Inhibition of caspase-1 or gasdermin-D enable caspase-8 activation in the Naip5/NLRC4/ASC inflammasome

Author(s)

Mascarenhas, Danielle P. A. ; Cerqueira, Daiane M. ; Pereira, Marcelo S. F. ; Castanheira, Fernanda V. S. ; Fernandes, Talita D. ; Manin, Graziele Z. ; Cunha, Larissa D. ; Zamboni, Dario S.

• Document type: Journal article
• Source: PLOS PATHOGENS; v. 13, n. 8 AUG 2017.
• Web of Science Citations: 33

Abstract

Legionella pneumophila is a Gram-negative, flagellated bacterium that survives in phagocytes and causes Legionnaires' disease. Upon infection of mammalian macrophages, cytosolic flagellin triggers the activation of Naip/NLRC4 inflammasome, which culminates in pyroptosis and restriction of bacterial replication. Although NLRC4 and caspase-1 participate in the same inflammasome, Nlrc4(-/-) mice and their macrophages are more permissive to L. pneumophila replication compared with Casp1/11(-/-). This feature supports the existence of a pathway that is NLRC4-dependent and caspase-1/11-independent. Here, we demonstrate that caspase-8 is recruited to the Naip5/NLRC4/ASC inflammasome in response to flagellin-positive bacteria. Accordingly, caspase-8 is activated in Casp1/11(-/-) macrophages in a process dependent on flagellin, Naip5, NLRC4 and ASC. Silencing caspase-8 in Casp1/11(-/-) cells culminated in macrophages that were as susceptible as Nlrc4(-/-) for the restriction of L. pneumophila replication. Accordingly, macrophages and mice deficient in Asc/Casp1/11(-/-) were more susceptible than Casp1/11(-/-) and as susceptible as Nlrc4(-/-) for the restriction of infection. Mechanistically, we found that caspase-8 activation triggers gasdermin-D-independent pore formation and cell death. Interestingly, caspase-8 is recruited to the Naip5/NLRC4/ASC inflammasome in wild-type macrophages, but it is only activated when caspase-1 or gasdermin-D is inhibited. Our data suggest that caspase-8 activation in the Naip5/NLRC4/ASC inflammasome enable induction of cell death when caspase- 1 or gasdermin-D is suppressed. (AU)

• FAPESP's process: 13/08216-2 - CRID - Center for Research in Inflammatory Diseases
• Grantee: Fernando de Queiroz Cunha
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 14/04684-4 - The inflammasome in the host response against intracellular pathogens and the microbial mechanisms for its evasion
• Grantee: Dario Simões Zamboni
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/09363-6 - Molecular pathogenesis and subversion of host responses in infections with Legionella spp.
• Grantee: Dario Simões Zamboni
• Support Opportunities: Regular Research Grants