Ever wondered why there is so little #microscopy of #GSDMD pores? Check out how Lisa Schiffelers used #Nanobodies to stabilize monomeric GSDMD in the plasma membrane, to dissect the steps of pore formation, and to develop some new ideas for therapy.

Late #WeekendRead: We describe a novel reporter to visualize endogenous #inflammasome assembly. Suitable for cell lines, primary cells, and tissues as well as quantification by #microscopy and #flowcytometry. Happy to share.

RT @schmidburgk: How could we identify key genes involved in any biological process and cell type? Please read on about Nuclear In-Situ Seq….

RT @MarionKoopmans: and so it begins. science needs to start getting serious about how how to deal with this type of attacks, sowing dis….

Check out Lisa‘s take on her recent paper on the mechanism (and potential therapeutic targeting) of #GSDMD pore formation using #nanobodies.

RT @FelipeOpaz_o: It is out! #NanoPlex is the universal strategy for multiplexing any immunofluorescence using erasable #nanobodies from @_….

RT @ISMMSscopes: 😯😯.

RT @DagmarWachten: Teaching the next generation of scientists all about what we do in the lab #TürenaufmitderMaus – big thank you to my who….

RT @Synth_Immune: Epic effort from Flo & first author Lisa Schiffelers of @FISchmidtLab . Antagonistic camelid #nanobodies are small enoug….

In parellel work, @anja_kopp @Geyer_lab solved the structure of two of our #nanobodies bound to #GSDMD. The Feng Shao lab also used #AAV to express one of our #GSDMD #nanobodies to prevent loss of the #BBB during sepsis.

Special thanks to all the lab members who helped during the revision, including @YM_Tesfamariam, Lea, Stefan, Sabine and Steffen. This work was done at @IIIBonn @UniBonn @UniklinikBonn with funding by @ImmunoSens @sfb_1403 @237Crc .

We think the antagonistic GSDMD #nanobodies enter cells when the first #GSDMD pores form. They prevent any further pore formation and thus -in combination with cellular repair mechanism- prevent cell death by pyroptosis. We are looking forward to test this concept in vivo . .

As #nanobodies enter pyroptotic cells (left), Lisa also tested if recombinant #nanobodies prevent pyroptosis when added to the medium during #inflammasome activation (right). They do!

We learn: 1. The plasma membrane is the first #GSDMD target membrane, 2. Cleavage of #GSDMD is sufficient to allow membrane insertion (no prepore required), and 3. #GSDMD pores are rapidly removed as soon as pores form.

As cells no longer die by #pyroptosis, stabilized, endogenous #GSDMD in the plasma membrane can even stained with antibodies for cleaved GSDMD.

Surprisingly, expression of antagonistic #GSDMD #nanobodies stabilized monomeric GSDMD NTD-mNeonGreen in the plasma membrane.

We had previously tried to use a cool tool from @inflammgenetics to visualize #GSDMD pores in living cells (mNeonGreen remains with the #GSDMD NT), but mostly found GSDMD in intracellular structures .

We raised #alpaca #nanobodies against #GSDMD and found that two cytosolically expressed nanobodies completely shut down pore formation, #pyroptosis, and cytokine release after #inflammasome activation .

RT @TheDohertyInst: ✨Today officially marks 10 years of @TheDohertyInst. From pioneering research on the immune system to our response to….

RT @Synth_Immune: Honored to win the Alessandro Moretta Memorial Award. Fantastic science in a wonderful setting in Greece. @ragoninsti….