Specific release of proteoglycans from human natural killer cells during target lysis

Schmidt, R. E.; MacDermott, Richard P.; Bartley, G; Bertovich, Michael J.; Amato, David A.; Austen, K. Frank; Schlossman, S F; Stevens, Richard L; Ritz, Jérôme

doi:10.1038/318289a0

Cited by 136 publications

(40 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Exocytosis of the 35S-labeled proteoglycan from the CNK8 cell was also stimulus specific, being 53% upon incubation with the K562 sensitive target, and 3% for the CEM-resistant target. The correlation of proteoglycan release with target cell susceptibility, coupled with the inhibition of proteoglycan release by monocional antibodies that block JTBI8 and CNK8 cytotoxicity indicates that specific NK effector cell receptors and target cell recognition structures are involved in triggering proteoglycan exocytosis (42). The ability to define one of the granule constituents, proteoglycan, exocytosed from the NK cell via receptor-mediated activation should enable the identification of other granule-associated molecules and their possible association with this proteoglycan molecule.…”

Section: Discussionmentioning

confidence: 99%

Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis.

MacDermott¹,

Schmidt²,

Caulfield³

et al. 1985

The Journal of Experimental Medicine

143

View full text Add to dashboard Cite

A clone of natural killer (NK) cells (JTB18) was found to be ultrastructurally similar to peripheral blood large granular lymphocytes (LGL). These cells incorporated [35S]sulfate into cell-associated proteoglycan molecules, which were then isolated by CsCl density gradient centrifugation. As assessed by gel filtration chromatography, the native 35S-labeled proteoglycan and its beta-eliminated 35S-labeled glycosaminoglycans were of Mr approximately 200,000 and 50,000, respectively. The 35S-labeled proteoglycans were resistant to proteolysis, since their Mr were apparently not altered by incubation with either pronase or S. aureus V8 protease. The purified NK cell 35S-labeled proteoglycans were degraded by approximately 90% to 35S-labeled disaccharides with either chondroitinase ABC or AC. High performance liquid chromatographic analysis of the digests revealed these disaccharides to be composed entirely of chondroitin sulfate A (glucuronic acid----N-acetylgalactosamine-4SO4). Whole 35S-labeled cells incubated with chondroitinase ABC failed to release 35S-labeled disaccharides into the supernatant, and x-ray energy-dispersive analysis revealed that sulfur-containing molecules were present in the intracellular granules, thereby localizing the NK cell-associated proteoglycan primarily in the granules of the cell, rather than on the plasma membrane. The 35S-labeled cloned NK cells incubated for 30 min to 4 h with K562 tumor cell targets at a 0.5:1 ratio exocytosed a mean of 49% of the granular 35S-labeled proteoglycans during the first 60 min of the culture. Proteoglycan release was maximal with an effector/target cell ratio of 0.5:1 for JTB18:K562. Significant proteoglycan release from JTB18 NK cells was also obtained with other sensitive target cells such as REX, Molt4, and CEM, but not with cells such as KG1 and Laz156, which have been shown previously to be resistant to killing by this NK cell. Thus, protease-resistant intracellular proteoglycans with chondroitin sulfate A side chains are specifically exocytosed from the granules of human NK effector cells upon contact with sensitive targets, suggesting that these proteoglycans may be involved in the mechanism of cytotoxicity.

show abstract

Section: Discussionmentioning

confidence: 99%

Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis.

MacDermott¹,

Schmidt²,

Caulfield³

et al. 1985

The Journal of Experimental Medicine

143

View full text Add to dashboard Cite

show abstract

“…It has been known for a considerable time that PGs are components of the secretory granule of cytotoxic cells. In 1985 it was reported that NK cells contained a protease-resistant chondroitin sulfate PG species that was released upon incubation of the NK cells with target cells, and the possibility that these PGs are involved in the packaging of cytolytic granule compounds was discussed (16,17). Indeed, a subsequent report suggested that both perforin and granzyme A interacted with PGs at acidic pH, although the interaction with perforin with PGs was weakened at neutral pH (18).…”

mentioning

confidence: 99%

Serglycin-deficient Cytotoxic T Lymphocytes Display Defective Secretory Granule Maturation and Granzyme B Storage

