scholarly journals Tolerance of NK cells encountering their viral ligand during development

2008 ◽  
Vol 205 (8) ◽  
pp. 1819-1828 ◽  
Author(s):  
Joseph C. Sun ◽  
Lewis L. Lanier
Keyword(s):  
B Cells ◽  
Nk Cells ◽  
Adaptor Proteins ◽  
Interferon Γ ◽  
Target Cells ◽  
T And B Cells ◽  
Mcmv Infection ◽  
Retroviral Transduction ◽  
Activating Receptors ◽  
Activation Motif

During development, T and B cells encountering their cognate ligands via antigen-specific receptors are deleted or rendered anergic. Like T and B cells, natural killer (NK) cells express certain receptors, such as Ly49H, associated with immunoreceptor tyrosine-based activation motif–bearing adaptor proteins that transmit activating signals through Syk family kinases. Ly49H binds with high affinity to a mouse cytomegalovirus (MCMV)–encoded glycoprotein, m157, but does not recognize self-antigens. For comparison with the behavior of immature T and B cells exposed to foreign antigens, we addressed the fate of Ly49H+ NK cells that encountered their viral ligand during development by retroviral transduction of bone marrow stem cells with m157. In chimeric mice expressing m157, we observed a reduction in Ly49H+ NK cells in multiple tissues and less Ly49H on the cell surface. NK cells exposed to m157 during development appeared less mature, produced less interferon γ when stimulated through Ly49H, and were unable to kill m157-bearing target cells. After MCMV infection, these NK cells were severely impaired in their ability to proliferate. Thus, if immature NK cells encounter ligands for their activating receptors, regulatory mechanisms exist to keep these cells in an unresponsive state.

scholarly journals Enhanced NK-cell development and function in BCAP-deficient mice

Blood ◽  
2008 ◽  
Vol 112 (1) ◽  
pp. 131-140 ◽  
Author(s):  
Alexander W. MacFarlane ◽  
Tetsuo Yamazaki ◽  
Min Fang ◽  
Luis J. Sigal ◽  
Tomohiro Kurosaki ◽  
...  
Keyword(s):  
B Cells ◽  
Nk Cells ◽  
Nk Cell ◽  
Interferon Γ ◽  
Term Survival ◽  
Final Maturation ◽  
Severe Impairment ◽  
And Function ◽  
Activation Motif ◽  
Deficient Mice

Abstract In B lymphocytes, the B-cell adaptor for phosphatidylinositol 3-kinase (BCAP) facilitates signaling from the antigen receptor. Mice lacking BCAP have a predominantly immature pool of B cells with impaired immune function and increased susceptibility to apoptosis. Unexpectedly, we have found that natural killer (NK) cells from BCAP-deficient mice are more mature, more long-lived, more resistant to apoptosis, and exhibit enhanced functional activity compared with NK cells from wild-type mice. Surprisingly, these effects are evident despite a severe impairment of the immunoreceptor tyrosine-based activation motif-mediated Akt signaling pathway. The seemingly paradoxical phenotype reveals inherent differences in the signals controlling the final maturation of B cells and NK cells, which depend on positive and negative signals, respectively. Both enhanced interferon-γ responses and augmented maturation of NK cells in BCAP-deficient mice are independent of available MHC class I ligands. Our data support a model in which blunting of BCAP-mediated activation signaling in developing NK cells promotes functionality, terminal maturation, and long-term survival.

scholarly journals Distinct MHC class I–dependent NK cell–activating receptors control cytomegalovirus infection in different mouse strains

2011 ◽  
Vol 208 (5) ◽  
pp. 1105-1117 ◽  
Author(s):  
Michał Pyzik ◽  
Benoit Charbonneau ◽  
Eve-Marie Gendron-Pontbriand ◽  
Marina Babić ◽  
Astrid Krmpotić ◽  
...  
Keyword(s):  
Nk Cells ◽  
Nk Cell ◽  
Interferon Γ ◽  
Mouse Strains ◽  
Common Mechanism ◽  
Mcmv Infection ◽  
Viral Loads ◽  
Nk Cell Receptors ◽  
Activating Receptors ◽  
Infected Cells

