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2 Is a Critical Signaling Mediator for Murine NK Cell Activating Receptors1
* Department of Pathology and Immunology, and Department of Medicine and Howard Hughes Medical Institute, Washington University School of Medicine, St Louis, MO 63110
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Abstract |
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(PLC) is a key regulator of intracellular Ca2+ mobilization. Two isoforms of PLC have been identified, PLC1 and PLC2. Previously, in vitro studies indicated that activating NK cell receptors signal through both isoforms. However, PLC2 deficiency alone was sufficient to induce a substantial impairment of NK cell-mediated cytotoxicity in vitro. Why PLC2 is more important than PLC1 for NK cell activation and whether PLC2 is also critical for NK cell development, secretion of IFN-, and clearance of viral infections in vivo is not known. In this study, we report that PLC2 is the predominant isoform expressed in murine NK cells. PLC2 deficiency did not affect NK cell numbers in bone marrow and spleen, but acquisition of Ly49 receptors by NK cells was partially impaired. PLC2-deficient NK cells exhibited a dramatic impairment of cytolytic function and IFN- production upon ligation of activating receptors, whereas they did secrete IFN- in response to cytokines. Consequently, mice lacking PLC2 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 PLC2 is crucial for development of the NK cell receptor repertoire and signaling of activating NK cell receptors, mediating optimal NK cell function in vivo. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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-chain of FcRs (FcR), which mediates signaling for the low-affinity receptor for IgG (FcRIIIA), among others. Another ITAM-containing adapter protein, death-associated protein 12 (DAP12)3/killer cell-activating receptor-associated proteins, associates with a number of activating receptors, including Ly49D and Ly49H, which recognize the Chinese hamster MHC class I molecule Hm1-C4 (5) and the murine CMV (MCMV)3-encoded class I-like molecule m157 (6, 7), respectively. Engagement of these receptors results in tyrosine phosphorylation of the associated adaptors’ ITAMs, which recruit the protein tyrosine kinase Syk (1, 2, 3, 4, 8). This kinase activates multiple downstream signaling mediators, including linker for activation of T cells, SH2 domain-containing leukocyte protein of 76 kDa, c-Abl-SH3 domain-binding protein-2, phospholipase C- (PLC), PI3K, Vav family guanine nucleotide exchange factors, the low-m.w. GTP-binding protein Rho-Rac, and the Erk kinases. Collectively, these signaling mediators trigger gene transcription and the cellular programs for exocytosis of lytic granules that allow NK cells to lyse target cells and produce proinflammatory chemokines and cytokines, particularly IFN-.
Another NK cell activating receptor, NKG2D, mediates "natural cytotoxicity" against tumor cells that, like YAC-1, express the NKG2D ligands Rae1 and murine UL16-binding protein-like transcript 1 (9, 10). NKG2D associates with a unique transmembrane adapter protein, DAP10 (11), which lacks cytoplasmic ITAMs but contains a YINM motif that binds the p85 regulatory subunit of PI3K (11, 12) and Grb2 (12). DAP10 couples engagement of NKG2D with a Syk-independent pathway that involves a more restricted array of downstream mediators than do the ITAM-mediated pathways (13). However, these mediators—PI3K, SH2 domain-containing leukocyte protein of 76 kDa, Vav1, Rho family GTPases, and PLC—are sufficient for triggering release of lytic granules and inducing some IFN- production. In mouse, NKG2D exists as two isoforms with distinct cytoplasmic domains that mediate differential association with adaptors: the long isoform (NKG2D-L), associates only with DAP10, whereas the short isoform (NKG2D-S) associates with both DAP10 and DAP12 (14, 15). The relative predominance of NKG2D-L/DAP10 and NKG2D-S/DAP10-DAP12 signaling varies with NK cell activation (14, 15). NKG2D-L/DAP10 is particularly evident in NK cells isolated ex vivo and after prolonged culture with IL-2, whereas NKG2D-S/DAP10-DAP12 prevails early after IL-2 activation.
