Brobey Reynolds K B, Mei Fang C, Cheng Xiaodong, Soong Lynn
Department of Pathology, Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Galveston, TX 77555-1070, USA.
Braz J Infect Dis. 2006 Feb;10(1):1-6. doi: 10.1590/s1413-86702006000100001. Epub 2006 Jun 2.
The outcome of Leishmania infections is determined by both the parasite species and the host genetic makeup. While much has been learned regarding immune responses to this parasite, our knowledge on parasite-derived factors is limited. The recent completion of the L. major and L. infantum genome sequence projects and concurrent advancement in proteomics technology would greatly accelerate the search for novel Leishmania proteins. Using a proteomics-based approach to study species-specific Leishmania proteins, we developed high-resolution, broad pH (3-10) two-dimensional gel electrophoresis (2-DE) separations to determine protein-expression profiles between highly infectious forms of the parasitic species L. amazonensis (New World) and L. major (Old World). Approximately 1,650 and 1,530 distinct protein spots were detected in the L. amazonensis and L. major gels, respectively. While a vast majority of the spots had similar distribution and intensity, a few were computationally defined as preferentially expressed in L. amazonensis in comparison to L. major, or vice versa. These data attest to the feasibility of establishing a 2-DE-based protein array for inter-species profiling of Leishmania proteins and provide the framework for future design of proteome studies of Leishmania.
利什曼原虫感染的结果由寄生虫种类和宿主基因构成共同决定。尽管我们对针对这种寄生虫的免疫反应已有很多了解，但对于寄生虫衍生因子的认识却很有限。近来，硕大利什曼原虫和婴儿利什曼原虫基因组测序项目的完成以及蛋白质组学技术的同步发展，将极大地加速新型利什曼原虫蛋白的搜寻。我们采用基于蛋白质组学的方法来研究利什曼原虫的种特异性蛋白，开发了高分辨率、宽pH值（3 - 10）的二维凝胶电泳（2 - DE）分离技术，以确定寄生性物种亚马逊利什曼原虫（新大陆）和硕大利什曼原虫（旧大陆）高感染性形态之间的蛋白质表达谱。在亚马逊利什曼原虫和硕大利什曼原虫的凝胶中分别检测到约1650个和1530个不同的蛋白斑点。虽然绝大多数斑点具有相似的分布和强度，但通过计算确定，与硕大利什曼原虫相比，少数斑点在亚马逊利什曼原虫中优先表达，反之亦然。这些数据证明了建立基于二维电泳的蛋白质阵列用于利什曼原虫种间蛋白谱分析的可行性，并为未来利什曼原虫蛋白质组研究的设计提供了框架。