Recognition of mouse cytomegalovirus (MCMV)–infected cells by activating NK cell receptors was first described in the context of Ly49H, which confers resistance to C57BL/6 mice. We investigated the ability of other activating Ly49 receptors to recognize MCMV-infected cells in mice from various H-2 backgrounds. We observed that Ly49P1 from NOD/Ltj mice, Ly49L from BALB mice, and Ly49D2 from PWK/Pas mice respond to MCMV-infected cells in the context of H-2Dk and the viral protein m04/gp34. Recognition was also seen in the H-2d and/or H-2f contexts, depending on the Ly49 receptor examined, but never in H-2b. Furthermore, BALB.K (H-2k) mice showed reduced viral loads compared with their H-2d or H-2b congenic partners, a reduction which was dependent on interferon γ secretion by Ly49L+ NK cells early after infection. Adoptive transfer of Ly49L+, but not Ly49L−, NK cells significantly increased resistance against MCMV infection in neonate BALB.K mice. These results suggest that multiple activating Ly49 receptors participate in H-2–dependent recognition of MCMV infection, providing a common mechanism of NK cell–mediated resistance against viral infection.

scholarly journals Influenza A Virus Hemagglutinin and Other Pathogen Glycoprotein Interactions with NK Cell Natural Cytotoxicity Receptors NKp46, NKp44, and NKp30

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 156
Author(s):  
Jasmina M. Luczo ◽  
Sydney L. Ronzulli ◽  
Stephen M. Tompkins
Keyword(s):  
Influenza Virus ◽  
Nk Cells ◽  
Influenza A Virus ◽  
Influenza A ◽  
Nk Cell ◽  
Influenza Virus Infection ◽  
Target Cells ◽  
Natural Cytotoxicity ◽  
Activating Receptors ◽  
Natural Cytotoxicity Receptors

Natural killer (NK) cells are part of the innate immunity repertoire, and function in the recognition and destruction of tumorigenic and pathogen-infected cells. Engagement of NK cell activating receptors can lead to functional activation of NK cells, resulting in lysis of target cells. NK cell activating receptors specific for non-major histocompatibility complex ligands are NKp46, NKp44, NKp30, NKG2D, and CD16 (also known as FcγRIII). The natural cytotoxicity receptors (NCRs), NKp46, NKp44, and NKp30, have been implicated in functional activation of NK cells following influenza virus infection via binding with influenza virus hemagglutinin (HA). In this review we describe NK cell and influenza A virus biology, and the interactions of influenza A virus HA and other pathogen lectins with NK cell natural cytotoxicity receptors (NCRs). We review concepts which intersect viral immunology, traditional virology and glycobiology to provide insights into the interactions between influenza virus HA and the NCRs. Furthermore, we provide expert opinion on future directions that would provide insights into currently unanswered questions.

scholarly journals GSK-3α Inhibition in Drug-Resistant CML Cells Promotes Susceptibility to NK Cell-Mediated Lysis in an NKG2D- and NKp30-Dependent Manner

Cancers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1802
Author(s):  
Nayoung Kim ◽  
Mi Yeon Kim ◽  
Woo Seon Choi ◽  
Eunbi Yi ◽  
Hyo Jung Lee ◽  
...  
Keyword(s):  
Nk Cells ◽  
Nk Cell ◽  
Negative Regulator ◽  
Target Cells ◽  
Dependent Manner ◽  
Cell Functions ◽  
Cell Based Therapy ◽  
Activating Receptors ◽  
Gsk 3Β

Natural killer (NK) cells are innate cytotoxic lymphocytes that provide early protection against cancer. NK cell cytotoxicity against cancer cells is triggered by multiple activating receptors that recognize specific ligands expressed on target cells. We previously demonstrated that glycogen synthase kinase (GSK)-3β, but not GSK-3α, is a negative regulator of NK cell functions via diverse activating receptors, including NKG2D and NKp30. However, the role of GSK-3 isoforms in the regulation of specific ligands on target cells is poorly understood, which remains a challenge limiting GSK-3 targeting for NK cell-based therapy. Here, we demonstrate that GSK-3α rather than GSK-3β is the primary isoform restraining the expression of NKG2D ligands, particularly ULBP2/5/6, on tumor cells, thereby regulating their susceptibility to NK cells. GSK-3α also regulated the expression of the NKp30 ligand B7-H6, but not the DNAM-1 ligands PVR or nectin-2. This regulation occurred independently of BCR-ABL1 mutation that confers tyrosine kinase inhibitor (TKI) resistance. Mechanistically, an increase in PI3K/Akt signaling in concert with c-Myc was required for ligand upregulation in response to GSK-3α inhibition. Importantly, GSK-3α inhibition improved cancer surveillance by human NK cells in vivo. Collectively, our results highlight the distinct role of GSK-3 isoforms in the regulation of NK cell reactivity against target cells and suggest that GSK-3α modulation could be used to enhance tumor cell susceptibility to NK cells in an NKG2D- and NKp30-dependent manner.

scholarly journals A novel type of cell death of lymphocytes induced by a monoclonal antibody without participation of complement.