A common signaling mechanism shared by the ITAM and DAP10-mediated pathways is modulation of intracellular calcium ion concentration (Ca2+), which is necessary for reorientation of microtubules and the actin cytoskeleton, exocytosis of lytic granules, and transcriptional activation of cytokine genes. Intracellular Ca2+ is primarily regulated by PLC, which hydrolyzes membrane phosphatidylinositols into diacylglycerol and inositol 1,4,5-trisphosphate (16, 17, 18). Inositol 1,4,5-trisphosphate triggers the mobilization of Ca2+ from internal stores, resulting in a transient intracellular Ca2+ flux. This is followed by sustained Ca2+ influx from the extracellular medium through Ca2+-permeable channels in the plasma membrane. Two isoforms of PLC, PLC1 and PLC2, have been identified. PLC1 is ubiquitously expressed, whereas PLC2 is predominantly expressed in hemopoietic cells (19). Activation of PLC1/2 is mediated by PI3K, which phosphorylates the inositol ring of phosphatidylinositides in D3, generating, among others, phosphatidylinositol 3,4,5-trisphosphate (20, 21, 22). Phosphatidylinositol 3,4,5-trisphosphate binds the pleckstrin homology domain of PLC and other proteins, including the Tec family kinases, promoting their recruitment to the cell membrane and consequent activation. Tec kinases, as well as Src and Syk kinases, further contribute to PLC activation and Ca2+ mobilization by phosphorylating PLC (20, 21, 22).
Early biochemical studies in human NK cells demonstrated that engagement of FcRIIIa results in tyrosine phosphorylation and activation of both PLC1 and PLC2 (23, 24, 25, 26). Later studies in the rat NK cell line RNK-16 showed that Ly49D activates PLC1 and induces Ca2+ mobilization (8). Moreover, cross-linking of NKG2D/DAP10 in human NK cells triggers phosphorylation of PLC2, whereas pharmacological blockade of PLC inhibits NKG2D-mediated killing (13). Cumulatively, these in vitro studies suggest that NK cell activating receptors use both PLC1 and PLC2. However, analysis of PLC2-deficient mice revealed substantial impairment of FcRIIIa-mediated cytotoxicity as well as a significant reduction in "natural cytotoxicity" against the classical NK cell target YAC-1, despite the presence of intact PLC1 (27). This suggests that, at least in mice, PLC2 is more important than PLC1 for NK cell activation. Alternatively, PLC2 may have a major role in some signaling pathways, whereas others are regulated by PLC1. A precedent for the differential requirement of related isoforms in NK cell signaling was recently reported for Vav (28). Moreover, although previous reports indicate that PLC is required for optimal NK cell-mediated cytotoxicity in vitro, it is unclear whether PLC is necessary for NK cell development or other NK cell functions, particularly secretion of IFN-. Finally, whether PLC is required for NK cell function in vivo is not known. To address these questions, we investigated NK cell development in PLC2-deficient mice as well as signaling via the ITAM- and DAP10-mediated pathways in PLC2-deficient NK cells both in vitro and in vivo. Our results demonstrate that PLC2 is the predominant PLC isoform in murine NK cells. PLC2 is not required for the development of normal NK cell numbers in bone marrow and spleen, but does play a role in the acquisition of NK cell receptor repertoire, particularly the Ly49 receptors. PLC2 is essential for both ITAM- and DAP10-mediated pathways that trigger cytotoxicity and IFN- secretion. In contrast, PLC2 is not necessary for NK cell secretion of IFN- in response to cytokines. Hence, mice lacking PLC2 control MCMV infection less effectively than do wild-type (WT) animals, particularly in the spleen, where viral clearance predominantly depends on NK cell-mediated cytotoxicity.
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Materials and Methods |
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PLC2–/– mice (27) on a C57BL/6 background were kindly provided by J. Ihle (St. Jude Children’s Research Hospital, Memphis, TN). C57BL/6 mice were purchased from Taconic Farms.
