Likewise, a Listeria mutant (ΔactA) that could no longer disseminate to neighboring cells (by ActA-dependent actin tails), even in combination with the aforementioned MAMPs, was unable to trigger NLRP6 activation compared to wild-type Listeria. Therefore, it was not primary HIEC cell infection alone that could trigger NLRP6, but rather spreading of bacteria to neighboring cells, and more specifically their escape from a double membrane vacuole into the cytosol during secondary infection. Here, NLRP6 is linked to infection.