Lactobacillus plantarum displaying CCL3 chemokine in fusion with HIV-1 Gag derived antigen causes increased recruitment of T cells.
Lactobacillus plantarum displaying CCL3 chemokine in fusion with HIV-1 Gag derived antigen causes increased recruitment of T cells.
复制标题
在与HIV-1插入式抗原融合中显示CCL3趋化因子的乳杆菌溶剂因子导致T细胞的募集增加。
DOI：
10.1186/s12934-015-0360-z
复制
发表时间：
2015-10-22
影响因子：
6.4
通讯作者：
Øynebråten I
中科院分区：
文献类型：
--
作者：
Kuczkowska K;Mathiesen G;Eijsink VG;Øynebråten I
趋化因子因其招募免疫细胞的能力而成为疫苗佐剂的有吸引力的候选者。以乳酸菌(LAB)为基础的运送工具有可能被用作廉价和安全的疫苗接种选择。在LAB表面产生的趋化因子可能会潜在地增强对抗原的免疫应答，这种方法可以考虑在未来的粘膜疫苗的开发中使用。我们已经构建了植物乳杆菌菌株，它们的表面显示了一种趋化因子。植物乳杆菌经过基因工程，可以表达一种名为CCL3Gag的蛋白质，并将其锚定在表面。CCL3Gag是由截短的HIV-1Gag抗原和小鼠趋化因子CCL3(也称为MIP-1α)组成的融合蛋白。探索了不同的表面锚定策略：(1)基于脂盒的共价膜锚定，(2)由索尔特酶介导的共价细胞壁锚定，(3)基于LysM的非共价细胞壁锚定，以及(4)基于N端信号肽的跨膜锚定。Western blotting、流式细胞仪和免疫荧光显微镜证实蛋白质的产生和正确的定位。通过趋化实验，我们证明了产生CCL3Gag的植物乳杆菌菌株能够在体外募集免疫细胞。结果表明，工程化植物乳杆菌能够产生固定在细菌表面的功能性趋化蛋白。我们观察到，表面展示的CCL3Gag的活性因所使用的锚定类型而不同。作为细菌疫苗一部分的趋化因子可以增加免疫细胞的招募，从而增强免疫系统对疫苗的反应。本文的在线版本(doi：10.1186/s12934-0150360-z)包含补充材料，授权用户可以使用。
Chemokines are attractive candidates for vaccine adjuvants due to their ability to recruit the immune cells. Lactic acid bacteria (LAB)-based delivery vehicles have potential to be used as a cheap and safe option for vaccination. Chemokine produced on the surface of LAB may potentially enhance the immune response to an antigen and this approach can be considered in development of future mucosal vaccines. We have constructed strains of Lactobacillusplantarum displaying a chemokine on their surface. L. plantarum was genetically engineered to express and anchor to the surface a protein called CCL3Gag. CCL3Gag is a fusion protein comprising of truncated HIV-1 Gag antigen and the murine chemokine CCL3, also known as MIP-1α. Various surface anchoring strategies were explored: (1) a lipobox-based covalent membrane anchor, (2) sortase-mediated covalent cell wall anchoring, (3) LysM-based non-covalent cell wall anchoring, and (4) an N-terminal signal peptide-based transmembrane anchor. Protein production and correct localization were confirmed using Western blotting, flow cytometry and immunofluorescence microscopy. Using a chemotaxis assay, we demonstrated that CCL3Gag-producing L. plantarum strains are able to recruit immune cells in vitro. The results show the ability of engineered L. plantarum to produce a functional chemotactic protein immobilized on the bacterial surface. We observed that the activity of surface-displayed CCL3Gag differed depending on the type of anchor used. The chemokine which is a part of the bacteria-based vaccine may increase the recruitment of immune cells and, thereby, enhance the reaction of the immune system to the vaccine. The online version of this article (doi:10.1186/s12934-015-0360-z) contains supplementary material, which is available to authorized users.