Gourlay Louise J, Peri Claudio, Ferrer-Navarro Mario, Conchillo-Solé Oscar, Gori Alessandro, Rinchai Darawan, Thomas Rachael J, Champion Olivia L, Michell Stephen L, Kewcharoenwong Chidchamai, Nithichanon Arnone, Lassaux Patricia, Perletti Lucia, Longhi Renato, Lertmemongkolchai Ganjana, Titball Richard W, Daura Xavier, Colombo Giorgio, Bolognesi Martino
Department of Biosciences, University of Milan, 20133 Milan, Italy.
Chem Biol. 2013 Sep 19;20(9):1147-56. doi: 10.1016/j.chembiol.2013.07.010. Epub 2013 Aug 29.
We solved the crystal structure of Burkholderia pseudomallei acute phase antigen BPSL2765 in the context of a structural vaccinology study, in the area of melioidosis vaccine development. Based on the structure, we applied a recently developed method for epitope design that combines computational epitope predictions with in vitro mapping experiments and successfully identified a consensus sequence within the antigen that, when engineered as a synthetic peptide, was selectively immunorecognized to the same extent as the recombinant protein in sera from melioidosis-affected subjects. Antibodies raised against the consensus peptide were successfully tested in opsonization bacterial killing experiments and antibody-dependent agglutination tests of B. pseudomallei. Our strategy represents a step in the development of immunodiagnostics, in the production of specific antibodies and in the optimization of antigens for vaccine development, starting from structural and physicochemical principles.
在类鼻疽病疫苗研发领域的结构疫苗学研究中，我们解析了类鼻疽伯克霍尔德菌急性期抗原BPSL2765的晶体结构。基于该结构，我们应用了一种最近开发的表位设计方法，该方法将计算表位预测与体外定位实验相结合，并成功在抗原中鉴定出一个共有序列。当将该共有序列设计成合成肽时，在类鼻疽病患者血清中，它能被选择性免疫识别，且与重组蛋白的识别程度相同。针对该共有肽产生的抗体，在类鼻疽伯克霍尔德菌的调理吞噬杀菌实验和抗体依赖性凝集试验中成功通过了测试。我们的策略从结构和物理化学原理出发，代表了免疫诊断学发展、特异性抗体制备以及疫苗研发抗原优化方面的一个进展。
