Amer Amal, Franchi Luigi, Kanneganti Thirumala-Devi, Body-Malapel Mathilde, Ozören Nesrin, Brady Graham, Meshinchi Sasha, Jagirdar Rajesh, Gewirtz Andrew, Akira Shizuo, Núñez Gabriel
Department of Pathology and Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
J Biol Chem. 2006 Nov 17;281(46):35217-23. doi: 10.1074/jbc.M604933200. Epub 2006 Sep 19.
Legionella pneumophila is an intracellular bacterium that causes an acute form of pneumonia called Legionnaires' disease. After infection of human macrophages, the Legionella-containing phagosome (LCP) avoids fusion with the lysosome allowing intracellular replication of the bacterium. In macrophages derived from most mouse strains, the LCP is delivered to the lysosome resulting in Legionella degradation and restricted bacterial growth. Mouse macrophages lacking the NLR protein Ipaf or its downstream effector caspase-1 are permissive to intracellular Legionella replication. However, the mechanism by which Ipaf restricts Legionella replication is not well understood. Here we demonstrate that the presence of flagellin and a competent type IV secretion system are critical for Legionella to activate caspase-1 in macrophages. Activation of caspase-1 in response to Legionella infection also required host Ipaf, but not TLR5. In the absence of Ipaf or caspase-1 activation, the LCP acquired endoplasmic reticulum-derived vesicles, avoided fusion with the lysosome, and allowed Legionella replication. Accordingly a Legionella mutant lacking flagellin did not activate caspase-1, avoided degradation, and replicated in wild-type macrophages. The regulation of phagosome maturation by Ipaf occurred within 2 h after infection and was independent of macrophage cell death. In vivo studies confirmed that flagellin and Ipaf play an important role in the control of Legionella clearance. These results reveal that Ipaf restricts Legionella replication through the regulation of phagosome maturation, providing a novel function for NLR proteins in host defense against an intracellular bacterium.
嗜肺军团菌是一种胞内细菌,可引起一种名为军团病的急性肺炎。感染人类巨噬细胞后,含军团菌的吞噬体(LCP)避免与溶酶体融合,从而使细菌能够在细胞内复制。在大多数小鼠品系来源的巨噬细胞中,LCP会被递送至溶酶体,导致军团菌降解并限制细菌生长。缺乏NLR蛋白Ipaf或其下游效应分子caspase-1的小鼠巨噬细胞允许军团菌在细胞内复制。然而,Ipaf限制军团菌复制的机制尚不清楚。在此我们证明,鞭毛蛋白的存在和一个功能正常的IV型分泌系统对于军团菌在巨噬细胞中激活caspase-1至关重要。响应军团菌感染而激活caspase-1也需要宿主Ipaf,但不需要TLR5。在缺乏Ipaf或caspase-1激活的情况下,LCP获得了源自内质网的囊泡,避免与溶酶体融合,并允许军团菌复制。因此,一个缺乏鞭毛蛋白的军团菌突变体不会激活caspase-1,避免被降解,并在野生型巨噬细胞中复制。Ipaf对吞噬体成熟的调节发生在感染后2小时内,且与巨噬细胞死亡无关。体内研究证实,鞭毛蛋白和Ipaf在控制军团菌清除中起重要作用。这些结果表明,Ipaf通过调节吞噬体成熟来限制军团菌复制,为NLR蛋白在宿主抵御胞内细菌的防御中提供了一种新功能。
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