Cell surface-associated proteinases in NK cell-mediated cytotoxicity: enhancement of enzyme expression is unique to activation with interferon-alpha.
The human NK cell-mediated cytotoxicity reaction is sensitive to proteinase inhibitors with specificity for chymotrypsin-like enzymes inhibitable by 1-tosylamide 2-phenylethyl chloromethyl ketone (TPCK). Evidence is presented in support of previous data suggesting that this type of cytotoxicity is attributable to enzymes associated with the surface membrane of the NK cell. Activation of the cells with IFN-alpha results in increased cytolytic activity, the suppression of which requires an almost two- to threefold increase in the concentration of proteinase inhibitors. Treatment of NK cells with IFN-alpha results in increased surface binding of [3H]diisopropyl fluorophosphate ([ 3H]DFP). This effect is not inhibited by cycloheximide (50 micrograms/ml), suggesting translocation of preexisting enzymes to the surface membrane. TPCK can compete with [3H]-DFP for binding to the cell surface and can abrogate the increase in [3H]DFP binding observed after IFN-alpha stimulation of the cells. Treatment with IFN-gamma does not increase cell surface-associated proteolytic activity and stimulation with IL-2 results in much smaller increments. The sensitivity of cytotoxicity to proteinase inhibitors is confined to the initial 2-5 min of the reaction. This suggests that cell surface-associated proteinases play a role in the programming of NK cells for lysis, whereas subsequent events may be dependent on secreted enzyme moieties.
Lavie
G
G
Department of Medicine, New York University Medical Center, New York 10016.
Zucker-Franklin
D
D
D007611:Aprotinin / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D003602:Cytotoxicity, Immunologic; D006801:Humans; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000502:physiology*; D008213:Lymphocyte Activation / Q000187:drug effects; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D013997:Time Factors
D007370:Interferon Type I; D007376:Interleukin-2; D011480:Protease Inhibitors; D007371:Interferon-gamma; D007611:Aprotinin; D010447:Peptide Hydrolases