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Methods for Detection of Pyrin Inflammasome Assembly in Macrophages Infected with Yersinia spp

Natasha P Medici1,2, James B Bliska3,4

  • 1Department of Molecular Genetics and Microbiology, Center for Infectious Diseases, Stony Brook University, Stony Brook, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 10, 2019
PubMed

Insights

Yersinia bacteria use effector proteins YopE and YopT to inactivate host Rho GTPases. This study details measuring pyrin inflammasome assembly, a host defense response, in macrophages infected with Yersinia lacking the YopM effector.

Area of Science:

  • Microbiology
  • Immunology
  • Cell Biology

Background:

  • Yersinia effector proteins YopE and YopT disrupt host cell signaling by inactivating Rho GTPases.
  • The host cell mounts a pro-inflammatory response, pyrin inflammasome assembly, to counteract Yersinia infection.
  • Yersinia effector YopM typically inhibits pyrin inflammasome assembly, allowing bacterial survival.

Purpose of the Study:

  • To establish a method for measuring pyrin inflammasome assembly in response to YopE and YopT.
  • To investigate host innate immune responses during Yersinia infection when YopM is absent.

Main Methods:

  • Infection of primary mouse macrophages with Yersinia strains lacking the YopM effector.
  • Detection of pyrin inflammasome assembly through downstream indicators.
  • Measurement of pyrin dephosphorylation, caspase-1 activation, IL-1β release, and pyroptosis.

Main Results:

  • YopE and YopT activities trigger pyrin inflammasome assembly in macrophages infected with yopM mutant Yersinia.
  • The assembly is characterized by pyrin dephosphorylation and caspase-1 activation.
  • Downstream consequences include IL-1β secretion and pyroptosis.

Conclusions:

  • Yersinia effector proteins YopE and YopT can induce pyrin inflammasome assembly and pyroptosis in host macrophages.
  • The absence of YopM is critical for observing this host immune response.
  • This provides a model to study Yersinia-mediated immune evasion and host defense mechanisms.
Keywords:
EffectorsInflammasomeInflammationPyrinRho GTPasesToxinsYersinia

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