Shin Eun-Hee, Kim Sung-Bum, Nam Ho-Woo, Han Eun-Taek, Park Jae-Hwan, Ahn Hye-Jin, Chai Jong-Yil
Department of Parasitology, Seoul National University College of Medicine, and Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul 110-799, South Korea.
J Parasitol. 2004 Feb;90(1):161-6. doi: 10.1645/GE-3183.
Various monoclonal antibodies (mAbs) against Toxoplasma gondii RH tachyzoites were used for flow cytometric detection of intracellular parasites in murine splenic lymphocytes. Tg110 and Tg563 (reacting with the major surface protein SAG1), Tg505 (with another surface protein SAG2), Tg695 and Tg786 (with rhoptry proteins), Tg507, Tg621, and Tg317 (with dense granule proteins), Tg536 (with a microneme protein), and Tg685 (with a cytosol antigen) were the mAbs used. After an in vitro infection of lymphocytes with tachyzoites and reactions with the different mAbs, flow cytometry was performed using an indirect immunofluorescent technique. The proportions of whole infected lymphocytes and of each infected lymphocyte phenotype, CD4+ T cells, CD8+ T cells, and B cells, were determined, and their fluorescent intensities were quantified. The best reaction was seen when Tg110 or Tg695 was used as the mAbs. The results suggest that mAbs against surface or rhoptry proteins are highly useful for the flow cytometric detection of intracellular T. gondii in host cells.
使用多种抗刚地弓形虫RH速殖子的单克隆抗体（mAb），通过流式细胞术检测小鼠脾淋巴细胞内的寄生虫。所使用的单克隆抗体包括：Tg110和Tg563（与主要表面蛋白SAG1反应）、Tg505（与另一种表面蛋白SAG2反应）、Tg695和Tg786（与棒状体蛋白反应）、Tg507、Tg621和Tg317（与致密颗粒蛋白反应）、Tg536（与微线体蛋白反应）以及Tg685（与胞质溶胶抗原反应）。在用速殖子对淋巴细胞进行体外感染并与不同单克隆抗体反应后，采用间接免疫荧光技术进行流式细胞术检测。确定了整个感染淋巴细胞以及每种感染淋巴细胞表型（CD4 + T细胞、CD8 + T细胞和B细胞）的比例，并对其荧光强度进行了定量。当使用Tg110或Tg695作为单克隆抗体时，反应最佳。结果表明，针对表面或棒状体蛋白的单克隆抗体对于宿主细胞内刚地弓形虫的流式细胞术检测非常有用。
