Pivotal role of phosphoinositide-3 kinase in regulation of cytotoxicity in natural killer cells

Jiang, Kun; Zhong, Bin; Gilvary, Danielle L.; Corliss, Brian C.; Hong-Geller, Elizabeth; Wei, Sheng; Djeu, Julie Y.

doi:10.1038/80859

Cited by 314 publications

(342 citation statements)

References 32 publications

(37 reference statements)

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“…Consistent with this, it has been reported that PLC␥1 is capable of mediating a significant signaling function in T cells even after a 90% reduction (42). Murine NK cell functions may also partially rely on PLC␥2-independent pathways for exocytosis of lytic granules, such as the MAPK-mediated pathways (43,44).…”

Section: Discussionsupporting

confidence: 56%

Phospholipase C-γ2 Is a Critical Signaling Mediator for Murine NK Cell Activating Receptors

Tassi¹,

Presti²,

Kim

et al. 2005

The Journal of Immunology

106

104

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Phospholipase C-γ (PLCγ) is a key regulator of intracellular Ca2+ mobilization. Two isoforms of PLCγ have been identified, PLCγ1 and PLCγ2. Previously, in vitro studies indicated that activating NK cell receptors signal through both isoforms. However, PLCγ2 deficiency alone was sufficient to induce a substantial impairment of NK cell-mediated cytotoxicity in vitro. Why PLCγ2 is more important than PLCγ1 for NK cell activation and whether PLCγ2 is also critical for NK cell development, secretion of IFN-γ, and clearance of viral infections in vivo is not known. In this study, we report that PLCγ2 is the predominant isoform expressed in murine NK cells. PLCγ2 deficiency did not affect NK cell numbers in bone marrow and spleen, but acquisition of Ly49 receptors by NK cells was partially impaired. PLCγ2-deficient NK cells exhibited a dramatic impairment of cytolytic function and IFN-γ production upon ligation of activating receptors, whereas they did secrete IFN-γ in response to cytokines. Consequently, mice lacking PLCγ2 controlled murine CMV infection substantially less effectively than did wild-type animals, and this defect was most evident in the spleen, where viral clearance mostly depends on NK cell lytic function. These results demonstrate that PLCγ2 is crucial for development of the NK cell receptor repertoire and signaling of activating NK cell receptors, mediating optimal NK cell function in vivo.

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Section: Discussionsupporting

confidence: 56%

Phospholipase C-γ2 Is a Critical Signaling Mediator for Murine NK Cell Activating Receptors

Tassi¹,

Presti²,

Kim

et al. 2005

The Journal of Immunology

106

104

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“…Additionally, our results show that unstimulated peripheral blood NK cells constitutively express CCL5 at mRNA and protein levels and sequester it in secretory vesicles distinct from those containing the cytotoxins. The MAPK (ERK, JNK, and p38) and PI3K pathways are involved in triggering the polarization and secretion of cytolytic granules in NK cells (7,8,24,44,45). In this study, we show that of these pathways, only the JNK MAPK plays a role in the regulation of steady-state CCL5 mRNA levels in NK cells.…”

Section: Discussionmentioning

confidence: 63%

JNK MAPK Pathway Regulates Constitutive Transcription of CCL5 by Human NK Cells through SP1

Kumar

Hosse

Toerne

et al. 2009

The Journal of Immunology

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N atural killer cells have features similar to CD8 ϩ T cells in that they recognize and lyse virally infected and neoplastic cells; express receptors such as CD28, CD43, and 2B4; and contain cytolytic granules (1-6). The activation of NK surface receptors triggers microtubule organization, granule polarization, and cytotoxicity signals through both ERK2 and JNK1 phosphorylation pathways (7,8).The chemokine CCL5 has been suggested to act as an important mediator of diverse inflammatory processes (9 -11). In CTL, the protein is associated with the development and expression of effector function (12)(13)(14). In CD8 ϩ T cells, CCL5 transcription occurs late (3-5 days) after the activation of naive cells and is coincident with the up-regulation of perforin, the granzymes, and granulysin (15)(16)(17). A complex of nuclear proteins recruited to the promoter by Krü ppel-like factor 13 (KLF13) 3 controls the transcription of CCL5 in CTL (18,19). KLF13 is translationally regulated and appears several days after T cell activation. The p38 and ERK1/2 MAPK pathways control KLF13 gene expression, whereas pre-mRNA processing factor 4 homolog B, a member of the MAPK family, regulates KLF13 phosphorylation in T cells and by extension CCL5 expression (20,21). In NK cells, the MAPKs and PI3K pathways are also involved in triggering the polarized release of cytolytic granules (7,8,(22)(23)(24); however, the role of these pathways in the control of CCL5 expression in NK cells has not been investigated.We report in this study that unlike in T cells, CCL5 mRNA is constitutively transcribed and translated in peripheral blood NK cells. The CCL5 protein is localized to secretory vesicles that are distinct from the cytolytic granules. The constitutive transcription of CCL5 mRNA in NK cells is mediated through binding of the SP1 transcription factor to a site just upstream from the TATA box of the CCL5 gene. The JNK/MAPK pathway in turn controls SP1 expression and, through this, constitutive CCL5 transcription. Materials and Methods Abs, inhibitors, and oligonucleotidesPhospho-specific Abs that recognize the activated forms of phospho-stressactivated protein kinase/JNK (Thr 183 /Tyr 185 ) and inactive forms of stressactivated protein kinase/JNK were purchased from Cell Signaling Technology (catalog 9251 and 9252). The Abs for NF-B, p50, p52, p65, SP1, and SP3 were purchased from Upstate Biotechnology (catalog 06-886, 06-413, 06-418, 07-645, and 07-107, respectively), and ␤ actin for proteinloading control and KLF13 was purchased from Abcam (catalog ab8227 and ab1127). Primary Abs were as follows: mouse IgG2b-anti-human CCL5 VL1 or VL2 hybridoma supernatant (25), mouse IgG1 anti-human granzyme B (Serotec; catalog MCA2120PE), mouse anti-human perforin (BD Biosciences; catalog 556434), goat anti-mouse IgG1 CD3 Pacific Blue (DakoCytomation; PB982), goat anti-mouse IgG1 CD56 allophycocyanin conjugated (BD Biosciences; catalog IM2474), and rabbit anti-human CD8 (Neomarkers; catalog RM-9116-R7 The costs of publication of this article were defray...