Antibodies
Fluorochrome-conjugated mAbs included the following: allophycocyanin-anti-NK1.1 (PK136); PerCP-Cy5.5-anti-CD3 (145-2C11); PerCP-Cy5.5-anti-CD19 (1D3); PE-Cy5-anti-TCR
(H57-597); PE-anti-IFN- (XMG1.2); PE-anti-Ly49E/C (4D12); PE-anti-Ly49F (HBF-719); FITC-anti-Ly49I (YLI-90); and FITC-anti-NKG2A/C/E (20d5). All these Abs were from BD Pharmingen PE-anti-NK62D (Cx5) was purchased from ebioscience. PE-anti-Ly49H (3D10) and FITC-anti-Ly49A (JR9) and FITC-anti-Ly49D (4E4) have been previously described (7, 29). We also used PE-anti-CD43 (S7); FITC-anti-Mac-1 (m1/70); PE-anti-IL-2/IL-15R (CD122, TM-1); PE-anti-2B4 (CD244); and PE-anti-c-Kit (2B8) from BD Pharmingen (data not shown).
Cell preparation and flow cytometry
Single-cell suspensions were prepared from bone marrow and spleen. For flow cytometry, cells were first incubated with mAb 2.4G2 (American Type Culture Collection) to block nonspecific Ab binding and then stained with combinations of the indicated fluorochrome-conjugated mAbs. Stained cells were analyzed with a FACSCalibur (BD Biosciences). In some experiments, dead cells were excluded by staining with propidium iodide.
Biochemical analysis
WT NK cells were cultured 7 days in IL-2 and used for Western blot analysis. The cells were lysed in lysis buffer (1% Triton X-100, 50 mM Tris-HCl (pH 8), 150 mM NaCl, 5 mM EGTA, and 1.5 mM MgCl2) and protease inhibitors (5 mM NaF, 1 mM sodium orthovanadate, 10 µg/ml aprotinin, 4 µg/ml leupeptin, and 1 mM PMSF). After 15 min on ice, lysates were centrifuged for 15 min at 14,000 rpm, and supernatants were analyzed by SDS-PAGE, followed by immunoblotting with Abs specific for PLC1 (sc-81) and PLC2 (sc-407) (Santa Cruz Biotechnology).
Cytotoxicity assays
Murine NK cells were purified from spleens with DX5 microbeads (Miltenyi Biotec), yielding >90% NK1.1+CD3– cells. Purified NK cells were cultured for different intervals in IL-2 and tested against target cells (YAC-1, RMA-S, RMA-S-Rae1, CHO, Baf/3, Baf/3-Hm1-C4, Baf/3-m157, EL4) by standard 4-h chromium release assay as previously described (28). Cytotoxicity was blocked with anti-Ly49D (4E5; BD Pharmingen) or anti-NKG2D Abs. To measure Ab-dependent cellular cytotoxicity, EL-4 cells were incubated with anti-Thy1.2 mAb (30H12) before cytotoxicity.
Induction and detection of IFN-
To induce IFN- in vitro, spleen cells (107) from untreated mice were incubated in six-well plates coated with mAb against NK1.1, Ly49D, NKG2D (2–5 µg/ml) (A10) (29) for 1 h, and then further incubated in the presence of brefeldin A (Sigma-Aldrich) for an additional 5–8 h. Alternatively, spleen cells were incubated with YAC-1 cells (10:1 ratio) or IL-12 (1 ng/ml) plus IL-18 (10 ng/ml). Cells were then stained with allophycocyanin-anti-NK1.1 and FITC-anti-Ly49A to gate NK cells. In addition, cells were stained with PE-Cy5-anti-CD3, PE-Cy5-anti-CD19, and PE-Cy5-anti-TCR to exclude T and B cells. Stained cells were fixed with 2% paraformaldehyde and permeabilized in saponin buffer (5% FCS, 0.5% saponin, and 10 mM HEPES). Intracellular IFN- was detected with PE-conjugated anti-IFN- mAb.
