THE ROLES OF MICRORNA IN HUMAN NATURAL KILLER (NK) CELL DIFFERENTIATION

Human natural killer (NK) cells originate from CD34(+) hematopoietic progenitor cells, but the discrete stages of NK cell differentiation in vivo have not been elucidated. We identify and functionally characterize, from human lymph nodes and tonsils, four NK cell developmental intermediates spanning the continuum of differentiation from a CD34(+) NK cell progenitor to a functionally mature NK cell. Analyses of each intermediate stage for CD34, CD117, and CD94 cell surface expression, lineage differentiation potentials, capacity for cytokine production and natural cytotoxicity, and ETS-1, GATA-3, and T-BET expression provide evidence for a new model of human NK cell differentiation in secondary lymphoid tissues.

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Coordinated acquisition of inhibitory and activating receptors and functional properties by developing human natural killer cells

Blood ◽

10.1182/blood-2006-04-020198 ◽

2006 ◽

Vol 108 (12) ◽

pp. 3824-3833 ◽

Cited By ~ 107

Author(s):

Bartosz Grzywacz ◽

Nandini Kataria ◽

Magdalena Sikora ◽

Robert A. Oostendorp ◽

Elaine A. Dzierzak ◽

...

Keyword(s):

Cell Differentiation ◽

Natural Killer ◽

Cell Line ◽

Nk Cell ◽

Fetal Liver ◽

Interferon Γ ◽

Intermediate Phenotype ◽

Interleukin 7 ◽

Nk Cell Differentiation ◽

And Function

AbstractThe stages of human natural killer (NK) cell differentiation are not well established. Culturing CD34+ progenitors with interleukin 7 (IL-7), IL-15, stem cell factor (SCF), FLT-3L, and murine fetal liver cell line (EL08.1D2), we identified 2 nonoverlapping subsets of differentiating CD56+ cells based on CD117 and CD94 (CD117highCD94– and CD117low/–CD94+ cells). Both populations expressed CD161 and NKp44, but differed with respect to NKp30, NKp46, NKG2A, NKG2C, NKG2D, CD8, CD16, and KIR. Only the CD117low/– CD94+ population displayed cytotoxicity and interferon-γ production. Both populations arose from a single CD34+CD38– Lin– cell and their percentages changed over time in a reciprocal fashion, with CD117highCD94– cells predominating early and decreasing due to an increase of the CD117low/–CD94+ population. These 2 subsets represent distinct stages of NKcell differentiation, since purified CD117high CD94– cells give rise to CD117low/–CD94+ cells. The stromal cell line (EL08.1D2) facilitated the transition from CD117highCD94– to CD117low/–CD94+ via an intermediate phenotype (CD117lowCD94low/–). EL08.1D2 also maintained the mature phenotype, preventing the reversion of CD117low/–CD94+ cells to the intermediate (CD117lowCD94low/–) phenotype. An analogous population of CD56+CD117highCD94– cells was found in cord blood. The identified stages of NK-cell differentiation provide evidence for coordinated acquisition of HLA-specific inhibitory receptors (ie, CD94/NKG2A) and function in developing human NK cells.

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Cytomegalovirus-Driven Adaption of Natural Killer Cells in NKG2Cnull Human Immunodeficiency Virus-Infected Individuals

Viruses ◽

10.3390/v11030239 ◽

2019 ◽

Vol 11 (3) ◽

pp. 239 ◽

Cited By ~ 1

Author(s):

Emilie M. Comeau ◽

Kayla A. Holder ◽

Neva J. Fudge ◽

Michael D. Grant

Keyword(s):

