Engagement of CD160 receptor by HLA-C is a triggering mechanism used by circulating natural killer (NK) cells to mediate cytotoxicity
Philippe Le BouteillerAlíz BarakonyiJérôme GiustinianiFrançoise LenfantAnne Marie‐CardineMaryse Aguerre‐GirrMagali RabotI HilgertFathia Mami‐ChouaibJulie TabiascoLaurence BoumsellArmand Bensussan
134
Citation
30
Reference
10
Related Paper
Citation Trend
Abstract:
Circulating human natural killer (NK) lymphocytes have been functionally defined by their ability to exert cytotoxic activity against MHC class I-negative target cell lines, including K562. Therefore, it was proposed that NK cells recognized the “missing self.” We show here that the Ig-like CD160 receptor expressed by circulating CD56 dim+ NK cells or IL-2-deprived NK cell lines is mainly involved in their cytotoxic activity against K562 target cells. Further, we report that HLA-C molecules that are constitutively expressed by K562 trigger NK cell lysis through CD160 receptor engagement. In addition, we demonstrate, with recombinant soluble HLA-Cw3 and CD160 proteins, direct interaction of these molecules. We also find that CD158b inhibitory receptors partially interfere with CD160-mediated cytotoxicity, whereas CD94/CD159a and CD85j have no effect on engagement with their respective ligands. Thus, CD160/HLA-C interaction constitutes a unique pathway to trigger NK cell cytotoxic activity.Keywords:
K562 cells
Lymphokine-activated killer cell
The natural killer cell cytotoxicity was studied in the 4 hour and 18 hour cytotoxic reaction with cultured human tumour cells (K562 and lung adenocarcinoma) labelled by 51Cr. The natural killer cell activity of cancer patients was lowered only in 4 hour cytotoxic reaction with target cells K562. The natural and recombinant leucocyte interferon and natural immune interferon stimulated in vitro the activity of natural killer cells in most of the experiments (80.5 %) in the 4 hour and 18 hour cytotoxic reactions both from patients with initially lowered level, and with normal or high level of cytotoxic action of natural killer cells. In some experiments interferon preparations did not significantly influence the lysis of tumour cells or even suppressed the activity of effector cells. The heterogeneity of the population of natural killer cells and different immunomodulating effects of interferon were discussed.
K562 cells
Lymphokine-activated killer cell
Cite
Citations (0)
Natural killer (NK) cells form part of the vertebrate defence against viruses and tumours, but show only limited specificity. The molecule(s) recognized by NK cells on target cells are at present unknown. Major histocompatibility complex (MHC) class I antigen concentration on target cells is inversely correlated with NK cell lysis. Here we show that MHC class I‐unassociated β 2 ‐microglobulin (β 2 ‐m) expression is involved in NK cell‐target cell interaction. Two human MHC class I negative cell lines. Daudi and K562, are differentially susceptible to NK cell lysis. Daudi cells are β 2 ‐m‐negative and resistant to NK lysis, K562 are β 2 ‐m‐positive and highly susceptible to lysis by NK cells. Interferon (IFN) treatment augments β 2 ‐m expression and NK lysis of K562, but not in Daudi cells. NK cell lysis of K562, but not YAC‐1 cells, can be inhibited by monoclonal anti‐human β 2 ‐m antibody. Furthermore, susceptibility of mouse embryo fibroblasts (MEF) to NK lysis can be increased by infection with recombinant vaccinia virus expressing the human β 2 ‐m gene.
K562 cells
Beta-2 microglobulin
Lymphokine-activated killer cell
NK-92
BETA (programming language)
Cite
Citations (22)
Lymphocytes isolated from human tonsils are capable of lysing cells of the natural killer (NK)-susceptible line K562. Although weak compared with that of autochthonous peripheral lymphocytes, cytotoxicity is invariably manifest at high effector-to-target ratios. Like peripheral NK cells, tonsillar cytotoxic lymphocytes possess a low buoyant density, which makes possible their partial enrichment from non-cytotoxic cells by centrifugation on discontinuous Percoll gradients. However, examination of cytotoxic fractions indicates that, unlike blood lymphocyte-mediated cytotoxicity, effector function is not associated with the presence of large granular lymphocytes. Furthermore, a functional distinction from classical NK cells is apparent since, although tonsillar cytotoxicity is significantly enhanced after exposure to supernatants from polyclonally activated allogeneic tonsils, pretreatment with lymphoblastoid (Namalva) interferon (IFN-alpha) at doses shown to potentiate maximally the cytotoxicity of peripheral blood lymphocytes fails to influence reactivity. These data provide preliminary evidence for the existence of, at least limited, heterogeneity among human NK cells.