Mouse infections and plaque assays
C57BL/6 and PLC2–/– mice were housed in a biosafety level 2 facility at Washington University in accordance with all federal and university policies. MCMV (ATCC VR-1399) was grown and titered on NIH 3T12 fibroblasts (ATCC CCL 164). Salivary gland-passaged stocks (sgMCMV) were prepared by infecting BALB/c mice with 105 PFU of tissue culture-passaged MCMV, and salivary glands were harvested and homogenized in DMEM (Invitrogen Life Technologies) with 10% BCS (HyClone). Mice were infected with 5 x 105 PFU of sgMCMV i.p., and organs were harvested 3 days postinfection and frozen in 1 ml of DMEM at –80°C. Titers were determined by plaque assay on 3T12 fibroblasts. After mechanical disruption by bead-beating (BioSpec), serial 10-fold dilutions were made and plated in triplicate on 3T12 monolayers in six-well plates (Costar). After infection in minimal medium at 37°C for 1 h, monolayers were overlaid with 3 ml of 5% DMEM-0.5% Noble agar and incubated for 6 days (37°C, 5% CO2) with one overlay of 2 ml at 3 days. At 6 days, plates were overlaid with 2.5% neutral red in DMEM-0.5% Noble agar. Plaques were counted 12 h later. Statistical significance of differences in viral titers between groups of mice was determined by unpaired t test using the Prism software.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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2
Human NK cells express both the PLC1 and PLC2 isoforms (24, 25, 26). However, expression of the two PLC isoforms in murine NK cells has not been determined. Therefore, we purified NK1.1+CD3– NK cells from splenocytes, cultured them in vitro for 7 days with IL-2, and assessed PLC1 and PLC2 expression by immunoblot analysis of cell lysates. Murine NK cells expressed high levels of PLC2, whereas PLC1 was barely detectable (Fig. 1). PLC1 was present in other murine cells, including the pro-B cell Baf/3 and the mastocytoma cell P815, as well as in human NK cells and T cells (Fig. 1). We conclude that murine NK cells predominantly express PLC2.
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2–/– NK cells
PLC2 is critical for normal development of B cells but is not required for T cell development (27). Whether PLC2 deficiency affects NK cell development is not known. NK cells develop in the bone marrow through sequential stages that are distinguished by cell surface expression of various markers, including the IL-2 and IL-15 receptor common subunit (CD122), the cytokine receptor c-Kit, the integrin CD11b (Mac-1), the leukosialin CD43, and the receptors 2B4 (CD244), NKG2A-C-E, and Ly49s (30, 31, 32). Although NK cells were present in equivalent numbers in spleen and bone marrow of PLC2–/– and WT mice (data not shown), NK cells expressing inhibitory and activating Ly49 receptors were significantly reduced in PLC2–/– mice (Fig. 2). All of the other markers, including NKG2D, NKG2A/C/E, CD122, c-Kit, CD43, and CD11b, were expressed on similar proportions of NK cells and at comparable levels in PLC2–/– and WT spleen and bone marrow NK cells (Fig. 2 and data not shown). Moreover, PLC2–/– and WT NK cells exhibited normal levels of perforin and granzymes transcripts (data not shown). Thus, PLC2 appears to be required only for NK cell acquisition of a normal Ly49 receptor repertoire. However, a partial impairment of Ly49 repertoire does not block NK cell development, functional maturation, and acquisition of other NK cell activating and inhibitory receptors.
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2 impairs FcRIIIA-, DAP12-, and DAP10-mediated cytotoxicity
NK cells lacking PLC2 do not effectively lyse Ab-coated target cells, which indicates that PLC2 is required for FcRIIIA/FcR-mediated signaling (27). Consistent with this report, we found that PLC2–/– NK cells inefficiently lysed EL-4 lymphoma cells coated with an anti-Thy1.2 Ab (Fig. 3). To assess the role of PLC2 in DAP12-mediated killing, we focused on Ly49D and Ly49H, both of which exclusively signal through DAP12. Ly49D mediates lysis of cells expressing the Chinese hamster class I molecules Hm1-C4, whereas Ly49H recognizes cells expressing the MCMV class I-like molecule m157. The ability of PLC2–/– NK cells to lyse CHO cells expressing Hm1-C4, Baf/3 cells transfected with either Hm1-C4 or m157 was dramatically reduced in comparison to WT NK cells (Fig. 3). Thus, PLC2 is required for DAP12-mediated signaling. Moreover, because both FcR and DAP12 contain a cytoplasmic ITAM, we conclude that PLC2 is required for ITAM-mediated signaling.