Human Immunodeficiency Virus ◽

Cell Differentiation ◽

Hiv Infection ◽

Nk Cells ◽

Natural Killer ◽

Cytokine Production ◽

Zinc Finger Protein ◽

Nk Cell ◽

Immunodeficiency Virus ◽

Nk Cell Differentiation

Expansion of natural killer (NK) cells expressing NKG2C occurs following human cytomegalovirus (HCMV) infection and is amplified by human immunodeficiency virus (HIV) co-infection. These NKG2C-expressing NK cells demonstrate enhanced CD16-dependent cytokine production and downregulate FcεRIγ and promyelocytic leukemia zinc finger protein (PLZF). Lacking NKG2C diminishes resistance to HIV infection, but whether this affects NK cell acquisition of superior antibody-dependent function is unclear. Therefore, our objective was to investigate whether HCMV-driven NK cell differentiation is impaired in NKG2Cnull HIV-infected individuals. Phenotypic (CD2, CD16, CD57, NKG2A, FcεRIγ, and PLZF expression) and functional (cytokine induction and cytotoxicity) properties were compared between HIV–infected NKG2Cnull and NKG2C-expressing groups. Cytokine production was compared following stimulation through natural cytotoxicity receptors or through CD16. Cytotoxicity was measured by anti-CD16-redirected lysis and by classical antibody-dependent cell-mediated cytotoxicity (ADCC) against anti-class I human leukocyte antigen (HLA) antibody-coated cells. Our data indicate highly similar HCMV-driven NK cell differentiation in HIV infection with or without NKG2C. While the fraction of mature (CD57pos) NK cells expressing CD2 (p = 0.009) or co-expressing CD2 and CD16 (p = 0.03) was significantly higher in NKG2Cnull HIV-infected individuals, there were no significant differences in NKG2A, FcεRIγ, or PLZF expression. The general phenotypic and functional equivalency observed suggests NKG2C-independent routes of HCMV-driven NK cell differentiation, which may involve increased CD2 expression.

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Combined deficiency in IκBα and IκBϵ reveals a critical window of NF-κB activity in natural killer cell differentiation

Blood ◽

10.1182/blood-2003-08-2975 ◽

2004 ◽

Vol 103 (12) ◽

pp. 4573-4580 ◽

Cited By ~ 24

Author(s):

Sandrine I. Samson ◽

Sylvie Mémet ◽

Christian A. J. Vosshenrich ◽

Francesco Colucci ◽

Odile Richard ◽

...

Keyword(s):

Cell Differentiation ◽

Natural Killer ◽

Nk Cell ◽

Killer Cell ◽

Nuclear Factor Κb ◽

Interferon Γ ◽

Cell Development ◽

Critical Window ◽

Proliferation And Apoptosis ◽

Nk Cell Differentiation

Abstract Nuclear factor κB (NF-κB) transcription factors are key regulators of immune, inflammatory, and acute-phase responses and are also implicated in the control of cell proliferation and apoptosis. While perturbations in NF-κB activity impact strongly on B- and T-cell development, little is known about the role for NF-κB in natural killer (NK) cell differentiation. Inhibitors of NF-κB (IκBs) act to restrain NF-κB activation. We analyzed the cell-intrinsic effects of deficiencies in 2 IκB members (IκBα and IκBϵ) on NK cell differentiation. Neither IκBα nor IκBϵ deficiency had major effects on NK cell generation, while their combined absence led to NF-κB hyperactivation, resulting in reduced NK cell numbers, incomplete NK cell maturation, and defective interferon γ (IFN-γ) production. Complementary analysis of transgenic mice expressing an NF-κB-responsive reporter gene showed increased NF-κB activity at the stage of NK cell development corresponding to the partial block observed in IκBα × IκBϵ-deficient mice. These results define a critical window in NK cell development in which NF-κB levels may be tightly controlled. (Blood. 2004;103:4573-4580)

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Monocytes control natural killer cell differentiation to effector phenotypes

Blood ◽

10.1182/blood-2010-10-312264 ◽

2011 ◽

Vol 117 (17) ◽

pp. 4511-4518 ◽

Cited By ~ 59

Author(s):

Katrina Soderquest ◽

Nick Powell ◽

Carmelo Luci ◽

Nico van Rooijen ◽

Andrés Hidalgo ◽

...

Keyword(s):

Cell Differentiation ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Killer Cell ◽

In Vivo Model ◽

Dependent Manner ◽

Nk Cell Differentiation

Abstract Natural killer (NK) cells play a major role in immunologic surveillance of cancer. Whether NK-cell subsets have specific roles during antitumor responses and what the signals are that drive their terminal maturation remain unclear. Using an in vivo model of tumor immunity, we show here that CD11bhiCD27low NK cells migrate to the tumor site to reject major histocompatibility complex class I negative tumors, a response that is severely impaired in Txb21−/− mice. The phenotypical analysis of Txb21-deficient mice shows that, in the absence of Txb21, NK-cell differentiation is arrested specifically at the CD11bhiCD27hi stage, resulting in the complete absence of terminally differentiated CD11bhiCD27low NK cells. Adoptive transfer experiments and radiation bone marrow chimera reveal that a Txb21+/+ environment rescues the CD11bhiCD27hi to CD11bhiCD27low transition of Txb21−/− NK cells. Furthermore, in vivo depletion of myeloid cells and in vitro coculture experiments demonstrate that spleen monocytes mediate the terminal differentiation of peripheral NK cells in a Txb21- and IL-15Rα–dependent manner. Together, these data reveal a novel, unrecognized role for Txb21 expression in monocytes in promoting NK-cell development and help appreciate how various NK-cell subsets are generated and participate in antitumor immunity.

