Vance et al. provide genetic proof for the specificity and essentiality of NAIP proteins for inflammasome responses to specific bacterial ligands in vivo.

Inflammasomes are cytosolic multi-protein complexes that detect infection or cellular damage and activate the Caspase-1 (CASP1) protease. The NAIP5/NLRC4 inflammasome detects bacterial flagellin and is essential for resistance to the flagellated intracellular bacterium Legionella pneumophila . The effectors required downstream of NAIP5/NLRC4 to restrict bacterial replication remain unclear. Upon NAIP5/NLRC4 activation, CASP1 cleaves and activates the pore-forming protein Gasdermin-D (GSDMD) and the effector caspase-7 (CASP7). However, Casp1 –/– (and Casp1/11 –/– ) mice are only partially susceptible to L . pneumophila and do not phenocopy Nlrc4 –/– mice, because NAIP5/NLRC4 also activates CASP8 for restriction of L . pneumophila infection. Here we show that CASP8 promotes the activation of CASP7 and that Casp7/1/11 –/– and Casp8/1/11 –/– mice recapitulate the full susceptibility of Nlrc4 –/– mice. Gsdmd –/– mice exhibit only mild susceptibility to L . pneumophila , but Gsdmd –/– Casp7 –/– mice are as susceptible as the Nlrc4 –/– mice. These results demonstrate that GSDMD and CASP7 are the key substrates downstream of NAIP5/NLRC4/CASP1/8 required for resistance to L . pneumophila .

Inflammasomes are cytosolic multiprotein complexes that initiate protective immunity in response to infection, and can also drive auto-inflammatory diseases, but the cell types and signalling pathways that cause these diseases remain poorly understood. Inflammasomes are broadly expressed in haematopoietic and non-haematopoietic cells and can trigger numerous downstream responses including production of IL-1β, IL-18, eicosanoids and pyroptotic cell death. Here we show a mouse model with endogenous NLRC4 inflammasome activation in Lysozyme2 + cells (monocytes, macrophages and neutrophils) in vivo exhibits a severe systemic inflammatory disease, reminiscent of human patients that carry mutant auto-active NLRC4 alleles. Interestingly, specific NLRC4 activation in Mrp8 + cells (primarily neutrophil lineage) is sufficient to cause severe inflammatory disease. Disease is ameliorated on an Asc −/− background, and can be suppressed by injections of anti-IL-1 receptor antibody. Our results provide insight into the mechanisms by which NLRC4 inflammasome activation mediates auto-inflammatory disease in vivo.

The bat fly (Trichobius major) is a blood-feeding ectoparasite of the cave myotis (Myotis velifer). A recent mitochondrial DNA (mtDNA) study examining population structure of T. major in the South Central United States detected a single haplotype from all individuals examined (N = 48 from 12 different caves), representing one of only a few known examples of such widespread mtDNA uniformity. We examined nuclear genetic diversity using amplified fragment length polymorphism and detected high levels of nuclear genetic diversity in all populations sampled. Amplified fragment length polymorphism analyses indicated significant levels of gene flow among caves >700 km apart, suggesting the absence of mtDNA diversity in T. major is the result of a selective sweep, not a demographic event (i.e., a recent bottleneck). One mechanism by which mtDNA sweeps occur in arthropods is through bacterial parasites that manipulate host reproduction and mtDNA inheritance. We used PCR to test for the presence of all known reproductive parasites and detected a widespread infection (91.33% infection rate) of T. major with a novel Arsenophonus bacterium, as well as the infection of 2 individuals (1.16% infection rate) with a novel strain of Rickettsia. We discuss the implications for T. major phylogeography and the necessity of a bigenomic approach in arthropod population genetics.