A fusogenic peptide expressed on the surface of Salmonella enterica elicits CTL responses to a dengue virus epitope
A fusogenic peptide expressed on the surface of Salmonella enterica elicits CTL responses to a dengue virus epitope
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DOI：
10.1016/j.vaccine.2007.03.047
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发表时间：
2007-06-28
期刊：
影响因子：
5.5
通讯作者：
Gonzalez-Bonilla, C. R.
中科院分区：
文献类型：
--
作者：
Luria-Perez, R.;Cedillo-Barron, L.;Gonzalez-Bonilla, C. R.
减毒沙门氏菌菌株被广泛用作抗原的活载体，因为它们引起针对乘客抗原的粘膜和全身免疫。然而，它们通常引起差的细胞毒性T细胞（CTL）应答，因为沙门氏菌驻留在空泡区室中，并且过客抗原必须行进到胞质溶胶并通过MHC 1类依赖性途径加工以模拟CTL。为了解决这个问题，我们设计了一种融合蛋白来破坏吞噬体膜的稳定性，并允许登革热表位到达细胞质。该融合蛋白通过自身转运蛋白MisL的β结构域展示在沙门氏菌鼠伤寒血清型SL3261的细菌表面。过客A结构域从N-末端起包含融合序列、来自登革病毒2型的NS3蛋白298 - 306-氨基酸CTL表位、分子标签和蛋白酶OmpT的识别位点以将其释放到环境中。使用针对分子标签的抗体通过IFA和流式细胞术证明融合蛋白在细菌表面上的展示。使用OmpT+大肠杆菌菌株通过流式细胞术证明融合蛋白-登革肽的裂解。展示融合登革肽的重组沙门氏菌菌株能够裂解红细胞，诱导特异性增殖反应，并引发CTL反应。这些结果表明，含有融合序列的重组融合蛋白提供了一个有前途的系统，诱导CTL的活载体疫苗。(C)2007爱思唯尔有限公司保留所有权利。
Attenuated Salmonella strains are used widely as live carriers of antigens because they elicit both mucosal and systemic immunity against passenger antigens. However, they generally evoke poor cytotoxic T cell (CTL) responses because Salmonella resides within vacuolar compartments and the passenger antigens must travel to the cytosol and be processed through the MHC class 1-dependent pathway to simulate CTLs. To address this problem, we designed a fusion protein to destabilize the phagosome membrane and allow a dengue epitope to reach the cytosol. The fusion protein was displayed on the bacterial surface of Salmonella enterica serovar Typhimurium SL3261 through the beta domain of the autotransporter MisL. The passenger a domain contained, from the N-terminus, a fusogenic sequence, the NS3 protein 298-306-amino acid CTL epitope from the dengue virus type 2, a molecular tag, and a recognition site for the protease OmpT to release it to the milieu. Display of the fusion protein on the bacterial surface was demonstrated by IFA and flow cytometry using antibodies against the molecular tag. Cleavage of the fusogenic protein-dengue peptide was demonstrated by flow cytometry using OmpT+ Escherichia coli strains. The recombinant Salmonella strains displaying the fusogenic-dengue peptide were able to lyse erythrocytes, induced specific proliferative responses, and elicited CTL responses. These results suggest that the recombinant fusion proteins containing fusogenic sequences provide a promising system to induce CTLs by live vector vaccines. (C) 2007 Elsevier Ltd. All rights reserved.