Asgari Qasem, Keshavarz Hossein, Shojaee Saeedeh, Motazedian Mohammad Hossein, Mohebali Mehdi, Miri Ramin, Mehrabani Davood, Rezaeian Mostafa
Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran ; Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Parasitol. 2013 Jul;8(3):367-75.
Based on recent studies, there are controversial reports on the capacity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capacity was evaluated by in vivo and in vitro experiments.
RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biological aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice.
All rats showed antibodies to Toxoplasma at titers ≥1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats' brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible bradyzoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice.
This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resistance to acidic condition, so this strain can be a candidate for future vaccine researches.
基于最近的研究,关于刚地弓形虫RH株形成组织包囊的能力存在有争议的报道。在本研究中,通过体内和体外实验对该能力进行了评估。
将RH株皮下接种到10只Wistar大鼠体内。1个月后,采集它们的血液、大脑、舌头和膈肌,并用MAT、PCR、病理学和生物测定方法进行评估。将寄生虫在细胞单层中培养。为了转变为缓殖子,将培养基的pH值改变为6.8。通过在酸性胃蛋白酶中孵育并接种到10只BALB/c小鼠体内来评估缓殖子的生物学特性。
所有大鼠的弓形虫抗体滴度均≥1:320,但在组织中未检测到DNA和组织包囊。将大鼠脑匀浆腹腔接种到BALB/c小鼠后,所有动物均未建立感染。在细胞培养期间,在酸性培养基中培养3至5天时,用PAS染色时可观察到囊样结构。在酸性胃蛋白酶培养基中孵育的包囊内可见的缓殖子不能杀死任何小鼠。
本研究证实伊朗的RH株已失去在大鼠体内形成组织包囊的潜力,并且在细胞培养中培养的缓殖子失去了对酸性条件的抗性,因此该菌株可作为未来疫苗研究的候选菌株。