Delivery of biologically, active anti-inflammatory cytokines IL-10 and IL-1ra in vivo by the shigella type III secretion apparatus
Delivery of biologically, active anti-inflammatory cytokines IL-10 and IL-1ra in vivo by the shigella type III secretion apparatus
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DOI：
10.4049/jimmunol.180.6.4292
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发表时间：
2008-03-15
影响因子：
4.4
通讯作者：
Allaoui, Abdehnounaaim
中科院分区：
文献类型：
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作者：
Chamekh, Mustapha;Phalipon, Armelle;Allaoui, Abdehnounaaim
许多革兰氏阴性细菌的致病性依赖于III型分泌（T3 S）装置，其用于将细菌效应物递送到宿主细胞质中，从而允许细菌操纵宿主细胞细胞骨架网络以及干扰细胞内信号传导途径。在这项研究中，我们研究了福氏志贺菌T3 SA作为生物活性分子如细胞因子的体内递送系统的潜力。将抗炎细胞因子IL-10和IL-1受体拮抗剂（IL-1 ra）与志贺氏菌T3 S效应子IpaH9.8的前30或60个残基或小肠结肠炎耶尔森菌效应子YopE的前50个残基基因融合，并在S.弗莱克斯内里。YopE（50）-IL-10、IpaH（60）-IL-10和IpaH（60）-IL-1 ra通过志贺菌的T3 S装置有效分泌。此外，这些重组蛋白在体外不损害细菌的侵袭能力。在鼠模型中，表达YopE（50）-IL-10、IpaH（60）-IL-10和IpaH（60）-IL-1 ra的志贺氏菌菌株在小鼠中诱导了较低的死亡率，这与减少的炎症和与野生型志贺氏菌相比细菌在肺组织内的受限定位有关。此外，TNF-α和IL-1 β mRNA的水平降低，在肺部感染后的IL-10-和IL-1 ra分泌志贺氏菌，分别。这些发现表明志贺氏菌T3 S装置可以在体内递送生物活性细胞因子，从而为使用减毒细菌递送用于免疫调节或基因治疗目的的蛋白质开辟了新的途径。
Pathogenicity of many Gram-negative bacteria relies on a type III secretion (T3S) apparatus, which is used for delivery of bacterial effectors into the host cell cytoplasm allowing the bacteria to manipulate host cell cytoskeleton network as well as to interfere with intracellular signaling pathways. In this study, we investigated the potential of the Shigella flexneri T3SA as an in vivo delivery system for biologically active molecules such as cytokines. The anti-inflammatory cytokines IL-10 and IL-I receptor antagonist (IL-Ira) were genetically fused to the first 30 or 60 residues of the Shigella T3S effector IpaH9.8 or to the first 50 residues of the Yersinia enterocolitica effector YopE and the recombinant fusion proteins were expressed in S. flexneri. YopE(50)-IL-10, lpaH(60)-IL-10, and IpaH(60)-IL-1ra were efficiently secreted via the T3S apparatus of Shigella. Moreover, these recombinant proteins did not impair the invasive ability of the bacteria in vitro. In a murine model, Shigella strains expressing YopE(50)-IL-10, lpaH(60)-IL-10, and lpaH(60)-IL-lra induced a lower mortality in mice that was associated with reduced inflammation and a restricted localization of bacteria within the lung tissues as compared with wild-type Shigella. Moreover, the level of TNF-alpha and IL-1 beta mRNA were reduced in the lungs following infection by IL-10- and IL-1ra-secreting Shigella, respectively. These findings demonstrate that the Shigella T3S apparatus can deliver biologically active cytokines in vivo, thus opening new avenues for the use of attenuated bacteria to deliver proteins for immunomodulation or gene therapy purposes.