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via Autocrine IL-121
Hepatobiliary Service, Memorial Sloan-Kettering Cancer Center, New York, NY 10021
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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than do DC, or even NK cells. We determined that CpG, but not IL-2, induces NKDC to secrete IFN- via the autocrine effects of IL-12. In vivo, CpG dramatically increases the number of NKDC. Furthermore, NKDC induce greater Ag-specific T cell activation than do DC after adoptive transfer. Their unique ability to lyse tumor cells, present Ags, and secrete inflammatory cytokines suggests that NKDC may play a crucial role in linking innate and adaptive immunity. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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, which is secreted in response to IL-12 originating from macrophages or dendritic cells (DC)3 (2). IFN- plays a critical role in bridging innate and adaptive immunity by increasing Ag presentation through the MHC class I and II pathways (3, 4). Transgenic mice lacking IFN- or its receptor have impaired immune responses to Mycobacterium bovis, Listeria monocytogenes, and vaccinia virus (5, 6, 7).
DC also contribute to innate immunity. Upon stimulation, DC produce inflammatory cytokines, such as IL-12 (8, 9) and TNF-
(10, 11). Recently, DC have also been shown to secrete IFN- in response to IL-12 alone or in combination with IL-18 (12, 13, 14, 15). Additionally, DC contribute to innate immune responses by activating NK cells via IL-12 secretion and direct cellular interaction (16, 17, 18, 19, 20). DC are established already as central mediators of adaptive immunity because they specialize in Ag capture and presentation to T cells (21).
Although NK cells and DC are known to have dichotomous phenotypic and functional features, there are few data on immunologic cells that possess characteristics of both effectors. Josien et al. (22) reported that rat-splenic DC, most of which express NK cell receptor protein 1, are able to lyse Yac-1 targets. Herein, we demonstrate that NK1.1+CD11c+ cells in the lymphoid and nonlymphoid organs of normal mice are immunostimulatory and share phenotypic and functional properties of both NK cells and DC. It is likely that these NKDC play a significant role in immunity because they are able to lyse targets, activate naive T cells, and, upon stimulation, secrete high levels of IFN-.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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Adult male C57BL/6 (H-2Kb) and BALB/c (H-2Kd) mice were purchased from Taconic Farms. Rag2–/–OT-I (OT-I) and Rag2–/–OT-II (OT-II) OVA TCR-transgenic mice (H-2Kb) were also obtained from Taconic Farms. Mice were maintained in the pathogen-free animal housing facility at Memorial Sloan-Kettering Cancer Center. All procedures were approved by the Institutional Animal Care and Use Committee.
Cell isolation
Liver nonparenchymal cells were isolated as previously described, with minor modifications (23, 24). Briefly, animals were euthanized by CO2 inhalation before laparotomy, then the portal vein was cannulated in situ with a 26-gauge needle (BD Biosciences) and perfused with 2 ml of 1% (w/v) collagenase D (Sigma-Aldrich) in PBS. The livers were then morselized and incubated in collagenase for 15 min at 37°C before they were passed through a sterile 100-µm nylon cell strainer (BD Falcon). Hepatocytes were eliminated by discarding the pellets from a series of three low-speed centrifugations (30 x g). The resulting nonparenchymal cell suspension was then pelleted (300 x g for 7 min), red cells were lysed using a hypotonic solution, and the cells were washed in medium (RPMI 1640, 10% FCS, 2 mM L-glutamine, 0.1% 2-ME, 100 U/ml penicillin, and 100 U/ml streptomycin). Single-cell suspensions of spleen, mesenteric lymph nodes, and thymus were also prepared by morselizing the tissue, incubating in collagenase, and passing through a 70-µm nylon cell strainer (BD Falcon). Following red cell lysis and washing, the resulting suspensions were sorted into CD11c+ and CD11c– fractions using immunomagnetic beads (Miltenyi Biotec) after blocking FcIII/IIRs with the mAb 2.4G2 (Fc block; 1 µg/million cells; mAb Core Facility, Sloan-Kettering Institute). The cells were then stained with fluorescently conjugated Abs to B220, NK1.1, and CD11c for additional separation of subtypes using a MoFlo cell sorter (DakoCytomation). From the CD11c+ fraction, NKDC were defined as NK1.1+CD11c+ and DC as NK1.1–CD11c+B220– (thereby excluding B220+ plasmacytoid DC). After immunomagnetic bead presorting and additional purification by FACS, we were able to isolate 
1–2 x 104 NKDC/spleen. NK cells were defined as NK1.1+CD11c– cells from the bead-presorted CD11c– fraction. Care was taken to exclude highly autofluorescent cells during FACS sorting, and sorted cells were typically >97% pure for the desired set of surface markers.
