Combinational deletion of three membrane protein-encoding genes highly attenuates yersinia pestis while retaining immunogenicity in a mouse model of pneumonic plague.
Combinational deletion of three membrane protein-encoding genes highly attenuates yersinia pestis while retaining immunogenicity in a mouse model of pneumonic plague.
复制标题
三个膜蛋白编码基因的组合删除可高度减弱鼠疫耶尔森氏菌，同时保留肺鼠疫小鼠模型中的免疫原性。
DOI：
10.1128/iai.02778-14
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发表时间：
2015
影响因子：
3.1
通讯作者：
Chopra,AshokK
中科院分区：
文献类型：
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作者：
Tiner,BethanyL;Sha,Jian;Kirtley,MichelleL;Erova,TatianaE;Popov,VsevolodL;Baze,WallaceB;vanLier,ChristinaJ;Ponnusamy,Duraisamy;Andersson,JourdanA;Motin,VladimirL;Chauhan,Sadhana;Chopra,AshokK
以前，我们表明，编码布劳恩脂蛋白（Lpp）和MsbB基因的缺失减毒鼠疫耶尔森氏菌CO92在小鼠和大鼠模型的腺鼠疫和肺鼠疫。虽然Lpp激活Toll样受体2，但MsbB酰基转移酶修饰脂多糖。在此，我们从野生型（WT）菌株CO92或其slpp单突变体和Δ lpp Δ msbB双突变体中删除了该基因（编码附着-侵袭位点）。虽然在肺鼠疫小鼠模型中，Δ ailsingle突变体与WT细菌相比减毒程度最低，但Δ lpp Δ aildouble突变体和Δ lpp Δ msbB Δ ailtriple突变体的减毒程度逐渐增加，后者在相当于6，800 LD50的WT CO92的50%致死剂量（LD50）下无法杀死小鼠。根据抗体同种型，感染mu的动物产生了平衡的基于TH1和TH2的免疫应答。三重突变体从小鼠器官中迅速清除，同时减少各种细胞因子的产生和组织病理学病变。当用增加剂量的三重突变体感染的存活动物随后在第24天用生物发光WT CO92菌株（20至28 LD 50）攻击时，40至70%的小鼠存活，并有效清除了入侵的病原体，如通过体内成像在真实的时间中所观察到的。与WT CO92相比，三重突变体从动物中的快速清除与人源性HeLa和A549肺泡上皮细胞的粘附和侵袭降低以及其不能在这些细胞以及MH-S小鼠肺泡和原代人巨噬细胞中细胞内存活有关。与WT CO92相比，三重突变体引起的巨噬细胞中细胞因子产生的早期爆发以及突变体对人血清杀菌作用的敏感性将进一步增加细菌清除。总之，从Δ lpp Δ msbB双突变体中缺失该基因严重减弱了Y。鼠疫菌CO92在小鼠模型中诱发肺鼠疫，同时保留随后针对感染的保护所需的免疫原性。
Previously, we showed that deletion of genes encoding Braun lipoprotein (Lpp) and MsbB attenuated Yersinia pestis CO92 in mouse and rat models of bubonic and pneumonic plague. While Lpp activates Toll-like receptor 2, the MsbB acyltransferase modifies lipopolysaccharide. Here, we deleted theailgene (encoding theattachment-invasionlocus) from wild-type (WT) strain CO92 or itslppsingle and ΔlppΔmsbBdouble mutants. While the Δailsingle mutant was minimally attenuated compared to the WT bacterium in a mouse model of pneumonic plague, the ΔlppΔaildouble mutant and the ΔlppΔmsbBΔailtriple mutant were increasingly attenuated, with the latter being unable to kill mice at a 50% lethal dose (LD50) equivalent to 6,800 LD50s of WT CO92. The mutant-infected animals developed balanced TH1- and TH2-based immune responses based on antibody isotyping. The triple mutant was cleared from mouse organs rapidly, with concurrent decreases in the production of various cytokines and histopathological lesions. When surviving animals infected with increasing doses of the triple mutant were subsequently challenged on day 24 with the bioluminescent WT CO92 strain (20 to 28 LD50s), 40 to 70% of the mice survived, with efficient clearing of the invading pathogen, as visualized in real time byin vivoimaging. The rapid clearance of the triple mutant, compared to that of WT CO92, from animals was related to the decreased adherence and invasion of human-derived HeLa and A549 alveolar epithelial cells and to its inability to survive intracellularly in these cells as well as in MH-S murine alveolar and primary human macrophages. An early burst of cytokine production in macrophages elicited by the triple mutant compared to WT CO92 and the mutant's sensitivity to the bactericidal effect of human serum would further augment bacterial clearance. Together, deletion of theailgene from the ΔlppΔmsbBdouble mutant severely attenuated Y. pestis CO92 to evoke pneumonic plague in a mouse model while retaining the required immunogenicity needed for subsequent protection against infection.