Laboratório de Pesquisas em Malária, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2013 Jun;108(4):523-8. doi: 10.1590/S0074-02762013000400022.
The genetic diversity displayed by Plasmodium falciparum, the most deadly Plasmodium species, is a significant obstacle for effective malaria vaccine development. In this study, we identified genetic polymorphisms in P. falciparum glutamate-rich protein (GLURP), which is currently being tested in clinical trials as a malaria vaccine candidate, from isolates found circulating in the Brazilian Amazon at variable transmission levels. The study was performed using samples collected in 1993 and 2008 from rural villages situated near Porto Velho, in the state of Rondônia. DNA was extracted from 126 P. falciparum-positive thick blood smears using the phenol-chloroform method and subjected to a nested polymerase chain reaction protocol with specific primers against two immunodominant regions of GLURP, R0 and R2. Only one R0 fragment and four variants of the R2 fragment were detected. No differences were observed between the two time points with regard to the frequencies of the fragment variants. Mixed infections were uncommon. Our results demonstrate conservation of GLURP-R0 and limited polymorphic variation of GLURP-R2 in P. falciparum isolates from individuals living in Porto Velho. This is an important finding, as genetic polymorphisms in B and T-cell epitopes could have implications for the immunological properties of the antigen.
恶性疟原虫(Plasmodium falciparum)是最致命的疟原虫之一,其表现出的遗传多样性是有效疟疾疫苗开发的重大障碍。在这项研究中,我们鉴定了在巴西亚马逊地区不同传播水平的循环疟原虫分离株中,目前正在临床试验中作为疟疾疫苗候选物进行测试的谷氨酸丰富蛋白(GLURP)的遗传多态性。该研究使用 1993 年和 2008 年从朗多尼亚州波多韦柳市附近农村村庄采集的样本进行。使用酚-氯仿法从 126 份疟原虫阳性厚血涂片提取 DNA,并使用针对 GLURP 的两个免疫显性区域(R0 和 R2)的特异性引物进行巢式聚合酶链反应。仅检测到一个 R0 片段和 R2 片段的四个变体。两个时间点之间的片段变体频率没有差异。混合感染并不常见。我们的结果表明,来自波多韦柳市居民的疟原虫分离株中 GLURP-R0 保持保守,GLURP-R2 的多态性有限。这是一个重要的发现,因为 B 和 T 细胞表位的遗传多态性可能会影响抗原的免疫特性。