Induction of a cellular immune response to a defined T-cell epitope as an insert in the flagellin of a live vaccine strain of Salmonella.
Induction of a cellular immune response to a defined T-cell epitope as an insert in the flagellin of a live vaccine strain of Salmonella.
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DOI：
10.1016/0264-410x(95)93308-v
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发表时间：
1995
期刊：
影响因子：
5.5
通讯作者：
N. Verma;H. Ziegler;B. Stocker;G. Schoolnik
中科院分区：
文献类型：
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作者：
N. Verma;H. Ziegler;B. Stocker;G. Schoolnik
沙门氏菌的减毒菌株已被用作疫苗来递送异源抗原，主要用于产生体液免疫应答。然而，很少有人知道他们的能力，诱导细胞介导的免疫反应的T细胞表位的另一种传染性病原体或如何最佳地提供这些表位到宿主免疫系统。为了研究这一问题，从蛾细胞色素C（MCC）的MHC II类限制性表位（残基88-103）被插入到中央高变区的鞭毛的减毒株都柏林沙门氏菌。将所得鞭毛蛋白输出到细菌表面并聚合成含有多个MCC表位拷贝的鞭毛丝。当侧接Lys-Lys组织蛋白酶B切割位点以促进其在抗原呈递细胞的内体区室内的蛋白水解释放时，MCC嵌合鞭毛蛋白表位在体外被小鼠腹腔巨噬细胞有效加工并呈递给2B 4 T-杂交瘤细胞（对MCC表位88-103具有特异性）。通过将嵌合鞭毛蛋白基因整合到疫苗株的染色体上，在H-2k小鼠中获得了表位的稳定表达和更高的免疫应答。具有MCC嵌合鞭毛蛋白（从稳定的染色体基因座表达并侧翼有组织蛋白酶B切割位点）的细菌从具有对MCC表位特异性的T细胞受体（TCR）α和β链的转基因小鼠的肝脏和脾脏中清除的速度比缺乏该表位的细菌更快。通过减毒细菌疫苗载体表达为嵌合蛋白的II类限制性表位的抗原加工和呈递可通过表位每侧上的内体蛋白酶切割位点的存在和编码序列的染色体整合来促进。
Attenuated strains of Salmonella have been used as vaccines to deliver heterologous antigens mainly to generate a humoral immune response. However, little is known about their ability to induce a cell-mediated immune response to the T-cell epitopes of another infectious agent or how optimally to deliver these epitopes to the host immune system. In order to study this question, a well defined MHC class II-restricted epitope (residues 88–103) from moth cytochrome C (MCC) was inserted into the central hypervariable domain of the flagellin of an attenuated strain of Salmonella dublin. The resulting flagellin was exported to the bacterial surface and polymerized into flagellar filaments that contained multiple copies of the MCC epitope. When flanked by Lys-Lys cathepsin B cleavage sites to facilitate its proteolytic release within the endosomal compartment of antigen-presenting cells, the MCC-chimeric flagellin epitope was efficiently processed in vitro by mouse peritoneal macrophages and presented to 2B4 T-hybridoma cells (specific for the MCC epitope 88–103). Stable expression of the epitope and a higher immune response was obtained in H-2kmice by integrating the chimeric flagellin gene into the chromosome of the vaccine strain. Bacteria with MCC-chimeric flagellins that were expressed from a stable chromosomal locus and flanked by cathepsin B cleavage sites were cleared more rapidly from the livers and spleens of transgenic mice with T-cell receptor (TCR) α and β chains specific for the MCC epitope than were bacteria lacking the epitope. Antigen processing and presentation of class II-restricted epitopes expressed as chimeric proteins by attenuated bacterial vaccine vectors may be facilitated by the presence of endosomal protease cleavage sites on each side of the epitope and by chromosomal intergration of the coding sequence.