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

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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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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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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Differential requirement for the transcription factor PU.1 in the generation of natural killer cells versus B and T cells

Blood ◽

10.1182/blood.v97.9.2625 ◽

2001 ◽

Vol 97 (9) ◽

pp. 2625-2632 ◽

Cited By ~ 96

Author(s):

Francesco Colucci ◽

Sandrine I. Samson ◽

Rodney P. DeKoter ◽

Olivier Lantz ◽

Harinder Singh ◽

...

Keyword(s):

T Cells ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Fetal Liver ◽

Myeloid Cell ◽

Cell Development ◽

Stringent Requirement ◽

Nk Cell Differentiation ◽

Ets Family

Abstract PU.1 is a member of the Ets family of transcription factors required for the development of various lymphoid and myeloid cell lineages, but its role in natural killer (NK) cell development is not known. The study shows that PU.1 is expressed in NK cells and that, on cell transfer into alymphoid Rag2/γc−/−mice, hematopoietic progenitors of PU.1−/−fetal liver cells could generate functional NK cells but not B or T cells. Nevertheless, the numbers of bone marrow NK cell precursors and splenic mature NK cells were reduced compared to controls. Moreover,PU.1−/− NK cells displayed reduced expression of the receptors for stem cell factor and interleukin (IL)-7, suggesting a nonredundant role for PU.1 in regulating the expression of these cytokine receptor genes during NK cell development.PU.1−/− NK cells also showed defective expression of inhibitory and activating members of the Ly49 family and failed to proliferate in response to IL-2 and IL-12. Thus, despite the less stringent requirement for PU.1 in NK cell development compared to B and T cells, PU.1 regulates NK cell differentiation and homeostasis.

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Evidence for discrete stages of human natural killer cell differentiation in vivo

Journal of Experimental Medicine ◽

10.1084/jem.20052507 ◽

2006 ◽

Vol 203 (4) ◽

pp. 1033-1043 ◽

Cited By ~ 280

Author(s):

Aharon G. Freud ◽

Akihiko Yokohama ◽

Brian Becknell ◽

Melissa T. Lee ◽

Hsiaoyin C. Mao ◽

...

Keyword(s):

Cell Differentiation ◽

Natural Killer ◽

Nk Cell ◽

Killer Cell ◽

Intermediate Stage ◽

Cell Surface Expression ◽

Lymphoid Tissues ◽

Human Natural Killer Cell ◽

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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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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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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Characterization of the defective interaction between a subset of natural killer cells and dendritic cells in HIV-1 infection

Journal of Experimental Medicine ◽

10.1084/jem.20060894 ◽

2006 ◽

Vol 203 (10) ◽

pp. 2339-2350 ◽

Cited By ~ 129

Author(s):

Domenico Mavilio ◽

Gabriella Lombardo ◽

Audrey Kinter ◽

Manuela Fogli ◽

Andrea La Sala ◽

...

Keyword(s):

Dendritic Cells ◽

Nk Cells ◽

Natural Killer ◽

Nk Cell ◽

Cell Subset ◽

Interferon Γ ◽

And Function ◽

Hiv 1

In this study, we demonstrate that the in vitro interactions between a CD56neg/CD16pos (CD56neg) subset of natural killer (NK) cells and autologous dendritic cells (DCs) from HIV-1–infected viremic but not aviremic individuals are markedly impaired and likely interfere with the development of an effective immune response. Among the defective interactions are abnormalities in the process of reciprocal NK–DC activation and maturation as well as a defect in the NK cell–mediated editing or elimination of immature DCs (iDCs). Notably, the lysis of mature DCs (mDCs) by autologous NK cells was highly impaired even after the complete masking of major histocompatibility complex I molecules, suggesting that the defective elimination of autologous iDCs is at the level of activating NK cell receptors. In this regard, the markedly impaired expression/secretion and function of NKp30 and TNF-related apoptosis-inducing ligand, particularly among the CD56neg NK cell subset, largely accounts for the highly defective NK cell–mediated lysis of autologous iDCs. Moreover, mDCs generated from HIV-1 viremic but not aviremic patients are substantially impaired in their ability to secrete interleukin (IL)-10 and -12 and to prime the proliferation of neighboring autologous NK cells, which, in turn, fail to secrete adequate amounts of interferon-γ.

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