Membrane vesicles of Clostridium perfringens type A strains induce innate and adaptive immunity.
Membrane vesicles of Clostridium perfringens type A strains induce innate and adaptive immunity.
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DOI：
10.1016/j.ijmm.2014.02.006
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发表时间：
2014-05
期刊：
影响因子：
--
通讯作者：
Curtiss R 3rd
中科院分区：
文献类型：
--
作者：
Jiang Y;Kong Q;Roland KL;Curtiss R 3rd
细菌的囊泡脱落是大多数革兰氏阴性菌和少数革兰氏阳性菌的普遍过程。本研究从革兰氏阳性病原菌产气荚膜梭菌（Clostridium perfringens，C。产气荚膜杆菌）。我们证明了囊泡生产在各种有毒和无毒A型菌株。通过对特异性抗体的阴性反应和LC-MS/MS鉴定的特异性蛋白质的缺失证明MV不含α-毒素和NetB毒素。PCR结果表明，产气荚膜菌MV含有16 S rRNA基因、α毒素基因和产气荚膜菌溶素O基因。通过一维凝胶分离和LC-MS/MS分析，我们还鉴定了囊泡中总共431个蛋白质。体外研究表明，囊泡可以内化到鼠巨噬细胞RAW264.7细胞中，而没有直接的细胞毒性作用，引起炎症细胞因子的释放，包括粒细胞集落刺激因子（G-CSF）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1（IL-1），其也可以在通过腹膜内（i. p.）路线C.产气荚膜梭菌MVs产生高滴度的IgG抗体，尤其是IgG 1。产气荚膜杆菌膜蛋白。然而，这种抗体不能提供保护，在攻击后的小鼠，但它可以稍微推迟死亡时间。我们的研究结果表明，从C。产气荚膜梭菌可能提供了一种以前未知的诱导炎症细胞因子特别是TNF-α释放的机制，这些发现可能有助于更好地理解产气荚膜梭菌的发病机制。产气荚膜杆菌感染
Vesicle shedding from bacteria is a universal process in most Gram-negative bacteria and a few Gram-positive bacteria. In this report, we isolate extracellular membrane vesicles (MVs) from the supernatants of Gram-positive pathogen Clostridium perfringens (C. perfringens). We demonstrated vesicle production in a variety of virulent and nonvirulent type A strains. MVs did not contain alpha-toxin and NetB toxin demonstrated by negative reaction to specific antibody and absence of specific proteins identified by LC-MS/MS. C. perfringens MVs contained DNA components such as 16S ribosomal RNA gene (16S rRNA), alpha-toxin gene (plc) and the perfringolysin O gene (pfoA) demonstrated by PCR. We also identified a total of 431 proteins in vesicles by 1-D gel separation and LC-MS/MS analysis. In vitro studies demonstrated that vesicles could be internalized into murine macrophage RAW264.7 cells without direct cytotoxicity effects, causing release of inflammation cytokines including granulocyte colony stimulating factor (G-CSF), tumor necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1), which could also be detected in mice injected with MVs through intraperitoneal (i.p.) route. Mice immunized with C. perfringens MVs produced high titer IgG, especially IgG1, antibodies against C. perfringens membrane proteins. However, this kind of antibody could not provide protection in mice following challenge, though it could slightly postpone the time of death. Our results indicate that release of MVs from C. perfringens could provide a previously unknown mechanism to induce release of inflammatory cytokines, especially TNF-α, these findings may contribute to a better understanding of the pathogenesis of C. perfringens infection.
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