Percoll
K562 cells
Lymphokine-activated killer cell
Lymphoblast
Cite
Citations (8)
This report demonstrates that in vitro activation of human cells with the β-galactoside-specific lectin from mistletoe (ML-I) or interleukin-2 (IL-2) results in different patterns of activation and function of cytotoxic cells. It is now well established that natural killer (NK) and lymphokine-activated killer (LAK) cytotoxicity is mainly mediated by resting (NK) and IL-2-activated (LAK) CD56-positive (+) cells respectively. Culture of peripheral blood lymphocytes (PBL) for 3 days with ML-I led to expansion and activation of T cells which demonstrated NK-and LAK-like cytotoxicity. T lymphocyte subset analysis revealed that in total PBL, ML-I preferentially stimulated and expanded CD8+ T cells which mediated the cytotoxic effect. Incubation of highly purified CD8+ T cells alone with ML-I did not lead to induction of cytotoxicity, which required the presence of both CD4+ and CD 14+ (monocytes) cells, suggesting that ML-I does not exert a direct effect on CD8+ T cells. Activation of PBL with both ML-I and IL-2 resulted in simultaneous induction of T and CD56+ cell-mediated NK and LAK cytotoxicity. These data suggest that treatment with ML-I and DL-2 might provide an approach to induce maximum cytotoxicity against tumors and to recruit both T and NK cells for tumor therapy.
Lymphokine-activated killer cell
Cite
Citations (30)
Supernatants from unstimulated CD16+ natural killer (NK) cells or from CD16+ NK cells cocultured with K562 tumor cells (to generate NK cytotoxic factor) were both cytotoxic to target cells. Interleukin 2 stimulation of the CD16+ NK cells in the absence of tumor cell stimulation resulted in supernatants which mediated an increased cytotoxicity as compared to the unstimulated supernatants. The cytotoxic activity was recovered in the chloroform fraction of a Bligh-Dyer lipid extraction suggesting that the toxic moiety in the CD16+ NK cell-derived supernatants might be a lipid. Separation of the cytotoxic supernatants into Mr less than 10,000 and Mr greater than 10,000 fractions revealed that the Mr less than 10,000 fraction of both supernatants had no detectable protein but retained cytotoxicity equal to that of the matched unfractionated supernatant. For convenience, we refer to this lipid-like cytotoxin in the Mr less than 10,000 fraction of the supernatants from unstimulated CD16+ NK cells as lipotoxin (LTX) and the cytotoxin in the Mr less than 10,000 fraction of supernatant from interleukin 2 stimulated CD16+ NK cells as LTX*. Increasing concentrations of LTX and LTX* caused a dose related increase in cytotoxicity. Both LTX and LTX* mediated killing as early as 18 h and their cytotoxicity was not significantly affected by heating at 56 degrees for 2 h or by freezing and thawing. Heating at 63 degrees C resulted in a decrease in cytotoxic activity of 10 to 20%. The less than 10,000 dalton fraction of supernatants from both unstimulated and interleukin 2 stimulated CD3- cells (a crude NK cell population) mediated greater cytotoxicity than the CD3+ cell supernatants, and the majority of cytotoxicity from the CD3- cell supernatants was recovered in this fraction. Thus, NK cells were more efficient producers of the lipid-like cytotoxin than T-cells but whether LTX made by NK cells can also be made by T-cells remains to be determined. We propose that lipotoxin: (a) coexists with protein cytotoxins in NK cell supernatant preparations; (b) mediates significant cytotoxicity when separated from proteinaceous cytotoxins; (c) is responsible for the spontaneously secreted cytotoxic activity observed by others; (d) is distinct from previously reported proteinaceous cytotoxins, e.g., NK cytotoxic factor, tumor necrosis factor alpha, and cytolysin/perforin; (e) accounts for the lipophilic nature of cytotoxic factor activity in NK cell supernatants; and (f) causes the cytotoxic activity observed in a small molecular weight fraction of stimulated NK cell supernatants.
