Studies on NK cell purification by negative selection in human peripheral blood

Biotherapy ◽  
1992 ◽  
Vol 5 (1) ◽  
pp. 11-20 ◽  
Author(s):  
Françoise Farace ◽  
Anne-Marie Le Ridant ◽  
Bernard Escudier ◽  
Thierry Hercend ◽  
Frédéric Triebel
Keyword(s):  
Peripheral Blood ◽  
Negative Selection ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Cell Purification

Glycodelin-A stimulates the conversion of human peripheral blood CD16−CD56bright NK cell to a decidual NK cell-like phenotype

2018 ◽  
Vol 34 (4) ◽  
pp. 689-701 ◽  
Author(s):  
Cheuk-Lun Lee ◽  
Madhavi Vijayan ◽  
Xia Wang ◽  
Kevin K W Lam ◽  
Hannu Koistinen ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Glycodelin A

Efficient and Reproducible Retroviral Infection of Primary Human Natural Killer Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1348-1348
Author(s):  
Brian Becknell ◽  
Rossana Trotta ◽  
Jianhua Yu ◽  
Wei Ding ◽  
Hsiaoyin C. Mao ◽  
...  
Keyword(s):  
Nk Cells ◽  
Natural Killer ◽  
Peripheral Blood ◽  
Targeted Delivery ◽  
Magnetic Beads ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Cell Physiology ◽  
Gene Products ◽  
Successful Infection

Abstract Molecular characterization of human natural killer (NK) cells will require targeted gene delivery to inhibit and activate specific signaling pathways, yet to our knowledge, an effective means to deliver such products for long-term gene expression without disrupting normal cellular processes has not been described. In this study we have developed a retroviral strategy to effectively express gene products in the NK cell, whereby its effector functions of cytotoxicity and cytokine production remain intact. Using an EBV/retroviral hybrid vector PINCO, we demonstrate infection of human peripheral blood NK cells with simultaneous expression of a marker for infection - the enhanced green fluorescent protein (EGFP) - along with various genes of interest. This technique results in successful infection of the CD56dim NK population that predominates among human peripheral blood NK and is the effector of antibody-dependent cellular cytotoxicity (ADCC) and natural killing. In addition, we demonstrate infection of the CD56bright NK subset as well as the NK-92 and NK-L cell lines. Finally, we modify PINCO to express a cytoplasmically truncated murine CD8 molecule in place of GFP. The resulting vector enables us to transduce NK cells with multiple genes of interest simultaneously and provides an alternative purification method to FACS by using magnetic beads. In summary, we have devised an efficient and highly reproducible methodology for the targeted delivery of gene products to human NK cells that should now provide opportunities to dissect the molecular processes critical to normal NK cell physiology and to genetically manipulate NK cell populations prior to their administration in cancer therapy.

scholarly journals Response of resting human peripheral blood natural killer cells to interleukin 2.

1984 ◽  
Vol 160 (4) ◽  
pp. 1147-1169 ◽  
Author(s):  
G Trinchieri ◽  
M Matsumoto-Kobayashi ◽  
S C Clark ◽  
J Seehra ◽  
L London ◽  
...  
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Natural Killer ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Interleukin 2 ◽  
Cell Activity ◽  
Experimental Conditions ◽  
Ifn Gamma

The present study shows that recombinant interleukin 2 (IL-2) purified to homogeneity induces a rapid and potent enhancement of spontaneous cytotoxicity of human peripheral blood lymphocytes. The cells mediating cytotoxicity after 18-h treatment with IL-2 have surface markers of natural killer (NK) cells and are generated from the peripheral blood subset containing spontaneous cytotoxic cells. A parallel production of gamma interferon (IFN-gamma) is induced by recombinant IL-2 (rIL-2), and NK cells appear to be the major producer cells, whereas T cells are unable to produce IFN-gamma under these experimental conditions. However, the kinetics of the enhancement of cytotoxicity are faster than those of IFN-gamma production, and monoclonal anti-IFN-gamma antibodies do not suppress this effect, making it unlikely that the IFN-gamma produced is responsible for the enhancement. The enhancement of NK cell activity induced by rIL-2 precedes any proliferative response of the lymphocytes, which is instead observed in longer-term cultures of both NK and T cells.

scholarly journals Human peripheral blood DNAM-1neg NK cells are a terminally differentiated subset with limited effector functions

2019 ◽  
Vol 3 (11) ◽  
pp. 1681-1694 ◽  
Author(s):  
Kimberley A. Stannard ◽  
Sébastien Lemoine ◽  
Nigel J. Waterhouse ◽  
Frank Vari ◽  
Lucienne Chatenoud ◽  
...  
Keyword(s):  
Nk Cells ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Cell Subset ◽  
B Cell Lymphoma ◽  
Gene Signature ◽  
Interferon Γ ◽  
Specific Gene

Abstract Natural killer (NK) cells are a heterogeneous population of innate lymphocytes whose potent anticancer properties make them ideal candidates for cellular therapeutic application. However, our lack of understanding of the role of NK cell diversity in antitumor responses has hindered advances in this area. In this study, we describe a new CD56dim NK cell subset characterized by the lack of expression of DNAX accessory molecule-1 (DNAM-1). Compared with CD56bright and CD56dimDNAM-1pos NK cell subsets, CD56dimDNAM-1neg NK cells displayed reduced motility, poor proliferation, lower production of interferon-γ, and limited killing capacities. Soluble factors secreted by CD56dimDNAM-1neg NK cells impaired CD56dimDNAM-1pos NK cell–mediated killing, indicating a potential inhibitory role for the CD56dimDNAM-1neg NK cell subset. Transcriptome analysis revealed that CD56dimDNAM-1neg NK cells constitute a new mature NK cell subset with a specific gene signature. Upon in vitro cytokine stimulation, CD56dimDNAM-1neg NK cells were found to differentiate from CD56dimDNAM-1pos NK cells. Finally, we report a dysregulation of NK cell subsets in the blood of patients diagnosed with Hodgkin lymphoma and diffuse large B-cell lymphoma, characterized by decreased CD56dimDNAM-1pos/CD56dimDNAM-1neg NK cell ratios and reduced cytotoxic activity of CD56dimDNAM-1pos NK cells. Altogether, our data offer a better understanding of human peripheral blood NK cell populations and have important clinical implications for the design of NK cell–targeting therapies.