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2 in DAP10-mediated killing, we studied the function of NKG2D, which is the only NK cell receptor known to signal through DAP10. In mouse, NKG2D signals through DAP10 and DAP12. However, the relative involvement of DAP10 and DAP12 varies during NK cell culture with IL-2 (14, 15). NKG2D preferentially signals through DAP10 in NK cells isolated ex vivo. Early after IL-2 activation, NKG2D mainly signals through DAP12. After prolonged culture with IL-2, DAP10 signaling again becomes predominant. Given this, we analyzed NKG2D-mediated cytotoxicity in PLC2–/– and WT NK cells ex vivo and at late time points during culture with IL-2. Target cells included YAC-1 cells as well as the class I-deficient RMA-S cells transfected with the NKG2D ligand Rae1 (RMA-S-Rae1). Lack of PLC2 severely reduced the ability of NK cells to lyse cells expressing NKG2D ligands, whether NK cells were isolated ex vivo or cultured for 14–20 days in IL-2 (Fig. 4). Thus, PLC2 is absolutely required for NKG2D/DAP10-mediated signaling. NKG2D-mediated lysis was also significantly reduced at day 7 of NK cell culture, confirming that DAP12 signaling requires PLC2 (data not shown).
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2–/– mice could be due to impairment of integrin signaling, resulting in a defect of NK cell-target cell adhesion. To test this possibility, we mixed WT and PLC2–/– NK cells with YAC-1. After 20-min incubation at 37°C we measured formation of conjugates by two-color flow cytometry. Under these conditions, the conjugation frequency of PLC2–/– NK cells with target cells was similar to that obtained with WT NK cells (data not shown). Thus, PLC2–/– NK cells do not have an obvious defect in adhering to target cells.
NK cell secretion of IFN- is partially dependent on PLC2
NK cells produce IFN- in response to a variety of stimuli, including engagement of activating receptors and exposure to proinflammatory cytokines secreted by macrophages and dendritic cells during viral infections (33). To examine the role of PLC2 in NK cell secretion of IFN-, we isolated splenocytes from PLC2–/– and WT mice and stimulated them with plate-bound Abs specific for the activating receptors NK1.1, Ly49D, and NKG2D, which signal through distinct associated adaptors. NK1.1 signals through FcR, as does FcRIIIA (34); Ly49D associates exclusively with DAP12 (1, 2), whereas NKG2D uses both DAP10 and DAP12 (14, 15). NK cells were also stimulated by conjugation with YAC-1 cells, which trigger NKG2D as well as-yet-undefined activating receptors. Assessment of IFN- in NK cells by intracellular staining revealed that PLC2–/– NK cells produced very little or no IFN- in comparison to WT NK cells after each stimulation tested (Fig. 5). In contrast, incubation of splenocytes with both IL-12 and IL-18 effectively induced comparable amounts of IFN- in PLC2–/– and WT NK cells (data not shown). We conclude that PLC2 is required for NK cell production of IFN- induced by activating receptors, whereas cytokine-induced IFN- production is independent of PLC2.