CD16
Lymphokine-activated killer cell
K562 cells
Cite
Citations (2)
HIV has evolved several strategies to evade recognition by the host immune system including down-regulation of major histocompatibility complex (MHC) class I molecules. However, reduced expression of MHC class I molecules may stimulate natural killer (NK) cell lysis in cells of haematopoietic lineage. Here, we describe how HIV counteracts stimulation of NK cells by stabilizing surface expression of the non-classical MHC class I molecule, HLA-E. We demonstrate enhanced expression of HLA-E on lymphocytes from HIV-infected patients and show that in vitro infection of lymphocytes with HIV results in up-regulation of HLA-E expression and reduced susceptibility to NK cell cytotoxicity. Using HLA-E transfected K-562 cells, we identified the well-known HIV T-cell epitope p24 aa14-22a as a ligand for HLA-E that stabilizes surface expression of HLA-E, favouring inhibition of NK cell cytotoxicity. These results propose HIV-mediated up-regulation of HLA-E expression as an additional evasion strategy targeting the antiviral activities of NK cells, which may contribute to the capability of the virus in establishing chronic infection.
Lymphokine-activated killer cell
Cite
Citations (157)
Study on the cytotoxic activity of NK-92 cells against human nasopharyngeal carcinoma cells in vitro
Objective To study cytotoxic activity of NK-92 cells against human nasopharyngeal carcinoma cells and effect of irradiation on NK-92 cytotoxicity in vitro.Methods NK-92 cells were mixed with ~(51)Cr-labeled CNE or K562 cells at different effector-to-target ratios for a 4 h~(51)Cr-release assay.On the other hand,NK-92 cells were irradiated at 0 and 400 cGy and tested after two days for cytotoxicity against K562 and CNE cells.Results NK-92 cells displayed cytotoxicity against CNE with a mean specific lysis of 36% and 42% at 10∶1 and 20∶1 E:T ratios in vitro,while these levels at 1∶1(11 %) and 5∶1(24 %) E:T ratios were lower;The cytotoxicity of NK-92 against K562 ranged from 35 % to 70 %.NK-92 cells maintained up to 8 %～38 % cytotoxicity against CNE and 31 %～62% cytotoxicity against K562 when irradiated with 400 cGy after two days.Conclusion In vitro NK-92 cells have cytotoxic activity against CNE cells at suitable E:T ratios,and substantial cytotoxicity can be retained up to irradiation dose of 400cGy.
K562 cells
Lymphokine-activated killer cell
Cite
Citations (0)
Several cell clones derived from cell lines obtained from a rat thyroid carcinoma, induced by in vivo injection of the Kirsten murine sarcoma virus into the thyroid gland, and from its spontaneous lung metastases were analysed for their major histocompatability complex (MMC) class I antigen expression. The susceptibility lo natural killer (NK) cell lysis of these clones, differing in their levels of MHC class I antigen expression, was determined and found to vary inversely with the target cell MHC level, confirming numerous reports of the literature. We then tried to localize the step of the multistage natural cytotoxic process, in which class I antigens could interfere, and tested first whether lymphokine(IL‐2) activation of the killer (LAK) cells could overcome the differences in MHC class I expression of target cells. As this did not appear lo be the case, we studied the binding step by cither a cold target inhibition assay and a target binding assay and found that target cells expressing class I antigens show a lower competitive capacity for effector cells than targets not expressing such antigens, indicating that this interference may occur, at least in our system, in the binding step of the cytotoxic process.
Cite
Citations (11)
K562 cells
Phytohaemagglutinin
Lymphokine-activated killer cell
Cite
Citations (5)