Analysis of Leishmania chagasi by 2-D Difference Gel Eletrophoresis (2-D DIGE) and Immunoproteomic: Identification of Novel Candidate Antigens for Diagnostic Tests and Vaccine
Analysis of Leishmania chagasi by 2-D Difference Gel Eletrophoresis (2-D DIGE) and Immunoproteomic: Identification of Novel Candidate Antigens for Diagnostic Tests and Vaccine
复制标题
DOI：
10.1021/pr101286y
复制
发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Gazzinelli, Ricardo T.
中科院分区：
文献类型：
--
作者：
Costa, Miriam M.;Andrade, Helida M.;Gazzinelli, Ricardo T.
新抗原的鉴定对于开发新的诊断测试和疫苗至关重要。我们使用DIGE比较了查斯利什曼原虫无鞭毛体和前鞭毛体形式的蛋白质表达。观察到900个无鞭毛体和前鞭毛体斑点。5无鞭毛体特异性，25 promastigote特异性，和10个蛋白质共享的两个寄生虫阶段进行了鉴定。此外，41个蛋白质鉴定在Western印迹采用2-DE和感染的狗的血清。从这些蛋白中，3和38分别与IgM和总IgG反应。总IgG识别的蛋白质在其识别IgG 1和/或IgG 2同种型方面呈现不同的模式。Western blot筛选出的所有蛋白质均定位于B细胞表位。将180个肽提交SPOT合成和免疫测定。共25个肽显示出对内脏利什曼病的血清学诊断感兴趣。此外，本研究中鉴定的所有蛋白质通过使用NetCTL软件针对T细胞表位作图，并选择用于疫苗开发的候选物。因此，对L.进行chagasi蛋白质组以鉴定新的B和T细胞表位，其具有开发诊断测试和疫苗的潜在用途。
Identification of novel antigens is essential for developing new diagnostic tests and vaccines. We used DIGE to compare protein expression in amastigote and promastigote forms of Leishmania chagasi. Nine hundred amastigote and promastigote spots were visualized. Five amastigote-specific, 25 promastigote-specific, and 10 proteins shared by the two parasite stages were identified. Furthermore, 41 proteins were identified in the Western blot employing 2-DE and sera from infected dogs. From these proteins, 3 and 38 were reactive with IgM and total IgG, respectively. The proteins recognized by total IgG presented different patterns in terms of their recognition by IgG1 and/or IgG2 isotypes. All the proteins selected by Western blot were mapped for B-cell epitopes. One hundred and eighty peptides were submitted to SPOT synthesis and immunoassay. A total of 25 peptides were shown of interest for serodiagnosis to visceral leishmaniasis. In addition, all proteins identified in this study were mapped for T cell epitopes by using the NetCTL software, and candidates for vaccine development were selected. Therefore, a large-scale screening of L. chagasi proteome was performed to identify new B and T cell epitopes with potential use for developing diagnostic tests and vaccines.