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Ly49E expression points toward overlapping, but distinct, natural killer (NK) cell differentiation kinetics and potential of fetal versus adult lymphoid progenitors

Journal of Leukocyte Biology ◽

10.1189/jlb.0902443 ◽

2003 ◽

Vol 73 (6) ◽

pp. 731-738 ◽

Cited By ~ 17

Author(s):

Frederik Stevenaert ◽

Katrien Van Beneden ◽

An De Creus ◽

Veronique Debacker ◽

Jean Plum ◽

...

Keyword(s):

Cell Differentiation ◽

Natural Killer ◽

Nk Cell ◽

Lymphoid Progenitors ◽

Nk Cell Differentiation

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Definition of a natural killer NKR-P1A+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12.

Journal of Experimental Medicine ◽

10.1084/jem.184.5.1845 ◽

1996 ◽

Vol 184 (5) ◽

pp. 1845-1856 ◽

Cited By ~ 83

Author(s):

I M Bennett ◽

O Zatsepina ◽

L Zamai ◽

L Azzoni ◽

T Mikheeva ◽

...

Keyword(s):

Cell Differentiation ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Cell Subset ◽

Interleukin 12 ◽

Ifn Gamma ◽

Differentiation Antigens ◽

Nk Cell Differentiation

Human natural killer (NK) cell differentiation from immature lineage negative (Lin-) umbilical cord blood cells was examined in vitro. Cells expressing differentiation antigens of mature NK cells (CD56, CD16, CD2, CD8, NKR-P1A) were generated from Lin- cells cultured with interleukin (IL)-2 and a murine bone marrow stromal cell line expressing the human membrane-bound form of stem cell factor. Two subsets of NK cells were identified in these cultures: one expressed both NKR-P1A and CD56 and, in variable proportions, all other NK cell differentiation antigens; the second subset expressed only NKR-P1A and, unlike the former, was not cytotoxic. Neither subset expressed interferon (IFN)-gamma mRNA even after stimulation with phorbol di-ester and Ca2+ ionophore, but both expressed tumor necrosis factor alpha mRNA and the cytotoxic granule-associated proteins TIA-1, perforin, and serine esterase-1. After 10-d culture with IL-2, IL-12, and irradiated B lymphoblastoid cells, approximately 45% of the NKR-P1A+/ CD56- cells became CD56+, and the same cultures contained cells capable of cytotoxicity and of IFN-gamma production. These results indicate that NKR-P1A expression in the absence of other NK cell markers defines an intermediate, functionally immature stage of NK cell differentiation, and that effector functions develop in these cells, concomitantly with CD56 expression, in the presence of IL-12. These cells likely represent the counterpart of a CD3-/NKR-P1A+/ CD56-/CD16- cell subset that, as shown here, is present both in adult and neonatal circulating lymphocytes.

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A Novel Human CD34(+) Subset That Constitutively Expresses the High Affinity Interleukin-2 Receptor Traffics to Lymph Nodes and Differentiates into CD56Bright Natural Killer Cells.

Blood ◽

10.1182/blood.v104.11.314.314 ◽

2004 ◽

Vol 104 (11) ◽

pp. 314-314

Author(s):

Aharon G. Freud ◽

Brian Becknell ◽

Sameek Roychowdhury ◽

Hsiaoyin C. Mao ◽

Amy K. Ferketich ◽

...

Keyword(s):

Cell Differentiation ◽

T Cell ◽

Lymph Nodes ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Interleukin 2 ◽

Constitutive Expression ◽

High Affinity ◽

Nk Cell Differentiation

Abstract In adult humans, T cells differentiate in the thymus and B cells develop in the bone marrow, but the site(s) of natural killer (NK) cell differentiation are unclear. Here we describe, for the first time, a unique CD34(+) population found in human lymph nodes (LN) that differentiates into NK cells. CD56bright NK cells represent <10% of NK cells in peripheral blood (PB) yet predominate in LN where they can compete for endogenous T cell-derived IL-2 during immune activation due to their unique expression of functional high affinity (HA) interleukin (IL)-2 receptors (IL-2R). We hypothesized that a subset of CD34(+) hematopoietic precursor cells (HPC) might also express functional HA IL-2R and potentially differentiate into CD56bright NK cells via activation with low dose IL-2. We first identified a novel human CD34dimCD45RA(+) HPC in PB with constitutive expression of the HA IL-2R. When cultured in picomolar concentrations of IL-2 that selectively saturate the HA IL-2R, these cells give rise to CD56bright NK cells, and this effect is blocked when IL-2 cannot bind to its HA receptor. This unique CD34(+) population expresses IL-2Rα, CD2, CD7, c-kit, L-selectin, and NKR-P1A, all of which are also expressed by CD56bright NK cells. Unique among total PB CD34(+) cells, this novel population displays high integrin α4β7 expression. This attribute, in addition to its high L-selectin expression, suggested that these cells may traffic to LN where their progeny, CD56bright NK cells, represent the major NK subset. Indeed we found a distinct CD34dimCD45RA(+)α4β7bright population that resides in the T cell rich regions of human LN, and when stimulated in vitro with 10 pM IL-2, this cell gives rise to CD56bright NK cells. This novel population represents only ~6% of all PB CD34(+) HPC yet is the major if not exclusive CD34(+) subset in LN. While murine studies strongly support the notion that most if not all NK cells require IL-15 for their development, these new human data suggest a model for development of a minor human NK subset, the CD56bright NK cells, whereby CD34dimCD45RA(+)α4β7bright HPC constitutively expressing the HA IL-2R traffic to peripheral LN where endogenous T cell-derived IL-2 can drive CD56bright NK cell differentiation in vivo.