scholarly journals Identification and characterization of a pore-forming protein of human peripheral blood natural killer cells.

1986 ◽  
Vol 164 (6) ◽  
pp. 2061-2076 ◽  
Author(s):  
C C Liu ◽  
B Perussia ◽  
Z A Cohn ◽  
J D Young
Keyword(s):  
Nk Cells ◽  
Peripheral Blood ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Interleukin 2 ◽  
Calcium Ionophore ◽  
Internal Diameter ◽  
Ionophore A23187 ◽  
Cell Surface Antigens ◽  
Planar Bilayers

We show here that human peripheral blood NK cells contain a pore-forming protein (PFP) with an Mr of 70,000-72,000 that assembles structural lesions (with an average internal diameter of 150-170 A) and forms functional channels. The PFP was isolated by affinity chromatography from human NK cells, using a specific anti-C9 antiserum as the immunoadsorbent. The NK cells were isolated from PBL by positive or negative selection by indirect rosetting using a panel of monoclonal antibodies directed against different NK and T cell surface antigens. PFP was identified in NK cells freshly isolated and isolated from cultured PBL, both stimulated with interleukin 2, but not in NK cell-depleted lymphocytes. In planar bilayers, the channels formed by the NK cell-derived PFP are highly voltage resistant, with most channels persisting in the open state once they have inserted into the bilayer. The unit conductances of these channels range 0.3-1 nS in 0.1 M NaCl. The channels show poor selectivity for monovalent and divalent ions. The PFP is also released from human NK cells stimulated with the calcium ionophore A23187, suggesting that this protein, like the one produced by murine CTL lines, may be similarly secreted during cell-mediated killing. Its identification in primary human NK cell cultures indicates that this protein may play an active role in NK cell-mediated killing.

scholarly journals Acupuncture Regulates Leukocyte Subpopulations in Human Peripheral Blood

2007 ◽  
Vol 4 (4) ◽  
pp. 447-453 ◽  
Author(s):  
Nobuo Yamaguchi ◽  
Takashi Takahashi ◽  
Masahiro Sakuma ◽  
Toshiroh Sugita ◽  
Kumiko Uchikawa ◽  
...  
Keyword(s):  
Immune System ◽  
Peripheral Blood ◽  
Acupuncture Treatment ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Cell Activity ◽  
Humoral And Cellular Immunity ◽  
Cell Counts ◽  
Before And After ◽  
Ifn Γ

Acupuncture has recently been attracting more and more people throughout the world as an alternative treatment, however little is known about its physiological activities (i.e. immune system). We examined acupuncture both quantitatively and qualitatively by measuring CD-positive cell counts and cytokine expression levels in the blood, to determine the activity of T cells, B cells, macrophages and natural killer (NK) cells. Fifteen milliliters of peripheral blood obtained from 17 healthy volunteers aged 21–51 years, were analyzed using flow cytometry before and after acupuncture treatment. There was a statistically significant increase in the number of CD2+, CD4+, CD8+, CD11b+, CD16+, CD19+, CD56+cells as well as IL-4, IL-1β and IFN-γ levels in the cells after acupuncture stimulation of meridian points. These observations indicate that acupuncture may regulate the immune system and promote the activities of humoral and cellular immunity as well as NK cell activity. In this article, we discussed how acupuncture regulated leukocyte numbers and functions since they are considered to be potential indicators for evaluating complementary and alternative medicine.

scholarly journals Activation of natural killer cells via the p75 interleukin 2 receptor.

1989 ◽  
Vol 170 (1) ◽  
pp. 291-296 ◽  
Author(s):  
J H Phillips ◽  
T Takeshita ◽  
K Sugamura ◽  
L L Lanier
Keyword(s):  
Natural Killer Cells ◽  
Nk Cells ◽  
B Lymphocytes ◽  
Peripheral Blood ◽  
Cell Activation ◽  
Nk Cell ◽  
Human Peripheral Blood ◽  
Interleukin 2 ◽  
Maximal Inhibition ◽  
Activation Antigen

The IL-2R is composed of at least two subunits, the p55 (CD25/Tac) and p75 glycoproteins. The p75 IL-2R is expressed preferentially on resting human peripheral blood NK cells, but is not detected on substantial proportions of T and B lymphocytes, monocytes, or granulocytes. Anti-p75 IL-2R mAb substantially inhibits the early events associated with NK cell activation by IL-2, including inhibition of cytotoxic activity and induction of the CD69 early activation antigen. While anti-p55 IL-2R mAb alone failed to substantially inhibit the initial events of IL-2 stimulation, maximal inhibition of IL-2-induced cytotoxicity and proliferation was achieved by combining both anti-p55 IL-2R and anti-p75 IL-2R. Collectively, results from the present studies directly implicate the p75 IL-2R as the structure predominantly responsible for IL-2 activation of NK cells.

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