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“…2,3 Several structurally distinct receptors have been implicated in the activation of NK-cell cytolytic machinery: when cross-linked by the corresponding ligands on target cell, they trigger multiple and intersecting signaling pathways responsible for functional activation. 4 A vast array of activating NK receptors belonging to different families are coupled to the lipid modifying enzymes phosphatidylinositol3-kinase (PI3K) and phospholipase C␥ (PLC␥), which provide signals critically required for the activation of the cytolytic machinery [5][6][7][8] ; notably, both enzymes use the membrane phospholipid phosphatidylinositol-4,5-bisphosphate (PIP2) as common substrate. 9 Besides its role as signaling intermediate, PIP2 also acts as critical regulator of various cellular processes, including actin remodeling, membrane and vesicle trafficking, adhesion, and ion transport.…”

Section: Introductionmentioning

confidence: 99%

“…42,43 The well-characterized PI3K3Rac13PAK13MEK-ERK1/2 pathway critically controls the polarization of lytic granules. 5 The lack For personal use only. on April 7, 2019. by guest www.bloodjournal.org From of perturbation of 2B4-dependent PI3K activity that we observed in PI5KI␣-and PI5KI␥-silenced cells suggests functional redundancy of ␣-and ␥-dependent PIP2 pools in providing PI3K substrate; in this regard, recent data showed that the reduction of PIP2 levels in dominant negative RhoA-expressing cells does not affect BCRinduced Akt phosphorylation.…”

mentioning

confidence: 99%

PI5KI-dependent signals are critical regulators of the cytolytic secretory pathway

Micucci

Capuano

Marchetti

et al. 2008

Blood

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Although membrane phospholipid phosphatidylinositol-4,5bisphosphate (PIP2) plays a key role as signaling intermediate and coordinator of actin dynamics and vesicle trafficking, it remains completely unknown its involvement in the activation of cytolytic machinery. By live confocal imaging of primary human natural killer (NK) cells expressing the chimeric protein GFP-PH, we observed, during effector-target cell interaction, the consumption of a preexisting PIP2 pool, which is critically required for the activation of cytolytic machinery. We identified type I phosphatidylinositol-4-phosphate-5-kinase (PI5KI) ␣ and ␥ isoforms as the enzymes responsible for PIP2 synthesis in NK cells. By hRNA-driven gene silencing, we observed that both enzymes are required for the proper activation of NK cytotoxicity and for inositol-1,4,5-trisphosphate (IP3) generation on receptor stimulation. In an attempt to elucidate the specific step controlled by PI5KIs, we found that lytic granule secretion but not polarization resulted in impaired PI5KI␣-and PI5KI␥-silenced cells. Our findings delineate a novel mechanism implicating PI5KI␣ and PI5KI␥ isoforms in the synthesis of PIP2 pools critically required for IP3-dependent Ca 2؉ IntroductionNatural killer (NK) cells and cytotoxic T lymphocytes are critical effectors in the defense against tumor and viral infections 1 ; they exert cytotoxic function through the polarized secretion of granules containing proteolytic molecules, such as perforin and granzymes. This process involves several steps, including the formation of a cytolytic synapse between cytolytic effector and target cell, the rapid reorientation of the microtubule-organizing center along with lytic granules toward the target contact area followed by granule docking and fusion at specialized secretory domains within the cytolytic synapse. 2,3 Several structurally distinct receptors have been implicated in the activation of NK-cell cytolytic machinery: when cross-linked by the corresponding ligands on target cell, they trigger multiple and intersecting signaling pathways responsible for functional activation. 4 A vast array of activating NK receptors belonging to different families are coupled to the lipid modifying enzymes phosphatidylinositol3-kinase (PI3K) and phospholipase C␥ (PLC␥), which provide signals critically required for the activation of the cytolytic machinery [5][6][7][8] ; notably, both enzymes use the membrane phospholipid phosphatidylinositol-4,5-bisphosphate (PIP2) as common substrate. 9 Besides its role as signaling intermediate, PIP2 also acts as critical regulator of various cellular processes, including actin remodeling, membrane and vesicle trafficking, adhesion, and ion transport. 10,11 Surprisingly, the role of PIP2 and its regulatory mechanisms in lymphocyte-mediated cytotoxicity remain completely undefined.The main cellular source of PIP2 are type I phosphatidylinositol-4-phosphate-5-kinase (PI5KI) family members which phosphorylate PI4P on the D5 position of the inositol ring. Three major PI5KI is...

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Pivotal role of phosphoinositide-3 kinase in regulation of cytotoxicity in natural killer cells

Cited by 314 publications

References 32 publications

Phospholipase C-γ2 Is a Critical Signaling Mediator for Murine NK Cell Activating Receptors

Phospholipase C-γ2 Is a Critical Signaling Mediator for Murine NK Cell Activating Receptors

JNK MAPK Pathway Regulates Constitutive Transcription of CCL5 by Human NK Cells through SP1

PI5KI-dependent signals are critical regulators of the cytolytic secretory pathway

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