Grujić

Braga

Lukinius

et al. 2005

Journal of Biological Chemistry

109

107

View full text Add to dashboard Cite

Cytotoxic T lymphocytes eliminate infected and tumor cells mainly by perforin/granzyme-induced apoptosis. Earlier studies suggested that serglycin-proteoglycans form macromolecular complexes with granzymes and perforin in the cytotoxic granule. Serglycin-proteoglycans may also be involved in the delivery of the cytolytic machinery into target cells. We have developed a serglycin-deficient mouse strain, and here we studied the importance of serglycin-proteoglycans for various aspects of cytotoxic T lymphocyte function.35 SO 4 2؊ radiolabeling of serglycin-deficient cells demonstrated a dramatic reduction of incorporated label as compared with wild type cells, indicating that serglycin is by far the dominating proteoglycan species produced by the cytotoxic T lymphocyte. Moreover, lack of serglycin resulted in impaired ability of cytotoxic T lymphocytes to produce secretory granule of high electron density, although granule of lower electron density were produced both in wild type and serglycin-deficient cells. The serglycin deficiency did not affect the mRNA expression for granzyme A, granzyme B, or perforin. However, the storage of granzyme B, but not granzyme A, Fas ligand, or perforin, was severely defective in serglycin-deficient cells. Serglycin-deficient cells did not display defects in late cytotoxicity toward target cell lines. Taken together, these results point to a key role for serglycin in the storage of granzyme B and for secretory granule maturation but argue against a major role for serglycin in the apoptosis mediated by cytotoxic T lymphocytes.

show abstract

“…Recently, proteoglycans of the chondroitin sulfate A type have been identified in the granules of NK cells and shown to be released from cells during cytolysis (25,26). As shown here, proteoglycans may also be used as a granule marker for CTLL .…”

Section: Discussionmentioning

confidence: 81%

“…Proteoglycans of CTLL-R8 cell granules can also be visualized on polyacrylamide gels stained with Alcian Blue, which reveal several bands of different molecular masses (profile not shown here). Because proteoglycans have previously been speculated to exert a protective function during cell-mediated killing, tentatively through their binding to PFP/perforin released from cells (25,26), we tested the effects of heparin and chondroitin sulfate on the hemolytic activity mediated by PFP. Only heparin inhibited hemolysis at high concentrations (0.5 mg/ml), whereas chondroitin sulfate (and the three subtypes A, B, and C) produced no effect in the wide concentration range tested in our experiments.…”

Section: Discussionmentioning

confidence: 99%

Dissociation of membrane binding and lytic activities of the lymphocyte pore-forming protein (perforin).

Young¹,

Damiano²,

Dinome³

et al. 1987

The Journal of Experimental Medicine

View full text Add to dashboard Cite

Granules isolated from CTL and NK cells contain a cytolytic pore-forming protein (PFP/perforin). At low temperatures (on ice), PFP binds to erythrocyte membranes without producing hemolysis. Hemolysis occurs when the PFP-bound erythrocytes are warmed up to 37 degrees C, which defines a temperature-dependent, lytic (pore-formation) step distinct from the membrane-binding event. Ca2+ and neutral pH are required for both membrane binding and pore formation by PFP. Serum, LDL, HDL, and heparin inhibit the hemolytic activity of PFP by blocking its binding to lipid membranes. Lysis by PFP that has bound to erythrocyte membranes is no longer susceptible to the effect of these inhibitors. The hemolytic activities associated with intact granules and solubilized PFP show different requirements for Ca2+ and pH, indicating that cytolysis produced by isolated granules may involve an additional step, possibly fusion of granules with membranes. It is suggested that three distinct Ca2+- and pH-dependent events may be involved during cell killing by CTL and NK cells: fusion of cytoplasmic granules of effector cells with their plasma membrane, releasing PFP from cells; binding of the released PFP to target membranes; and insertion of monomers and the subsequent formation of lytic pores in the target membrane. The serum-mediated inhibition of membrane binding by PFP could prevent the accidental injury of bystander cells by cell-released PFP, but would allow cytolysis to proceed to completion once PFP has bound to the target membrane.

show abstract

Specific release of proteoglycans from human natural killer cells during target lysis

Cited by 136 publications

References 17 publications

Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis.

Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis.

Serglycin-deficient Cytotoxic T Lymphocytes Display Defective Secretory Granule Maturation and Granzyme B Storage

Dissociation of membrane binding and lytic activities of the lymphocyte pore-forming protein (perforin).

Contact Info

Product

Resources

About