Degranulation enhances presynaptic membrane packing, which protects NK cells from perforin-mediated autolysis.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101183755 Publication Model: eCollection Cited Medium: Internet ISSN: 1545-7885 (Electronic) Linking ISSN: 15449173 NLM ISO Abbreviation: PLoS Biol Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science, [2003]-

Cell Degranulation*; Killer Cells, Natural/*physiology ; Membrane Lipids/*metabolism ; Perforin/*metabolism; Cell Line, Tumor ; Cell Survival ; Healthy Volunteers ; Humans ; Ketocholesterols ; Primary Cell Culture

Competing Interests: The authors have declared that no competing interests exist.
Natural killer (NK) cells kill a target cell by secreting perforin into the lytic immunological synapse, a specialized interface formed between the NK cell and its target. Perforin creates pores in target cell membranes allowing delivery of proapoptotic enzymes. Despite the fact that secreted perforin is in close range to both the NK and target cell membranes, the NK cell typically survives while the target cell does not. How NK cells preferentially avoid death during the secretion of perforin via the degranulation of their perforin-containing organelles (lytic granules) is perplexing. Here, we demonstrate that NK cells are protected from perforin-mediated autolysis by densely packed and highly ordered presynaptic lipid membranes, which increase packing upon synapse formation. When treated with 7-ketocholesterol, lipid packing is reduced in NK cells making them susceptible to perforin-mediated lysis after degranulation. Using high-resolution imaging and lipidomics, we identified lytic granules themselves as having endogenously densely packed lipid membranes. During degranulation, lytic granule-cell membrane fusion thereby further augments presynaptic membrane packing, enhancing membrane protection at the specific sites where NK cells would face maximum concentrations of secreted perforin. Additionally, we found that an aggressive breast cancer cell line is perforin resistant and evades NK cell-mediated killing owing to a densely packed postsynaptic membrane. By disrupting membrane packing, these cells were switched to an NK-susceptible state, which could suggest strategies for improving cytotoxic cell-based cancer therapies. Thus, lipid membranes serve an unexpected role in NK cell functionality protecting them from autolysis, while degranulation allows for the inherent lytic granule membrane properties to create local ordered lipid "shields" against self-destruction.

Comment in: PLoS Biol. 2021 Aug 4;19(8):e3001339. doi: 10.1371/journal.pbio.3001339. (PMID: 34347789)

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R01 AI067946 United States AI NIAID NIH HHS; R37 AI067946 United States AI NIAID NIH HHS

Dryad 10.5061/dryad.hqbzkh1f4

0 (Ketocholesterols)
0 (Membrane Lipids)
126465-35-8 (Perforin)
O7676FE78M (7-ketocholesterol)

Date Created: 20210803 Date Completed: 20211118 Latest Revision: 20240811

20250114

PMC8330931

10.1371/journal.pbio.3001328

34343168