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Antibody-Dependent Natural Killer Cell Activation After Ebola Vaccination

The Journal of Infectious Diseases ◽

10.1093/infdis/jiz657 ◽

2019 ◽

Cited By ~ 5

Author(s):

Helen R Wagstaffe ◽

Elizabeth A Clutterbuck ◽

Viki Bockstal ◽

Jeroen N Stoop ◽

Kerstin Luhn ◽

...

Keyword(s):

Cell Differentiation ◽

Nk Cells ◽

Natural Killer ◽

Cell Activation ◽

Ebola Virus ◽

Nk Cell ◽

Killer Cell ◽

Antibody Concentration ◽

Vaccine Regimen ◽

Nk Cell Differentiation

Abstract Background Antibody Fc-mediated functions, such as antibody-dependent cellular cytotoxicity, contribute to vaccine-induced protection against viral infections. Fc-mediated function of anti-Ebola glycoprotein (GP) antibodies suggest that Fc-dependent activation of effector cells, including natural killer (NK) cells, could play a role in vaccination against Ebola virus disease. Methods We analyzed the effect on primary human NK cell activation of anti-Ebola GP antibody in the serum of United Kingdom–based volunteers vaccinated with the novel 2-dose heterologous adenovirus type 26.ZEBOV, modified vaccinia Ankara–BN-Filo vaccine regimen. Results We demonstrate primary human NK cell CD107a and interferon γ expression, combined with down-regulation of CD16, in response to recombinant Ebola virus GP and post-vaccine dose 1 and dose 2 serum samples. These responses varied significantly with vaccine regimen, and NK cell activation was found to correlate with anti-GP antibody concentration. We also reveal an impact of NK cell differentiation phenotype on antibody-dependent NK cell activation, with highly differentiated CD56dimCD57+ NK cells being the most responsive. Conclusions These findings highlight the dual importance of vaccine-induced antibody concentration and NK cell differentiation status in promoting Fc-mediated activation of NK cells after vaccination, raising a potential role for antibody-mediated NK cell activation in vaccine-induced immune responses.

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The receptor tyrosine kinase c-kit provides a critical signal for survival, expansion, and maturation of mouse natural killer cells

Blood ◽

10.1182/blood.v95.3.984.003k40_984_991 ◽

2000 ◽

Vol 95 (3) ◽

pp. 984-991 ◽

Cited By ~ 56

Author(s):

Francesco Colucci ◽

James P. Di Santo

Keyword(s):

Stem Cell ◽

Cell Differentiation ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Fetal Liver ◽

Lower Percentage ◽

Nk Cell Differentiation

Fetal liver kinase ligands (flk2L/flt3L) and stem cell factor (SCF) have been shown to promote natural killer (NK) cell differentiation from hematopoietic stem cell (HSC) precursors in vitro. However, the contribution of signaling through the receptors for these growth factors for in vivo NK cell development remains ill-defined. We have analyzed the role of the SCF receptor c-kit in NK cell differentiation by reconstituting NK-deficient mice with fetal liver (FL) HSCs of c-kit−/− (W/W) mice. Although c-kit−/−NK cells were generated inW/W chimeras, they were reduced in number, contained a lower percentage of CD45R (B220)+ cells, and were poorly cytolytic. In vitro experiments showed that generation of NK cells from FL precursors was reduced in the absence of c-kit signaling and that SCF promoted the survival of peripheral c-kit+ NK cells. We conclude that c-kit/SCF interactions in vivo are dispensable for the commitment of HSC to the NK lineage, but they provide essential signals for generating normal numbers of fully mature NK cells.

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