Generation of monoclonal antibodies to a human natural killer clone. Characterization of two natural killer-associated antigens, NKH1A and NKH2, expressed on subsets of large granular lymphocytes.

Hercend, Thierry; Griffin, JD; Bensussan, Armand; Schmidt, Reinhold; Edson, Mary Ann; Brennan, Agnes; Murray, Christine; Daley, John; Schlossman, S F; Ritz, Jérôme

doi:10.1172/jci111794

Cited by 254 publications

(95 citation statements)

References 30 publications

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“…The activated PBMC were then washed four times to remove the mitogens, and were resuspended in RPMI 1640 supplemented with 2.5% human AB serum. After 40 h of incubation at 37°C, the cells were pelleted at 500 g, and the LCM was sterilized by passage through 0.45 #m filters and stored at -70°C until it was used for cloning or subcloning the JTB18 cells (21)(22)(23)(24). All NK cells used in these studies were subcloned at least four times at 100 cells/well on a feeder layer of allogeneic irradiated PBMC plus irradiated EBV-transformed B cells.…”

Section: Methodsmentioning

confidence: 99%

“…Cytotoxicity assays with human tumor lines used as targets were performed according to a standard chromium-release method (21)(22)(23)(24). The K562 cell line was established from a patient with chronic myeiogenous leukemia and the KG1 cell line from a patient with acute myeloblastic leukemia.…”

Section: Radiolabeling Of Nk-cloned Cells and Characterization Of Celmentioning

confidence: 99%

“…The negative charge characteristics of these intracellular proteoglycans make them particularly well suited as storage and carrier molecules for basically charged proteases and other proteins that are present in secretory granules and are exocytosed in response to a specific stimulus. We investigated the human NK clone JTB18, which has the plasma membrane phenotype of human peripheral blood NK cells (21), and a defined target cell specificity (22), to determine whether it possesses proteoglycans in its granules. We determined that this cloned human NK cell contains a cell-associated chondroitin sulfate A proteoglycan which is localized to the granules of the cell and is specifically exocytosed when the NK cell lyses susceptible tumor cells.…”

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confidence: 99%

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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

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143

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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.

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Section: Methodsmentioning

confidence: 99%

Section: Radiolabeling Of Nk-cloned Cells and Characterization Of Celmentioning

confidence: 99%

mentioning

confidence: 99%

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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

Self Cite

143

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“…In humans, NK lymphocytes are phenotypically characterized by the expression of CD56, an isoform of the neural cell adhesion molecule, and by the absence of CD3 (1,4,5). In peripheral blood (PB), 3 two subsets of NK cells were identified, namely, a CD56 dim and a CD56 bright population corresponding to ϳ95 and 5% of the total circulating NK pool, respectively.…”

Section: N Atural Killer Cells Are a Class Of Lymphocytes Involved Inmentioning

confidence: 99%

A Soluble Form of the MHC Class I-Specific CD160 Receptor Is Released from Human Activated NK Lymphocytes and Inhibits Cell-Mediated Cytotoxicity

Giustiniani

Marie‐Cardine

Bensussan

2007

The Journal of Immunology

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CD160 is a GPI-anchored lymphocyte surface receptor in which expression is mostly restricted to the highly cytotoxic CD56dimCD16+ peripheral blood NK subset. We previously reported that MHC class I (MHC-I) molecules bind to CD160 receptors on circulating NK lymphocytes and that this interaction triggers their cytotoxic activity and cytokine production. We also observed that CD160 surface expression on NK cells is down-modulated upon activation with PMA or IL-2. In this study, we further report that short-time incubation of NK lymphocytes with IL-15 converts the membrane-bound CD160 to a soluble form through a proteolytic cleavage involving a metalloprotease. Thus, CD160 is no longer detected at the cell surface, but can be immunoprecipitated from the NK cell culture medium. Interestingly, CD160 transcript remains highly expressed during the process of protein shedding. In addition, we demonstrate that CD160 mRNA synthesis can be induced in CD56bright separated lymphocytes following exposure to IL-15. By producing a Flag-tagged soluble CD160 protein, we establish that its binding to MHC-I molecules results in the inhibition of the cytotoxic CD8+ T lymphocyte activity and of the CD160-mediated NK cell cytotoxicity. Thus, we show that activated NK lymphocytes release a soluble form of CD160 that functionally impairs the MHC-I-specific cytotoxic CD8+ T lymphocyte responsiveness.

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“…Despite their homogeneous morphology, mature NK effector cells are a phenotypically heterogeneous subset of cells that bear a large number of surface antigens (22,27,37). It has been clearly established that the majority (90% ) of peripheral blood lymphocytes mediating MHC-unrestricted cytotoxicity express the CD16 (Fc gamma-111) (18,24,27,31) and CD56 (neural cell adhesion molecule) antigens (12,17). CD16 positive lymphocytes may also coexpress (20-50% ) the CD57 surface antigen, although 0 1995 Wiley-Liss, Inc. a percentage of CD57 negative cells can mediate NK activity (22,27,28).…”

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Evidence of a selective depletion of a CD16⁺ CD56⁺ CD8⁺ natural killer cell subset during HIV infection

et al. 1995

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Three-color flow cytometric analysis of CD16+ natural killer (NK) cells was assessed in HIV seropositive patients and healthy heterosexual controls. A selective depletion of lymphocytes with the CD16+ NK phenotype was found among the HIV+ infected patients. When the CD16 lymphocyte subset was further evaluated by three-color flow cytometry, cells bearing both the CD8 and CD56 antigens were significantly decreased. Analysis of activation antigens revealed a large proportion of CD16+ NK cells from HIV+ patients expressed HLA-DR, but this did not correlate with CD25 (IL-2 receptor) expression. The overall loss of the CD8 and CD56 antigens among the NK population with an increase in activation status may be due to differential loss of the NK cell subsets or, alternatively, to the loss of immunoregulatory cytokines, which have been shown to be important in maintaining NK activity. Whether these changes in the NK compartment may influence the outcome of individuals with HIV disease still remains an open question but is an important issue when performing phenotypic analysis of HIV+ subjects. o 1995 Wiley-Liss, Inc.

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Generation of monoclonal antibodies to a human natural killer clone. Characterization of two natural killer-associated antigens, NKH1A and NKH2, expressed on subsets of large granular lymphocytes.

Cited by 254 publications

References 30 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.

A Soluble Form of the MHC Class I-Specific CD160 Receptor Is Released from Human Activated NK Lymphocytes and Inhibits Cell-Mediated Cytotoxicity

Evidence of a selective depletion of a CD16⁺ CD56⁺ CD8⁺ natural killer cell subset during HIV infection

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Generation of monoclonal antibodies to a human natural killer clone. Characterization of two natural killer-associated antigens, NKH1A and NKH2, expressed on subsets of large granular lymphocytes.

Cited by 254 publications

References 30 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.

A Soluble Form of the MHC Class I-Specific CD160 Receptor Is Released from Human Activated NK Lymphocytes and Inhibits Cell-Mediated Cytotoxicity

Evidence of a selective depletion of a CD16+ CD56+ CD8+ natural killer cell subset during HIV infection

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Evidence of a selective depletion of a CD16⁺ CD56⁺ CD8⁺ natural killer cell subset during HIV infection