Flow cytometry
Four-color flow cytometry was performed on a FACSCalibur flow cytometer (BD Biosciences). Voltages were determined using unstained cells, and single-stained positive controls for each fluorochrome were used to set compensation. All samples were incubated with Fc block before staining. We labeled 5 x 105 cells with 0.1 µg of FITC, PE, allophycocyanin, or biotin-conjugated Ab (all BD Pharmingen). Biotinylated Abs were stained secondarily with streptavidin-PerCP. Cells were stained for FcR (2.4G2), CD45R/B220 (RA3-6B2), NK1.1 (PK136), CD11c (HL-3), MHC class II (I-Ab) (AF6-120.1), CD80 (16-10A1), CD86 (GL-1), CD8 (Ly-2), CD11b (M1/70), Ly-6G (RB6-8C5), CD3
(145-2C11), Ly49C/I (Ly49C) (5E6), CD244 (2B4), CD69 (H1.2F3), and CD49b (DX5).
Microscopy
FACS-sorted splenic NKDC, DC, and NK cells were studied by light and scanning electron microscopy (EM). For light micrographs, cells were spun at 600 rpm onto glass slides and fixed in formalin, followed by 100% methanol. Cells were then Giemsa stained and dehydrated before photographing at x630 magnification using a Zeiss Axiophot 2 microscope (Carl Zeiss). For EM, the cells were placed on poly-L-lysine-coated Thermanox plastic coverslips (Nalge Nunc International) and then fixed in 2.5% glutaraldehyde/2% paraformaldehyde in 0.075 M cacodylate buffer for 1 h, rinsed in buffer, and serially dehydrated in ethanol (50, 75, 95, and then 100%), followed by propylene oxide. The samples were then freeze-dried in a Pearce-Edwards dryer and coated subsequently with Gold/Palladium in a Technics Hummer IV sputtering system. The samples were photographed using a JEOL JSM 35 Scanning Electron Microscope (JEOL USA).
Cytokine secretion
The in vitro cytokine production by FACS-sorted NK cells, NKDC, and DC was assessed by culturing 3 x 105 cells/ml in a 96-well U-bottom tissue culture plate in 75 µl of medium. Thirty-six hours later, supernatant was harvested from all wells, and IFN- content was measured using a cytometric bead array (BD Biosciences), according to the manufacturer’s directions. IL-12 production was assayed by ELISA (R&D Systems) per the manufacturer’s protocol. The TLR 9 agonist CpG ODN 1826 (10 µg/ml; Oligos Etc.), rat anti-mouse IL-12 (p40/p70) mAb or its isotype (rat IgG1, 
; 10 µg/ml), IL-2 (20 ng/ml), IL-4 (20 ng/ml), or IL-12 (20 ng/ml; all R&D Systems) were added to some wells.
T cell assays
MLR was performed by combining gamma-irradiated (3000 rad) stimulator cells with allogeneic T cells. FACS-sorted NK cells, NKDC, and DC from C57BL/6 mice were added in various numbers to 1 x 105 BALB/c T lymphocytes purified using Thy1.2 (CD90.2)-immunomagnetic beads (Miltenyi Biotec), according to the manufacturer’s protocol, in 96-well U-bottom plates (BD Falcon) in a total of 200 µl of medium. On day 3, 30 µl of supernatant were harvested from the top wells, containing the highest numbers of stimulators, and tested for the presence of IL-2, IL-4, IL-5 IFN-, and TNF- by cytometric bead array (BD Biosciences). The cultures were then pulsed with [3H]thymidine (1 µCi/well), and radioactive uptake was measured 20 h later. To assess the effects of CpG and IL-4 on T cell activation, NK cells, NKDC, and DC were incubated for 2 h in medium or medium plus CpG (10 µg/ml) and IL-4 (20 ng/ml), washed twice, then resuspended in medium and added to allogeneic T cells as above. Ag-specific CD8+ T cell activation was assayed with OT-I CD8+-transgenic T cells specific for OVA257–264 peptide (25). Ag-specific CD4+ T cell activation was assayed with OT-II-transgenic T cells specific for OVA323–339 peptide (26). OT-I and OT-II T cells were isolated from splenocytes using Thy 1.2-immunomagnetic beads (Miltenyi Biotec). Gamma-irradiated stimulators were plated at various concentrations with OT-I or OT-II T cells (3 x 104/well) and the appropriate OVA peptide (1 µg/ml; Peptide Synthesis Core, Sloan-Kettering Institute) in a 96-well U-bottom plate (BD Falcon) for 3 days. Cytokine production and [3H]thymidine incorporation were measured as above. None of the MLR, OT-I, or OT-II cultures contained detectable levels of IL-4 or IL-5. In vivo T cell activation was tested by loading NKDC or DC with OVA257–264 peptide (10 µg/ml) for 1 h at 37°C, washing twice in medium, and injecting 1 x 104 cells into each footpad of OT-I mice. Three days later, popliteal nodes were harvested, and single-cell suspensions were prepared. Nodal cells (3 x 103/well) were then cultured (in a total volume of 200 µl) with various numbers of OVA257–264 peptide-loaded, gamma-irradiated, splenic DC. Proliferation was measured as above using [3H]thymidine on day 3.