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2 is required for the early control of MCMV infection
To investigate the role of PLC2 in NK cell antiviral activity in vivo, we infected WT C57BL/6, PLC2–/– mice, and PLC2+/– littermates with MCMV and harvested spleens and livers 3 days postinfection. Viral titers in the spleens from PLC2–/– mice were 
100-fold higher than in those from WT and PLC2+/– mice, and this difference was highly significant (Fig. 6); viral titers in the liver were slightly higher in PLC2–/– than in WT and PLC2+/– mice, but this difference was not statistically significant (Fig. 6). Because spleen viral titers at this early time point of infection reflect the efficiency of NK cell lysis of MCMV-infected cells through the m157/Ly49H pathway (6, 7, 35, 36), PLC2 most likely plays a significant role in NK cell control of viral replication by transducing Ly49H-mediated signals. Differences in liver viral titers in PLC2–/– mice were not as impressive as those observed in the spleen, most likely because IFN- primarily controls MCMV infection in the liver (37, 38) and PLC2–/– NK cells can still secrete IFN- in response to IL-12 and IL-18.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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2 is the predominant PLC isoform in murine NK cells, as it is in B cells. Although pro-B cell differentiation is blocked and mature B cells are reduced in numbers (27) in PLC2–/– mice, PLC2 is not required for NK cell expansion from bone marrow progenitors, because PLC2-deficient mice harbored normal numbers of NK cells in spleen and bone marrow. Moreover, PLC2-deficient cells expressed many markers indicative of maturation, including CD122, CD11b, CD43, c-Kit, NKG2D, NKG2A/C/E, perforin, and granzymes. However, we observed reduced percentages of NK cells expressing Ly49 receptors. Because acquisition of Ly49 receptors represents a relatively late event in the sequential process that leads to development of NK cells (39, 40, 41), a PLC2-mediated signal may be important for promoting the transition to the final developmental stage, which involves complete acquisition of Ly49 receptors. NK cell receptors acquired at an earlier stage of development, such as NKG2D and NK1.1, could trigger this PLC2-mediated pathway upon engaging cognate ligands on stromal cells.
Although PLC2 has a limited role in NK cell development, NK cell effector functions largely require PLC2. Both ITAM- and DAP10-coupled activating receptors substantially depend on PLC2 activation as a common signaling mechanism to trigger NK cell-mediated cytotoxicity. Supporting this conclusion, we found that PLC2 deficiency causes a remarkable reduction in both Ab-dependent cytotoxicity and "natural cytotoxicity" in vitro. The impaired cytotoxicity is not due to a defect in NK cell adhesion to target cells. Importantly, the defect in NK cell lysis is functionally significant in vivo; PLC2-deficient mice control MCMV infection in the spleen 100-fold less efficiently than do WT mice. In the context of previous studies demonstrating involvement of both PLC1 and PLC2 in human and rat NK cell activation, the magnitude of the defect caused by lack of PLC2 in murine NK cells was unexpected and is most likely explained by our observation that murine NK cells, unlike human NK cells, express PLC2 almost exclusively, along with barely detectable amounts of PLC1. These results document another significant difference between the human and mouse NK cell signaling pathways, supplementing the previously reported difference in NKG2D signaling (14, 15). Although PLC2 is absolutely essential for NK cell-mediated cytotoxicity, it is possible that the minimal amounts of PLC1 partially compensate for the lack of PLC2, mediating a residual cytotoxicity observed in some experiments. Consistent with this, it has been reported that PLC1 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 PLC2-independent pathways for exocytosis of lytic granules, such as the MAPK-mediated pathways (43, 44).
Previous studies in human NK cells showed that FcRIIIA-mediated secretion of IFN- is significantly dependent on intracellular Ca2+ mobilization, whereas cytokine-induced secretion of IFN- is Ca2+ independent (45). Because PLC2 is required for intracellular Ca2+ signaling, we expected significantly reduced IFN- secretion by PLC2–/– NK cells in response to engagement of activating receptors. Indeed, we found that PLC2–/– NK cells did not produce IFN- in comparison with WT NK cells following ligation of activating cell surface receptors. In contrast, in the presence of IL-12 and IL-18, NK cells produced IFN- in a PLC2-independent fashion. The partial maintenance of IFN- secretion in PLC2–/– mice together with the predominant role of IFN- in controlling MCMV infection in the liver (37, 38) explain why liver viral titers are not significantly increased by PLC2 deficiency.
