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Intracellular absorption underlies collective bacterial tolerance towards an antimicrobial peptide

The collective tolerance towards antimicrobial peptides (APs) is thought to occur primarily through mechanisms associated with live bacterial cells. In contrast to the focus on live cells, we discover that the LL37 antimicrobial peptide kills Escherichia coli, forming a subpopulation of dead cells that absorbs the remaining LL37 into its intracellular space. Combining mathematical modeling with population and single-cell experiments, we show that bacteria absorb LL37 at a timing that coincides with the permeabilization of their cytoplasmic membranes. Furthermore, we show that one bacterial strain can absorb LL37 and protect another strain from killing by LL37. Finally, we demonstrate that the intracellular absorption of LL37 can be reduced using a peptide adjuvant. In contrast to the existing collective tolerance mechanisms, we show that the dead-bacterial absorption of APs is a dynamic process that leads to emergent population behavior, and the work suggests new directions to enhance the efficacy of APs.