NK-specific lysis and presentation of tumor Ags to T cells
FACS-sorted, splenic NK cells, NKDC, and DC were cocultured with 1 x 103 [51Cr]sodium chromate (PerkinElmer Life and Analytical Sciences)-labeled Yac-1 cells (American Type Culture Collection) for 6 h. Spontaneous release (no effectors) and maximum release (2% Triton X-100; Sigma-Aldrich) were also assayed. Percent-specific lysis was calculated as (cpm experimental – cpm spontaneous release) x 100/(cpm maximum release/cpm spontaneous release). We also wished to assess the combined ability of NKDC to lyse tumor cells and subsequently present Ags captured from the tumor cells to T cells. To do this, we incubated Yac-1 cells with FITC-conjugated OVA for 1 h before washing and irradiating them (7000 rad). The Yac-1 cells were then FACS sorted for live (4',6'-diamidino-2-phenylindole–) FITC-positive (containing OVA) cells before overnight incubation (5 x 105 cells) with FACS-sorted NKDC (2.5 x 105 cells). To control for the possibility of spontaneous OVA release by the Yac-1 cells, we separated NKDC and Yac-1 cells in some wells with a Transwell insert containing 0.4-µm pores (Corning Glass). After overnight culture, NKDC were isolated using CD11c-immunomagnetic beads. Various numbers of NKDC were then incubated with OT-II T cells (3 x 104/well), without adding additional OVA protein or peptide. Twenty-four-hour [3H]thymidine incorporation was measured 3 days later.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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We and others (23, 27) have previously noted that many CD11c+ cells in the livers of C57BL/6 mice also express the NK marker NK1.1. To determine whether these NKDC are unique to the liver, we searched for their presence in lymphoid organs. Direct flow cytometry analysis of cells from the spleen, mesenteric lymph nodes, and thymus initially proved difficult because of the rarity (<0.2% of total cells) of NKDC. However, using immunomagnetic bead enrichment of CD11c+ cells before flow cytometry, we identified a distinct population of NKDC in each organ (Fig. 1). Overall, the percentage of CD11c+ cells that coexpressed NK1.1+ was high in the liver (23%) but low in the spleen (4%), nodes (7%), and thymus (2%). Conversely, a small percentage of NK1.1+ cells coexpressed CD11c in the liver (10%), spleen (3%), nodes (4%), and thymus (0.4%) (Table I).
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We focused on spleen NKDC because they are most abundant in that organ and because spleen NK cells and DC are already well characterized. First, we compared the phenotype of NKDC to that of NK cells and DC. NKDC had intermediate size and granularity as measured by forward and side light scatter, respectively (Fig. 2a). On the basis of Giemsa staining of FACS-sorted cells, NK cells resembled small lymphocytes, with a high nuclear-cytoplasmic ratio and dense hyperchromatic nuclei, whereas DC had abundant multivacuolated cytoplasm and dispersed chromatin (Fig. 2b). NKDC had an intermediate phenotype, with a moderate amount of cytoplasm that was vacuolated slightly, and open chromatin nuclei. Scanning EM revealed that as with DC, NKDC have cytoplasmic ruffling and projections (Fig. 2c).
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Rs (Fig. 3). They had negligible staining for lineage markers associated with lymphoid DC (CD8), granulocytes (Ly6-G), or T cells (CD3). A very small proportion of NKDC had a low-level expression of the myeloid marker CD11b, in contrast to myeloid DC and macrophages, which usually express high levels. Some NKDC expressed the inhibitory NK cell receptor Ly-49C, but the majority of NKDC expressed CD69, 2B4, and DX5, each of which is typically associated with NK cells. Overall, then, NKDC had a low expression of characteristic molecules associated with APC but possessed several identifiers of NK cells.
