It is conceivable that other inflammasomes are also inhibited by the drug (see below). 

Lamkanfi M, Mueller JL, Vitari AC, Misaghi S, Fedorova A, Deshayes K, Lee WP,

Hoffman HM, Dixit VM. Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. J Cell

Biol. 2009 Oct 5;187(1):61-70.

What is your inflammasome stimulus?

Hi Jordan,

The cells are challanged with mycobacteria.

Hi - what concentration are you using glyburide at? Are you controlling for toxicity?

Hi Rebecca, 

Its at 50micromolar which isn't causing much cell death.

Hi Mary,

At that concentration I haven't seen any effects on AIM2 or NLRC4 activation but that was using macrophages so there may be some differences with monocytes. In the Lamkanfi/Dixit paper in Fig 3. F they show that glyburide blocks caspase-1 activation in response to LPS transfection with DOTAP which we now know activates the non-canonical caspase-11 inflammasome. In murine macrophages IL-1b production in response to intracellular LPS is both NLRP3 and caspase-11 dependent but cell death is completely caspase-11 dependent (it's in the supplemental figs in this paper http://www.ncbi.nlm.nih.gov/pubmed/23887873). So if your mycobacteria could access the cytosol they may activate caspase-11 (or caspase 4/5 in human cells) and this could be blocked by glyburide. What readouts are you using? Are there any differences between IL-1b production vs cell death caused by mycobacteria challenge?

This might be more confusing than helpful!

Hi Rebecca,

Thanks for the information about the AIM2 and NLRC4 inflammasomes. We are seeing inhibition of  IL-1b secretion and of cell death  by glyburide. We haven't checked yet to see whether siRNA knockdown of NLRP3 inhibits IL-1b production but it has no effect on cell death.  Knockdown of caspase 1 blocks cell death to a very small extent so it might be worth looking at caspase 4/5 as well as you suggest.

Hi Mary,

Have you phenotypically confirmed your siRNA knockdown of NLRP3 against a canonical activator such as nigericin or ATP for reduction of IL1b secretion and cell death?

Not yet Jordan. We should definitely check that.