1995 ◽  
Vol 181 (6) ◽  
pp. 2007-2015 ◽  
Author(s):  
S Matsuoka ◽  
Y Asano ◽  
K Sano ◽  
H Kishimoto ◽  
I Yamashita ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Death ◽  
T Cell ◽  
B Cells ◽  
Interleukin 2 ◽  
Cytolytic Activity ◽  
Target Cells ◽  
T And B Cells ◽  
Fab Fragments ◽  
T Cell Clone

A monoclonal antibody, RE2, raised by immunizing a rat with cell lysate of a mouse T cell clone, was found to directly kill interleukin 2-dependent T cell clones without participation of serum complement. Fab fragments of RE2 had no cytolytic activity, while the cross-linking of Fab fragments with anti-rat immunoglobulin reconstituted the cytotoxicity. The cytotoxicity was temperature dependent: the antibody could kill target cells at 37 degrees C but not at 0 degrees C. Sodium azide, ethylenediaminetetraacetic acid, and forskolin did not affect the cytolytic activity of RE2, while the treatment of target cells with cytochalasin B and D completely blocked the activity. This suggested that the cell death involves a cytoskeleton-dependent active process. Giant holes on the cell membrane were formed within 5 minutes after the treatment with RE2, as observed by scanning electron microscopy. There was no indication of DNA fragmentation nor swelling of mitochondria during the cytolysis, suggesting that the cell death is neither apoptosis nor typical necrosis. The antibody also killed T cell lymphomas and T and B cell hybridomas only when these cells were preactivated with concanavalin A, lipopolysaccharide, or phorbol myristate acetate. Preactivated peripheral T and B cells were sensitive to the cytotoxicity of RE2, while resting T and B cells were insensitive. These results provide evidence for a novel pathway of cell death of activated lymphocytes by membrane excitation.

scholarly journals Lymphoid and myeloid differentiation of fetal liver CD34+lineage- cells in human thymic organ culture.

1994 ◽  
Vol 180 (1) ◽  
pp. 123-132 ◽  
Author(s):  
A Bárcena ◽  
A H Galy ◽  
J Punnonen ◽  
M O Muench ◽  
D Schols ◽  
...  
Keyword(s):  
T Cells ◽  
B Cells ◽  
Organ Culture ◽  
Nk Cells ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Myeloid Differentiation ◽  
Target Cells ◽  
Hematopoietic Progenitors ◽  
Fetal Thymus

In this article, we report that the human fetal thymus contains CD34bright cells (< 0.01% of total thymocytes) with a phenotype that resembles that of multipotent hematopoietic progenitors in the fetal bone marrow. CD34bright thymocytes were CD33-/dull and were negative for CD38, CD2, and CD5 as well as for the lineage markers CD3, CD4, and CD8 (T cells), CD19 and CD20 (B cells), CD56 (NK cells), glycophorin (erythrocytes), and CD14 (monocytes). In addition, total CD34+ lineage negative (lin-) thymocytes contained a low number of primitive myeloid progenitor cells, thus suggesting that the different hematopoietic lineages present in the thymus may be derived from primitive hematopoietic progenitor cells seeding the thymus. To investigate whether the thymus is permissive for the development of non-T cells, human fetal organ culture (FTOC) assays were performed by microinjecting sorted CD34+lin- fetal liver cells into fragments of HLA-mismatched fetal thymus. Sequential phenotypic analysis of the FTOC-derived progeny of CD34+lin- cells indicated that the differentiation into T cells was preceded by a wave of myeloid differentiation into CD14+CD11b+CD4dull cells. Donor-derived B cells (CD19+CD20+) were also generated, which produced immunoglobulins (IgG and IgM) when cultured under appropriate conditions, as well as functional CD56+CD3- NK cells, which efficiently killed K562 target cells in cytotoxicity assays. These results demonstrate that the microinjection of fetal liver hematopoietic progenitors into fetal thymic organ fragments results in multilineage differentiation in vitro.

scholarly journals MHC class I–deficient natural killer cells acquire a licensed phenotype after transfer into an MHC class I–sufficient environment

