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抗三磷酸核苷水解酶-II的单克隆抗体可减少刚地弓形虫的复制。

Monoclonal antibodies against nucleoside triphosphate hydrolase-II can reduce the replication of Toxoplasma gondii.

作者信息

Tan Feng, Hu Xin, Pan Chang-Wang, Ding Jie-Qiong, Chen Xiao-Guang

机构信息

Department of Parasitology, School of Public Health and Tropical Medicine, Southern Medical University, Guangdong, PR China.

出版信息

Parasitol Int. 2010 Jun;59(2):141-6. doi: 10.1016/j.parint.2009.12.007. Epub 2010 Jan 6.

Abstract

Nucleoside triphosphate hydrolase (NTPase) is an abundant protein secreted by the obligate protozoan parasite Toxoplasma gondii, which has a wide specificity toward NTP. In the present study, two monoclonal antibodies (mAbs, MNT1 and MNT2) against recombinant T. gondii NTPase-II (rTgNTPase-II) were developed. Western blot analysis displayed that these two mAbs can recognize specifically rTgNTPase-II as well as a 63kDa molecule in tachyzoites soluble antigens that corresponded to native NTPase-II. T. gondii tachyzoites pretreated with two mAbs were observed under Confocal Laser Microscope and a specific reaction was displayed on tachyzoites after indirect fluorescence antibody test (IFAT). When COS-7 cells were co-cultured with tachyzoites pretreated with two mAbs, the number of intracellular parasites per infected cell was significantly decreased compared with the control. Furthermore, incubation of T. gondii tachyzoites with two mAbs can inhibit NTPase activity in the presence of dithiothreitol, which hinted that the reduction of tachyzoite replication might be owing to the inhibition of NTPase-II by the mAbs. The passive immunization test indicated that the transferred mAbs can significantly prolong the survival time of challenge infected mice. Taken together, we concluded that the mAbs against NTPase-II can reduce the replication of T. gondii and have a crucial effect on the protection of host from T. gondii infection.

摘要

核苷三磷酸水解酶(NTPase)是专性原生动物寄生虫刚地弓形虫分泌的一种丰富蛋白质,它对NTP具有广泛的特异性。在本研究中,制备了两种针对重组刚地弓形虫NTPase-II(rTgNTPase-II)的单克隆抗体(mAb,MNT1和MNT2)。蛋白质免疫印迹分析显示,这两种单克隆抗体能够特异性识别rTgNTPase-II以及速殖子可溶性抗原中与天然NTPase-II相对应的一个63kDa分子。用这两种单克隆抗体预处理的刚地弓形虫速殖子在共聚焦激光显微镜下观察,间接荧光抗体试验(IFAT)后速殖子上呈现出特异性反应。当COS-7细胞与用这两种单克隆抗体预处理的速殖子共培养时,与对照相比,每个感染细胞内的寄生虫数量显著减少。此外,用这两种单克隆抗体孵育刚地弓形虫速殖子在二硫苏糖醇存在下可抑制NTPase活性,这表明速殖子复制的减少可能是由于单克隆抗体对NTPase-II的抑制作用。被动免疫试验表明,转移的单克隆抗体可显著延长攻击感染小鼠的存活时间。综上所述,我们得出结论,针对NTPase-II的单克隆抗体可减少刚地弓形虫的复制,并对保护宿主免受刚地弓形虫感染具有关键作用。

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