de Araújo Cintia F, Silva Virgínia M G, Cronemberger-Andrade Andre, Aragão-França Luciana S, Rocha Viviane C J, Santos Priscila S L, Pontes-de-Carvalho Lain
Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua Waldemar Falcão 121, Salvador, Brazil.
BMC Res Notes. 2014 Feb 1;7:70. doi: 10.1186/1756-0500-7-70.
It has been reported that repeated intravenous injections of a relatively large amount of Leishmania amazonensis amastigote extract (LaE) in BALB/c mice exacerbates the infection of these mice by Leishmania braziliensis. The identification of the extract active principle(s) through physicochemical purification often involves dilution and losses of protein in the course of successive purification procedures. The large amount of the extract required to induce the phenomenon, therefore, hinders the carrying out of experiments aimed at identifying the active molecule(s) through extract purification. In the present work, a dose-response experiment was done to find out if smaller amounts of LaE than that necessary to be used by the intravenous route would reproduce the phenomenon when injected by the intradermal route. In addition, it was also investigated whether a Leishmania braziliensis amastigote extract (LbE) would exert the same effect and whether the effect would occur in C57BL/6 mice.
It was found that a single injection of either LaE or LbE containing 5 μg of protein was capable of enhancing the infection in BALB/c but not in C57BL/6 mice. In addition, it was observed that the largest tested doses of LbE (containing 30 and 180 μg of protein) failed to enhance the infection by L. braziliensis, whereas all doses of LaE enhanced equally that infection.
Those results indicate the possible existence in LbE, and not in LaE, of molecules that interfere with the extract infection-enhancing activity when it is injected in large amounts, and that the inoculation of Leishmania extracts through the intravenous and intradermal routes potentiate the infection by L. braziliensis through the same mechanism.
据报道,在BALB/c小鼠中反复静脉注射相对大量的亚马逊利什曼原虫无鞭毛体提取物(LaE)会加剧这些小鼠被巴西利什曼原虫的感染。通过物理化学纯化鉴定提取物的活性成分通常会在连续纯化过程中涉及蛋白质的稀释和损失。因此,诱导该现象所需的大量提取物阻碍了旨在通过提取物纯化鉴定活性分子的实验的进行。在本研究中,进行了剂量反应实验,以确定经皮内途径注射比静脉途径所需量更少的LaE是否会重现该现象。此外,还研究了巴西利什曼原虫无鞭毛体提取物(LbE)是否会产生相同的效果以及该效果是否会在C57BL/6小鼠中出现。
发现单次注射含有5μg蛋白质的LaE或LbE能够增强BALB/c小鼠的感染,但不能增强C57BL/6小鼠的感染。此外,观察到测试的最大剂量的LbE(含有30和180μg蛋白质)未能增强巴西利什曼原虫的感染,而所有剂量的LaE均同等程度地增强了该感染。
这些结果表明,当大量注射时,LbE中可能存在干扰提取物感染增强活性的分子,而LaE中不存在,并且通过静脉内和皮内途径接种利什曼原虫提取物通过相同机制增强巴西利什曼原虫的感染。