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Because splenic NKDC had cytoplasmic vacuoles, ruffling, and projections as with DC, we wanted to know whether they could function as APC. We first tested the ability of splenic NKDC to activate allogeneic T cells in a MLR. Despite their relatively low expression of typical DC markers, we found that freshly sorted, gamma-irradiated, splenic NKDC were capable of stimulating naive allogeneic T cells to proliferate (Fig. 4a). On a per cell basis, NKDC were approximately one-third as potent as splenic DC. NKDC induced supernatant levels of T cell IFN- and IL-2 that were proportional to the degree of T cell proliferation (Fig. 4a). As expected, NK cells did not stimulate allogeneic T cells to proliferate.
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production by OT-I T cells and IL-2 by OT-II T cells. Furthermore, we demonstrated that NKDC are able to capture and process protein Ags, because when OVA protein was used instead of peptide, they promoted both OT-I and OT-II T cell proliferation, although again at approximately one-third the potency of DC (data not shown). To ascertain whether NKDC activated immunity in vivo as they had in vitro, we adoptively transferred OVA257–264 peptide-loaded NKDC or DC directly into the footpads of naive OT-I mice. Strikingly, popliteal lymph nodes cells harvested 3 days following the adoptive transfer of 104 NKDC proliferated more robustly upon in vitro restimulation (3 x 103 cells/well) than did nodal cells from mice treated with DC (Fig. 4d). Because of their expression of several classical NK markers, we next determined whether, in addition to their ability to activate T cells, NKDC had NK cell function. Using a standard chromium release assay, we found that NKDC were able to lyse Yac-1 lymphoma cells. Surprisingly, though, they accomplished more lysis than did NK cells (Fig. 5a). The addition of exogenous IL-2 (20 ng/ml) enabled NK cells to achieve a similar magnitude of lysis as NKDC (data not shown). As expected, DC did not lyse Yac-1 targets. Because we found that NKDC could lyse independently tumor cells and activate T cells, we set out to determine whether they could combine these disparate functions and thereby potentially bridge innate and adaptive immunity. We incubated NKDC with OVA-loaded Yac-1 cells overnight, then isolated the NKDC and measured their ability to activate Ag-specific OT-II T cells. Indeed, we found that NKDC were able to process and present Ags captured from Yac-1 cells and thereby induce proliferation of OT-II T cells (Fig. 5b). We used a Transwell insert in some wells to exclude that NKDC merely acquired OVA by its spontaneous release from Yac-1 cells.
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via autocrine IL-12 effects
Given that NKDC had lytic capability, we sought to establish whether, as with NK cells, they could also produce large quantities of IFN- upon stimulation. Testing a variety of conditions, we found that NKDC cultured in IL-2 plus IL-4 produced far more IFN- than did DC or even NK cells (Fig. 6a). When cultured in IL-2 alone, NKDC made low levels (60 pg/ml) of IFN-, whereas DC and NK cells had undetectable levels. None of the three cell types produced measurable IFN- when cultured in medium alone or with the addition of IL-4 alone. Because CpG oligodeoxynucleotides are known to be potent activators of DC cytokine production via TLR 9 ligation, we tested their effects on NKDC IFN- production. We found that culture of NK cells, DC, or NKDC with CpG alone did not induce IFN- production. However, in the presence of IL-4, CpG was an even more potent stimulus for IFN- secretion by NKDC than was IL-2 (Fig. 6a).
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secretion by NKDC in response to CpG and IL-4. We hypothesized that it might rely on autocrine IL-12 production because TLR ligands and IL-4 are known to increase DC IL-12 production (28). Furthermore, IL-12 stimulates IFN- production by NK cells and is believed to do the same in DC, particularly in concert with IL-4 (13, 29, 30). Although we found that supernatant from NKDC cultured with CpG and IL-4 contained low amounts of IL-12 (15 pg/ml), the addition of a neutralizing mAb to IL-12 markedly reduced IFN- production (Fig. 6b). Conversely, culture in IL-12 alone triggered IFN- production by both NKDC and NK cells, and this effect was magnified in the presence of IL-4. However, the combination of IL-2 and IL-4 did not appear to work via IL-12 because blocking IL-12 had minimal effects on NKDC IFN- production (Fig. 6b). Notably, DC made minimal IFN- under any of the conditions tested.
To establish whether activation of NKDC with CpG and IL-4 altered their immunostimulatory function, we performed additional MLR. Preincubation of NKDC with CpG and IL-4 increased their ability to induce T cell proliferation and cytokine production (Fig. 6c). However, the mechanism appeared to be independent of NKDC production of IFN- because blockade of IL-12 in the MLR cultures did not influence alloproliferation (data not shown).
