Laboratório de Biologia Molecular de Parasitas e Vetores, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Rio de Janeiro, Brazil.
PLoS One. 2010 Mar 22;5(3):e9795. doi: 10.1371/journal.pone.0009795.
Malaria affects 300 million people worldwide every year and is endemic in 22 countries in the Americas where transmission occurs mainly in the Amazon Region. Most malaria cases in the Americas are caused by Plasmodium vivax, a parasite that is almost impossible to cultivate in vitro, and Anopheles aquasalis is an important malaria vector. Understanding the interactions between this vector and its parasite will provide important information for development of disease control strategies. To this end, we performed mRNA subtraction experiments using A. aquasalis 2 and 24 hours after feeding on blood and blood from malaria patients infected with P. vivax to identify changes in the mosquito vector gene induction that could be important during the initial steps of infection. A total of 2,138 clones of differentially expressed genes were sequenced and 496 high quality unique sequences were obtained. Annotation revealed 36% of sequences unrelated to genes in any database, suggesting that they were specific to A. aquasalis. A high number of sequences (59%) with no matches in any databases were found 24 h after infection. Genes related to embryogenesis were down-regulated in insects infected by P. vivax. Only a handful of genes related to immune responses were detected in our subtraction experiment. This apparent weak immune response of A. aquasalis to P. vivax infection could be related to the susceptibility of this vector to this important human malaria parasite. Analysis of some genes by real time PCR corroborated and expanded the subtraction results. Taken together, these data provide important new information about this poorly studied American malaria vector by revealing differences between the responses of A. aquasalis to P. vivax infection, in relation to better studied mosquito-Plasmodium pairs. These differences may be important for the development of malaria transmission-blocking strategies in the Americas.
疟疾每年影响全球 3 亿人,在美洲 22 个国家流行,这些国家的传播主要发生在亚马逊地区。美洲的大多数疟疾病例由疟原虫引起,这种寄生虫几乎不可能在体外培养,而按蚊aquasalis 是一种重要的疟疾媒介。了解这种媒介及其寄生虫之间的相互作用将为开发疾病控制策略提供重要信息。为此,我们使用 A. aquasalis 2 和 24 小时后喂食血液和感染疟原虫 vivax 的疟疾病人血液进行 mRNA 消减实验,以鉴定蚊子载体基因诱导的变化,这些变化在感染的初始步骤中可能很重要。总共对 2138 个差异表达基因的克隆进行了测序,获得了 496 个高质量的独特序列。注释显示 36%的序列与任何数据库中的基因无关,表明它们是 A. aquasalis 特有的。感染后 24 小时发现大量(59%)在任何数据库中都没有匹配的序列。与疟原虫感染有关的胚胎发生相关基因下调。在我们的消减实验中仅检测到少数与免疫反应相关的基因。A. aquasalis 对疟原虫感染的这种明显较弱的免疫反应可能与该载体对这种重要的人类疟疾寄生虫的易感性有关。通过实时 PCR 对一些基因进行分析,证实并扩展了消减实验的结果。总的来说,这些数据通过揭示 A. aquasalis 对疟原虫感染的反应与研究较好的蚊子-疟原虫对之间的差异,为研究这种研究较少的美洲疟疾媒介提供了重要的新信息。这些差异可能对在美洲开发阻断疟疾传播的策略很重要。