2010 ◽  
Vol 207 (10) ◽  
pp. 2073-2079 ◽  
Author(s):  
Julie M. Elliott ◽  
Joseph A. Wahle ◽  
Wayne M. Yokoyama
Keyword(s):  
Bone Marrow ◽  
Nk Cells ◽  
Natural Killer ◽  
Mhc Class I ◽  
Interferon Γ ◽  
Class I ◽  
Target Cells ◽  
Major Histocompatability Complex ◽  
Gain Of Function ◽  
Mhc Class I Molecule

In MHC class I–deficient hosts, natural killer (NK) cells are hyporesponsive to cross-linking of activation receptors. Functional competence requires engagement of a self–major histocompatability complex (MHC) class I–specific inhibitory receptor, a process referred to as “licensing.” We previously suggested that licensing is developmentally determined in the bone marrow. In this study, we find that unlicensed mature MHC class I–deficient splenic NK cells show gain-of-function and acquire a licensed phenotype after adoptive transfer into wild-type (WT) hosts. Transferred NK cells produce WT levels of interferon-γ after engagement of multiple activation receptors, and degranulate at levels equivalent to WT NK cells upon coincubation with target cells. Only NK cells expressing an inhibitory Ly49 receptor specific for a cognate host MHC class I molecule show this gain-of-function. Therefore, these findings, which may be relevant to clinical bone marrow transplantation, suggest that neither exposure to MHC class I ligands during NK development in the BM nor endogenous MHC class I expression by NK cells themselves is absolutely required for licensing.

Molecular analysis of the methylprednisolone-mediated inhibition of NK-cell function: evidence for different susceptibility of IL-2– versus IL-15–activated NK cells

Blood ◽  
2007 ◽  
Vol 109 (9) ◽  
pp. 3767-3775 ◽  
Author(s):  
Laura Chiossone ◽  
Chiara Vitale ◽  
Francesca Cottalasso ◽  
Sara Moretti ◽  
Bruno Azzarone ◽  
...  
Keyword(s):  
Nk Cells ◽  
Cell Function ◽  
Nk Cell ◽  
Surface Expression ◽  
Steroid Treatment ◽  
Cross Linking ◽  
Target Cells ◽  
Activating Receptors ◽  
And Function ◽  
Nk Cytotoxicity

Abstract Steroids have been shown to inhibit the function of fresh or IL-2–activated natural killer (NK) cells. Since IL-15 plays a key role in NK-cell development and function, we comparatively analyzed the effects of methylprednisolone on IL-2– or IL-15–cultured NK cells. Methylprednisolone inhibited the surface expression of the major activating receptors NKp30 and NKp44 in both conditions, whereas NK-cell proliferation and survival were sharply impaired only in IL-2–cultured NK cells. Accordingly, methylprednisolone inhibited Tyr phosphorylation of STAT1, STAT3, and STAT5 in IL-2–cultured NK cells but only marginally in IL-15–cultured NK cells, whereas JAK3 was inhibited under both conditions. Also, the NK cytotoxicity was similarly impaired in IL-2– or IL-15–cultured NK cells. This effect strictly correlated with the inhibition of ERK1/2 Tyr phosphorylation, perforin release, and cytotoxicity in a redirected killing assay against the FcRγ+ P815 target cells upon cross-linking of NKp46, NKG2D, or 2B4 receptors. In contrast, in the case of CD16, inhibition of ERK1/2 Tyr phosphorylation, perforin release, and cytotoxicity were not impaired. Our study suggests a different ability of IL-15–cultured NK cells to survive to steroid treatment, thus offering interesting clues for a correct NK-cell cytokine conditioning in adoptive immunotherapy.

Arteriolar reactivity in lymphocyte-deficient mice

2011 ◽  
Vol 301 (4) ◽  
pp. H1276-H1285 ◽  
Author(s):  
Sean Leonard ◽  
B. Anne Croy ◽  
Coral L. Murrant
Keyword(s):  
B Cells ◽  
Nk Cells ◽  
Ang Ii ◽  
T And B Cells ◽  
Third Order ◽  
Immunodeficient Mice ◽  
Circulatory Control ◽  
Deficient Mice ◽  
Cell Cytokine Production