In summary, we have shown that PLC2 is a downstream mediator crucial for signaling of all activating NK cell receptors. This signaling pathway promotes acquisition of the Ly49 cell receptors repertoire, receptor-mediated cytotoxicity, and receptor-mediated IFN- secretion and, ultimately, is required for optimal NK cell surveillance of MCMV infection in vivo.
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Acknowledgments |
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2–/– mice. |
Disclosures |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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Footnotes |
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1 This work was supported by National Institutes of Health Grant 5R01AI056139-03. Animal work was performed in a mouse facility supported by NCRP Grant CO6RRO12466. 
2 Address correspondence and reprint requests to Dr. Marco Colonna, Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110. E-mail address: mcolonna{at}pathbox.wustl.edu
3 Abbreviations used in this paper: DAP, DNAX-activating protein; MCMV, murine CMV; PLC, phospholipase C; WT, wild type.
Received for publication January 12, 2005. Accepted for publication May 5, 2005.
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References |
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2-microglobulin-deficient mice. J. Immunol. 169: 126-136. isoforms in haematopoietic cells: why one, not the other?. Cell. Signal. 13: 691-701.[Medline]
RIIIA results in phospholipase C-1 tyrosine phosphorylation and p56lck activation. J. Exp. Med. 176: 1745-1750. receptor activation induces the tyrosine phosphorylation of both phospholipase C (PLC)-1 and PLC-2 in natural killer cells. J. Exp. Med. 176: 1751-1755.RIIIA on natural killer cells results in tyrosine phosphorylation of PLC-1 and PLC-2. J. Immunol. 150: 2668-2674.[Abstract]
receptor-initiated activation of natural killer cells. J. Biol. Chem. 270: 16415-16421.2 is essential in the functions of B cell and several Fc receptors. Immunity 13: 25-35.[Medline]
is essential for activation signal through NKR-P1 (CD161) in natural killer (NK) cells and NK1.1+ T cells. J. Exp. Med. 186: 1957-1963. production and antiviral defense: studies of natural killer and T cell responses in contrasting viral infections. J. Immunol. 156: 1138-1142.[Abstract]
1 (PLC-1) to T-cell antigen receptor-mediated signaling: reconstitution studies of a PLC-1-deficient Jurkat T-cell line. Mol. Cell. Biol. 20: 9149-9161.R(CD16) interaction with ligand induces Ca2+ mobilization and phosphoinositide turnover in human natural killer cells: role of Ca2+ in FcR(CD16)-induced transcription and expression of lymphokine genes. J. Exp. Med. 169: 549-567.This article has been cited by other articles:
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H. Sasanuma, A. Tatsuno, S. Hidano, K. Ohshima, Y. Matsuzaki, K. Hayashi, C. A. Lowell, D. Kitamura, and R. Goitsuka Dual function for the adaptor MIST in IFN-{gamma} production by NK and CD4+NKT cells regulated by the Src kinase Fgr Blood, May 1, 2006; 107(9): 3647 - 3655. [Abstract] [Full Text] [PDF]
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S. Chiesa, M. Mingueneau, N. Fuseri, B. Malissen, D. H. Raulet, M. Malissen, E. Vivier, and E. Tomasello Multiplicity and plasticity of natural killer cell signaling pathways Blood, March 15, 2006; 107(6): 2364 - 2372. [Abstract] [Full Text] [PDF]
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A. Caraux, N. Kim, S. E. Bell, S. Zompi, T. Ranson, S. Lesjean-Pottier, M. E. Garcia-Ojeda, M. Turner, and F. Colucci Phospholipase C-{gamma}2 is essential for NK cell cytotoxicity and innate immunity to malignant and virally infected cells Blood, February 1, 2006; 107(3): 994 - 1002. [Abstract] [Full Text] [PDF]
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Y. T. Bryceson, M. E. March, H.-G. Ljunggren, and E. O. Long Synergy among receptors on resting NK cells for the activation of natural cytotoxicity and cytokine secretion Blood, January 1, 2006; 107(1): 159 - 166. [Abstract] [Full Text] [PDF]
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