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使用实时定量聚合酶链反应(Real time QPCR)测定经排泄分泌抗原免疫后小鼠弓形虫病不同组织中的寄生虫负荷。

Determination of parasitic load in different tissues of murine toxoplasmosis after immunization by excretory-secretory antigens using Real time QPCR.

作者信息

Daryani Ahmad, Sharif Mehdi, Dadimoghaddam Yousef, Souteh Mohammad Bagher Hashemi, Ahmadpour Ehsan, Khalilian Alireza, Sarvi Shahabeddin, Farazmand Touraj, Kalani Hamed, Rasouli Mehdi

机构信息

Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran; Parasitology and Mycology Department, Sari Medical School, Mazandaran University of Medical Sciences, Sari, Iran.

Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran; Parasitology and Mycology Department, Sari Medical School, Mazandaran University of Medical Sciences, Sari, Iran.

出版信息

Exp Parasitol. 2014 Aug;143:55-9. doi: 10.1016/j.exppara.2014.05.008. Epub 2014 May 20.

Abstract

Excretory-secretory antigens (ESAs) of Toxoplasma gondii are one of the candidates for immunization against toxoplasmosis. For evaluation of immunization, we determined the kinetics of the distribution of Toxoplasma and parasite load in different tissues of mice immunized by ESAs. In this experimental study, 36 mice in case (n=18) and control (n=18) groups were immunized with ESAs and PBS, respectively. After 2weeks, mice were challenged intraperitoneally with Toxoplasma virulent RH strain. Blood and different tissues (brain, spleen, liver, heart, kidney, and muscle) were collected daily after challenge (1, 2, 3 and last day before death). Parasite load was calculated using Real time QPCR targeted at the B1 gene. ESAs as vaccine in different tissues showed various effects. However, infected mice which received the vaccine in comparison with control group, displayed a drastically decreasing in parasite burden, in their blood and tissues (P=0.000). These results indicated that ESAs with reduction of parasite load in different tissues of host could be evaluable candidate for the development of immunization strategies against toxoplasmosis.

摘要

刚地弓形虫排泄分泌抗原(ESAs)是弓形虫病免疫的候选抗原之一。为评估免疫效果,我们测定了用ESAs免疫的小鼠不同组织中弓形虫的分布动力学及寄生虫负荷。在本实验研究中,病例组(n = 18)和对照组(n = 18)的36只小鼠分别用ESAs和磷酸盐缓冲液(PBS)进行免疫。2周后,小鼠腹腔注射强毒RH株弓形虫进行攻毒。攻毒后每天(攻毒后1、2、3天及死亡前最后一天)采集血液和不同组织(脑、脾、肝、心、肾和肌肉)。使用针对B1基因的实时定量聚合酶链反应(Real time QPCR)计算寄生虫负荷。ESAs作为疫苗在不同组织中显示出不同的效果。然而,与对照组相比,接受疫苗的感染小鼠血液和组织中的寄生虫负荷急剧下降(P = 0.000)。这些结果表明,能降低宿主不同组织中寄生虫负荷的ESAs可能是开发弓形虫病免疫策略的可评估候选抗原。

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