Isolation of human NK cells by density gradient centrifugation

doi:10.1016/0022-1759(80)90133-7

Journal of Immunological Methods

Volume 36, Issues 3–4, 16 September 1980, Pages 285-291

https://doi.org/10.1016/0022-1759(80)90133-7 Get rights and content

Abstract

Sedimentation characteristics of human NK cells in discontinuous Percoll density gradients were studied. NK activity against the leukaemic cell line K-562 peaked in a single low density fraction mainly consisting of large granular lymphocytes previously shown to be the principal NK cells in human peripheral blood. Percoll density gradient centrifugation provides a useful tool for analysis of human NK cells and their relationship to other blood mononuclear cells.

References (13)

R.B. Herberman et al.
Adv. Cancer Res.
(1978)
T. Timonen et al.
Cell. Immunol.
(1979)
T. Timonen et al.
Cell. Immunol.
(1979)
L.C. Andersson et al.
Int. J. Cancer
(1979)
A. Böyum
Scand. J. Clin. Lab. Invest.
(1968)
J.E. De Vries et al.
Int. J. Cancer
(1980)

There are more references available in the full text version of this article.

Cited by (362)

Bracing NK cell based therapy to relegate pulmonary inflammation in COVID-19
2021, Heliyon
Citation Excerpt :
The potential source to isolate NK cells includes peripheral blood and secondary lymphoid organs (lymph node, tonsil, spleen, and lymph) respectively. Notably, the peripheral blood results in more quantities of NK cells in comparison to other lymphoid organs [57]. The method of NK cell isolation from peripheral blood is described here.
The contagiosity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has startled mankind and has brought our lives to a standstill. The treatment focused mainly on repurposed immunomodulatory and antiviral agents along with the availability of a few vaccines for prophylaxis to vanquish COVID-19. This seemingly mandates a deeper understanding of the disease pathogenesis. This necessitates a plausible extrapolation of cell-based therapy to COVID-19 and is regarded equivalently significant. Recently, correlative pieces of clinical evidence reported a robust decline in lymphocyte count in severe COVID-19 patients that suggest dysregulated immune responses as a key element contributing to the pathophysiological alterations. The large granular lymphocytes also known as natural killer (NK) cells play a heterogeneous role in biological functioning wherein their frontline action defends the body against a wide array of infections and tumors. They prominently play a critical role in viral clearance and executing immuno-modulatory activities. Accumulated clinical evidence demonstrate a decrease in the number of NK cells in circulation with or without phenotypical exhaustion. These plausibly contribute to the progression of pulmonary inflammation in COVID-19 pneumonia and result in acute lung injury. In this review, we have outlined the present understanding of the immunological response of NK cells in COVID-19 infection. We have also discussed the possible use of these powerful biological cells as a therapeutic agent in view of preventing immunological harms of SARS-CoV-2 and the current challenges in advocating NK cell therapy for the same.
Historical overview on the morphological characterization of large granular lymphocytes/natural killer cells
2017, Immunology Letters
Citation Excerpt :
LGLs are larger than the typical lymphocytes (10–12 μm) with a large amount of cytoplasm containing peroxidase-negative granules. Most NK cells have LGL morphological aspects [19]. However, not all NK cells have LGL morphology and not all LGL cells are NK cells.
Natural Killer (NK) cells were discovered when research on T cells produced an unexplained background, or “natural” cytotoxicity in non-immunized mice. The previously unknown cells were thus named “natural” killer cells, or NK cells. They are roughly defined by a lack of the T cell marker CD3 and a presence of CD56 (or NCAM, neural cell adhesion molecule) and correspond to the large granular lymphocytes (LGLs). This article is focused to the description the morphological features of these cells and of the LGLs and to the fundamental contribution of the Italian scientist Carlo E. Grossi to the ultrastructural characterization of these cells.
Expansion of NK cells by engineered K562 cells co-expressing 4-1BBL and mMICA, combined with soluble IL-21
2014, Cellular Immunology
Citation Excerpt :
Human natural killer (NK) cells comprise ≈15% of all circulating lymphocytes [1].
NK cells hold promise for protecting hosts from cancer and pathogen infection through direct killing and expressing immune-regulatory cytokines. In our study, a genetically modified K562 cell line with surface expression of 4-1BBL and MICA was constructed to expand functional NK cells in vitro for further adoptive immunotherapy against cancer. After a long-term up to 21 day co-culture with newly isolated peripheral blood mononuclear cells (PBMCs) in the presence of soluble IL-21 (sIL-21), notable increase in proportion of expanded NK cells was observed, especially the CD56^brightCD16⁺ subset. Apparent up-regulation of activating receptors CD38, CD69 and NKG2D was detected on expanded NK cells, so did inhibitory receptor CD94; the cytotoxicity of expanded NK cells against target tumor cells exceeded that of NK cells within fresh PBMCs. The intracellular staining showed expanded NK cells produced immune-regulatory IFN-γ. Taken together, we expanded NK cells with significant up-regulation of activating NKG2D and moderate enhancement of cytotoxicity, with IFN-γ producing ability and a more heterogeneous population of NK cells. These findings provide a novel perspective on expanding NK cells in vitro for further biology study and adoptive immunotherapy of NK cells against cancer.
Histamine in cancer immunotherapy: A preclinical background
2002, Seminars in Oncology
Natural killer (NK) cells and T cells are the primary targets of interleukin-2 (IL-2) and other cytokines used in cancer immunotherapy. However, these tumoricidal lymphocytes are frequently dysfunctional or apoptotic when residing within melanomas and other solid cancers. This phenomenon-tumor-induced immunosuppression-is poorly understood and conceivably limits the efficiency of strategies aiming at activating lymphocyte-mediated antitumor immunity. Recent studies imply that reactive oxygen species (oxygen radicals), produced by tumor-infiltrating monocyte/macrophages, may contribute to the state of lymphocyte inhibition in neoplastic tissue. Histamine, acting via H2-type histamine receptors on monocyte/macrophages, suppresses the activity of a key enzyme in oxygen radical formation, the NADPH oxidase. By this mechanism, histamine protects NK cells and T cells against oxygen radical-induced dysfunction and apoptosis, and also maintains their activation by IL-2 and other lymphocyte activators. In this review, these properties of histamine are discussed in relation to the current use of histamine as an adjunct to IL-2 in metastatic melanoma and other malignant diseases. Semin Oncol 29 (suppl 7):35-40. Copyright 2002, Elsevier Science (USA). All rights reserved.
Aptamer-Based Traceless Multiplexed Cell Isolation Systems
2022, ACS Applied Materials and Interfaces
Natural killer cell-mediated immunopathology in recurrent pregnancy losses
2022, Exploration of Immunology

View all citing articles on Scopus

View full text

Isolation of human NK cells by density gradient centrifugation

Abstract

Adv. Cancer Res.

Cell. Immunol.

Cell. Immunol.

Int. J. Cancer

Scand. J. Clin. Lab. Invest.

Int. J. Cancer