Studies on the mechanism of the human natural killer cell lethal hit: evidence for transfer of protease-sensitive structures requisite for target cell lysis.
These studies analyze the effects of various enzymes on the terminal, killer cell-independent (KCIL) stages of the human natural killer (NK) cytolytic mechanism. The addition of trypsin (T), chymotrypsin (CT), or papain (P) to standard NK reaction mixtures (PBL or LGL and K562 cells) completely ablated cytolytic activity, whereas collagenase was ineffective. Inhibition by T was reversed by preincubation with soybean trypsin inhibitor (SBTI) or fetal calf serum, indicating that the inhibition was indeed due to T. Kinetic analysis with the Ca++ pulse experiment indicated that T, CT, and P inhibited lysis well beyond the Ca++-dependent (EDTA-sensitive) stages and essentially stopped further 51Cr release at the time of addition. This observation was confirmed by the ability of T, CT, and P to block lysis during KCIL of programmed K562 targets that were detached from NK cells by EDTA and were suspended in dextran-containing media. The lysis of K562 cells by natural killer cell-derived cytotoxic factors (NKCF) was also blocked by T and CT but not by P. Inhibition of NKCF activity by T could be reversed by SBTI or fetal calf serum. The ability of T, CT, or P to inhibit the lysis of "programmed" K562 targets during KCIL indicates that the NK lethal hit is an active process mediated by protease-sensitive structures, possibly NKCF, delivered to the target cell by the NK cell during the Ca++-dependent programming steps.
D001667:Binding, Competitive; D001790:Blood Physiological Phenomena; D002918:Chymotrypsin / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic / Q000187:drug effects*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D008233:Lymphotoxin-alpha / Q000378:metabolism / Q000494:pharmacology*; D010206:Papain / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000494:pharmacology*; D014357:Trypsin / Q000494:pharmacology; D014361:Trypsin Inhibitors / Q000494:pharmacology
D008233:Lymphotoxin-alpha; D014361:Trypsin Inhibitors; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D014357:Trypsin; D010206:Papain