Panek Johan, El Alaoui Hicham, Mone Anne, Urbach Serge, Demettre Edith, Texier Catherine, Brun Christine, Zanzoni Andreas, Peyretaillade Eric, Parisot Nicolas, Lerat Emmanuelle, Peyret Pierre, Delbac Frederic, Biron David G
Clermont Université, Université Blaise Pascal, Laboratoire Microorganismes: Génome et Environnement, Clermont-Ferrand, France; CNRS, UMR 6023, LMGE, Aubière, France.
Functional Proteomics Platform. UMR CNRS 5203, Montpellier, France.
PLoS One. 2014 Jun 26;9(6):e100791. doi: 10.1371/journal.pone.0100791. eCollection 2014.
Intracellular pathogens including bacteria, viruses and protozoa hijack host cell functions to access nutrients and to bypass cellular defenses and immune responses. These strategies have been acquired through selective pressure and allowed pathogens to reach an appropriate cellular niche for their survival and growth. To get new insights on how parasites hijack host cellular functions, we developed a SILAC (Stable Isotope Labeling by Amino Acids in Cell culture) quantitative proteomics workflow. Our study focused on deciphering the cross-talk in a host-parasite association, involving human foreskin fibroblasts (HFF) and the microsporidia Anncaliia algerae, a fungus related parasite with an obligate intracellular lifestyle and a strong host dependency. The host-parasite cross-talk was analyzed at five post-infection times 1, 6, 12 and 24 hours post-infection (hpi) and 8 days post-infection (dpi). A significant up-regulation of four interferon-induced proteins with tetratricopeptide repeats IFIT1, IFIT2, IFIT3 and MX1 was observed at 8 dpi suggesting a type 1 interferon (IFN) host response. Quantitative alteration of host proteins involved in biological functions such as signaling (STAT1, Ras) and reduction of the translation activity (EIF3) confirmed a host type 1 IFN response. Interestingly, the SILAC approach also allowed the detection of 148 A. algerae proteins during the kinetics of infection. Among these proteins many are involved in parasite proliferation, and an over-representation of putative secreted effectors proteins was observed. Finally our survey also suggests that A. algerae could use a transposable element as a lure strategy to escape the host innate immune system.
包括细菌、病毒和原生动物在内的细胞内病原体劫持宿主细胞功能以获取营养，并绕过细胞防御和免疫反应。这些策略是通过选择性压力获得的，使病原体能够找到适合其生存和生长的细胞生态位。为了深入了解寄生虫如何劫持宿主细胞功能，我们开发了一种SILAC（细胞培养中氨基酸稳定同位素标记）定量蛋白质组学工作流程。我们的研究重点是破译宿主-寄生虫关联中的相互作用，涉及人包皮成纤维细胞（HFF）和微孢子虫安氏无胆虫，这是一种与真菌相关的寄生虫，具有专性细胞内生活方式且对宿主有很强的依赖性。在感染后1、6、12和24小时以及感染后8天这五个感染时间点分析宿主-寄生虫的相互作用。在感染后8天观察到四种具有四肽重复序列的干扰素诱导蛋白IFIT1、IFIT2、IFIT3和MX1显著上调，表明存在1型干扰素（IFN）宿主反应。参与信号传导（STAT1、Ras）等生物学功能的宿主蛋白的定量变化以及翻译活性（EIF3）的降低证实了宿主1型干扰素反应。有趣的是，SILAC方法还能够在感染动力学过程中检测到148种安氏无胆虫蛋白。在这些蛋白中，许多与寄生虫增殖有关，并且观察到推定的分泌效应蛋白的比例过高。最后，我们的研究还表明，安氏无胆虫可能利用转座元件作为一种诱捕策略来逃避宿主的先天免疫系统。