Mounting evidence suggests that lymphocytes have the capacity to contribute to the regulation of systemic circulatory control. We postulated that T and natural killer (NK) cells could modify basal microvascular activity under physiologically normal conditions. In situ intravital microscopy of mouse cremaster vasculature was used to evaluate arteriolar reactivities to the vasoconstrictors angiotensin II (ANG II) and phenylephrine (Phe) and the vasodilators acetylcholine (ACh) and adenosine (Ado) in normal [+/+; wild type (WT)] and genetically immunodeficient (T−B−NK+ or T−B−\NK−) C57BL/6 and BALB/c mice, strain backgrounds with differentially polarized T cell cytokine production. Immunodeficient mice tended to have smaller baseline and maximal diameters of third-order cremaster arterioles than their congenic WT partners. In C57BL/6, baseline diameters were similar in T-B− mice without or with NK cells; in BALB/c, baseline diameters were larger in T-B-NK− mice than in T−B−NK+ mice. Thus, at baseline, lymphocytes tended to promote vasodilation, except BALB/c NK cells, which mediated mild vasoconstriction. The presence of NK cells suppressed dilations to Ado in both strains, to ACh in the C57BL/6 strain, and dilatory responses to ANG II in C57BL/6 and to Phe in BALB/c. In the BALB/c strain, the presence of T and B cells promoted vasodilatory responses to Ado, attenuated dilations to low ACh concentrations, and exaggerated dilation and constriction responses to ANG II. Thus, under agonist challenge, NK cells generally promote constriction, whereas influences of T and B cells depend upon the stimulus. Therefore, lymphocytes or their products have physiological influences on microvascular arteriolar reactivity.

Cell-Based Assays Using Leukocyte Populations Isolated Directly From Whole Blood

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4842-4842
Author(s):  
Burgund Kauling ◽  
Volker Huppert ◽  
Stephanie Soltenborn ◽  
Angela Hillenkötter ◽  
Mariette Mohaupt ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
B Cells ◽  
Nk Cells ◽  
Whole Blood ◽  
Cell Isolation ◽  
Target Cells ◽  
A Cell ◽  
Leukocyte Populations ◽  
Gradient Separation

Abstract Abstract 4842 Obtaining pure and unaffected leukocyte populations is of utmost importance in diagnostic as well as research settings. So far, the isolation of functional leukocyte subpopulations from whole blood has been a time-consuming procedure, rendering the performance of downstream assays and analyses a challenging objective. We have developed a cell isolation technology that allows the purification of immune cells from human whole blood within 20 minutes. This novel technology requires a minimum of laboratory equipment. A cell isolation reagent is added to the anticoagulated blood sample and mixed briefly. While placed in a strong magnetic field, magnetically labeled non-target cells are depleted, while untouched target cells remain in the supernatant. Simultaneously, a reagent-assisted erythrocyte sedimentation phase occurs, which depletes ∼99.7 % of erythrocytes. Using this novel technology, Natural Killer cells, B cells, T cells, CD4+ T helper cells, CD8+ cytotoxic T cells and naïve B cells were isolated from 30mL of anticoagulated human whole blood. Target cells were recovered in a volume of 25–30 mL of supernatant (67% plasma, 33% Phosphate buffered saline) and average purities among white blood cells were 88.9% for NK cells, 88.2% for B cells, 97.8% for T cells, 93.0% for CD4+ cells, 78.9% for CD8+ T cells and 79.4% for naïve B cells, yields were 75.5%, 84.4%, 54.5%, 63.0%, 59.5% and 96.8% respectively (n >6 each). Red Blood cells were reduced by ∼99.7%, platelets by >99.9%. Cytotoxicity and proliferative capacity of isolated NK cells were measured in cytotoxicity assays with K562 target cells and proliferation assays with antibody loaded large magnetic beads respectively. Cytotoxicity and proliferation rate were comparable to those assessed using NK cells isolated by Ficoll density gradient separation or magnetic cell sorting (NK cell isolation kit). In vitro proliferation assays with total T cells, CD4+ T cells, CD8+ T cells or B cells revealed that proliferation rate was identical to that of target cells which were isolated by Ficoll density gradient separation and magnetic cell sorting. We furthermore compared the mRNA yields from cells isolated with either method (new technology vs. Miltenyi's isolation kits). The mRNA samples were subsequently subjected to gene expression analysis. Comparing the results obtained from samples isolated with the two different separation methods, we could not detect any significant differences in gene expression levels. These results demonstrate, that cells isolated with the novel whole blood cell isolation strategy, can be used for cell-based functional assays, as well as gene expression profiling. Additionally, overall processing time can be significantly reduced, which is highly desirable for sensitive downstream experiments. Disclosures: Kauling: Miltenyi Biotec GmbH: Employment. Huppert:Miltenyi Biotec GmbH: Employment. Soltenborn:Miltenyi Biotec GmbH: Employment. Hillenkötter:Miltenyi Biotec GmbH: Employment. Mohaupt:Miltenyi Biotec GmbH: Employment. Schmitz:Miltenyi Biotec GmbH: Employment.

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