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CD8α and CD70 mark human natural killer cell populations which differ in cytotoxicity

ABSTRACT: Natural Killer (NK) cells are innate immune cells that can directly detect and kill cancer cells. Understanding the molecular determinants regulating human NK cell cytotoxicity could help harness these cells for cancer therapies. To this end, we compared the transcriptome of NK cell clones derived from human peripheral blood, which were strongly or weakly cytotoxic against 721.221 and other target cells. After one month of culture, potent NK cell clones showed a significant upregulation in genes involved in cell cycle progression, suggesting that proliferating NK cells were particularly cytotoxic. Beyond two months of culture, NK cell clones which were strongly cytotoxic varied in their expression of 28 genes, including CD8Α and CD70; NK cells with high levels of CD70 expression were weakly cytotoxic while high CD8Α correlated with strong cytotoxicity. Thus, NK cells were cultured and sorted for expression of CD70 and CD8α, and in accordance with the transcriptomic data, CD70 + NK cells showed low cytotoxicity against 721.221 and K562 target cells. Cytotoxicity of CD70 + NK cells could be enhanced using blocking antibodies against CD70, indicating a direct role for CD70 in mediating low cytotoxicity. Furthermore, time-lapse microscopy of NK cell-target cell interactions revealed that CD8α + NK cells have an increased propensity to sequentially engage and kill multiple target cells. Thus, these two markers relate to NK cell populations which are capable of potent killing (CD70 -) or serial killing (CD8α + ).

ORGANISM(S): Homo sapiens

PROVIDER: GSE289212 | GEO | 2025/02/11

REPOSITORIES: GEO

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Project description:The paper describes a model of ADCC. Created by COPASI 4.26 (Build 213) This model is described in the article: A mathematical model of antibody-dependent cellular cytotoxicity (ADCC) F. Hoffman, D. Gavaghan, J. Osborne, I.P. Barrett, T. You, H. Ghadially, R. Sainson, R.W. Wilkinson, H.M. Byrne Journal of Theoretical Biology 436 (2018) 39–50 Abstract: Immunotherapies exploit the immune system to target and kill cancer cells, while sparing healthy tis- sue. Antibody therapies, an important class of immunotherapies, involve the binding to specific antigens on the surface of the tumour cells of antibodies that activate natural killer (NK) cells to kill the tu- mour cells. Preclinical assessment of molecules that may cause antibody-dependent cellular cytotoxicity (ADCC) involves co-culturing cancer cells, NK cells and antibody in vitro for several hours and measuring subsequent levels of tumour cell lysis. Here we develop a mathematical model of such an in vitro ADCC assay, formulated as a system of time-dependent ordinary differential equations and in which NK cells kill cancer cells at a rate which depends on the amount of antibody bound to each cancer cell. Numerical simulations generated using experimentally-based parameter estimates reveal that the system evolves on two timescales: a fast timescale on which antibodies bind to receptors on the surface of the tumour cells, and NK cells form complexes with the cancer cells, and a longer time-scale on which the NK cells kill the cancer cells. We construct approximate model solutions on each timescale, and show that they are in good agreement with numerical simulations of the full system. Our results show how the processes involved in ADCC change as the initial concentration of antibody and NK-cancer cell ratio are varied. We use these results to explain what information about the tumour cell kill rate can be extracted from the cytotoxicity assays. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models . To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Project description:Natural Killer (NK) cells are phenotypically and functionally diverse lymphocytes that recognize and kill cancer cells. The susceptibility of target cancer cells to NK cell-mediated cytotoxicity depends on the strength and balance of regulatory (activating/inhibitory) ligands expressed on target cell surface. We performed gene expression arrays to determine patterns of NK cell ligands associated with B-cell non-Hodgkin lymphoma (b-NHL). Microarray analyses revealed differential upregulation of a multitude of NK-activating and costimulatory ligands across varied lymphoma cell lines and b-NHL cells including ULBP1, CD72, CD48 and SLAMF6. To correlate genetic signatures with functional anti-lymphoma activity, we developed a dynamic and quantitative cytotoxicity assay in an integrated microfluidic droplet array. This assay facilitated the examination of transient interactions between NK cells and target lymphoma cells and also the functional heterogeneity in NK cell effector outcomes at single-cell level. Individual NK cells and target lymphoma cells were co-encapsulated in picoliter-volume droplets and imaged over time to quantify heterotypic cell-cell interaction. We identified significant variability in NK-lymphoma cell contact duration, frequency and subsequent cytolysis. The death of lymphoma cells undergoing single contact with NK cells occurred faster than cells that made multiple short contacts. NK cells also killed target cells in droplets via contact-independent mechanisms that partially relied on calcium-dependent processes and perforin secretion, but not on cytokines (IFN-γ or TNF-α). We extended this technique to characterize cytolysis of primary cells from b-NHL patients. Diffuse large B-cell lymphoma cells from two patients showed similar contact durations with NK cells; primary Burkitt lymphoma cells made longer contacts and were lysed at later times. We further tested the cytotoxic efficacy of NK-92, a continuously growing NK cell line being investigated as an anti-tumor therapy, using our droplet-based bioassay. NK-92 cells were found to be more efficient in killing b-NHL cells compared with primary/fresh NK cells, requiring shorter contacts for faster killing activity. Taken together, our combined genetic and microfluidic analysis demonstrate significant heterogeneity in the dynamic interaction and associated cytotoxicity between NK cellular therapy and b-NHL cells.

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CD8α and CD70 mark human natural killer cell populations which differ in cytotoxicity

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