织化学染色是分析牛泰勒虫感染动力学的一种有用工具。

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