CpG expands NKDC in vivo
Because CpG had such dramatic in vitro effects on NKDC, we hypothesized that it would have in vivo effects as well. To test this, we treated mice with two different doses of CpG and then examined their spleens 2 or 6 days later. Two days following administration of 10 µg of CpG, the number of splenic NKDC increased 5-fold above baseline (Fig. 7a). A dose of 100 µg resulted in even greater NKDC expansion (>10 times baseline) at 2 days. The number of NKDC remained elevated on day 6 after treatment with either dose of CpG. In contrast, the increases in NK cells and DC were more modest, peaking at 2.5- and 5-fold, respectively (Fig. 7b).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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. NKDC can also contribute to the acquired immune response because we showed that freshly isolated NKDC activate naive T cells in vitro. Stimulated NKDC were even more proficient at inducing T cell proliferation. The capacity of NKDC to behave as professional APC was substantiated additionally by their ability to cross-present Ags in the OT-I T cell assay when whole OVA protein was used. The ability of NKDC to activate T cells in vivo was demonstrated by adoptive transfer, whereupon NKDC induced Ag-specific T cell activation. Notably, we also showed that NKDC can bridge innate and adaptive immunity by processing and presenting Ags acquired from tumor cells.
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. Now, it is thought that CD8+ T cells, 
T cells, NKT cells, B cells, macrophages, and DC also secrete IFN- (4). Although numerous reports have documented that NK cells and DC each produce IFN-, our results suggest that some of the findings from previous work may be more appropriately attributed to NKDC. Undoubtedly, NKDC have been included in many previous NK cell or DC isolates due to overlap in their expression of surface markers. We found that when we used NK cell (IL-2 or IL-12) or DC (CpG) stimuli in combination with IL-4, NKDC always produced more IFN- than did NK cells or DC. In fact, NK1.1–CD11c+ DC made only barely detectable amounts of IFN-. Others have shown that IL-4 synergizes with IL-12 in activating IFN- production by DC (13). In contrast, we found this relation to be true for NKDC and NK cells but not for DC (Fig. 6b). Although we confirmed that IL-4 combined with either IL-2 or IL-12 induces NK cells to secrete IFN-, the effects were more pronounced in NKDC. A combination of IL-4 and CpG was the most potent stimulus of IFN- secretion by NKDC.
Because IL-4 is known to be a major regulator of DC IL-12 production (28), we postulated that the mechanism of IFN- secretion by NKDC depended on autocrine effects of IL-12. Although NKDC made only minimal amounts of IL-12, a neutralizing Ab to IL-12 abrogated their secretion of IFN-. Our data are consistent with those of Hochrein et al. (14), who demonstrated that as little as 5 pg/ml IL-12 or a combination of IL-4, GM-CSF, CpG, and rat IFN- can activate murine CD4–CD8–CD11c+ cells to produce IFN-. In that report, the authors speculated, but did not prove, that because IL-4, GM-CSF, CpG, and IFN- together induced IL-12, DC secretion of IFN- might depend on IL-12. Importantly, they isolated DC from splenocytes depleted of T cells, granulocytes, macrophages, and erythroid cells but not NK cells. Therefore, it is likely that a substantial proportion of the cells within the CD4–CD8–CD11c+ fraction were in fact NKDC, which essentially lack CD8 and CD4 expression (Fig. 3 and data not shown). In contrast to our data and those of Hochrein et al. (14), Ohteki et al. (12) found that the CD8+ subset of CD11c+I-A+ DC was responsible for IFN- production after IL-12 stimulation. This group later showed that the combination of IL-12 with either IL-4 or IL-18 induced equivalent IFN- secretion by CD8+ and CD8– DC subsets (13). As pointed out by Hochrein et al. (14), their (and our) exclusive use of freshly isolated cells without culture or the adherence and careful avoidance of contamination with highly autofluorescent macrophages may explain the discrepant findings.
Our investigation of splenic NK1.1+CD11c+ cells was prompted by our recent observation that they are abundant in the liver of normal mice (23). Subsequently, we learned of the report of Homann et al. (31), who had previously isolated DX5+CD11c+ splenocytes from lymphocytic choriomeningitis virus (LCMV)-infected mice and found them to behave as regulatory cells. The investigators found that CD40L blockade prevented the development of autoimmune diabetes following LCMV infection of transgenic mice expressing LCMV proteins in pancreatic islet cells. The mechanism occurred via the generation of Ag-specific DX5+CD11c+ splenocytes because adoptive transfer of these cells conferred protection against diabetes. In contrast, we studied the function of NKDC from normal mice and found them to be immunogenic uniformly in vitro and in vivo. NKDC lysed tumor cells and activated Ag-specific T cells, as did DX5+CD11c+ cells from LCMV-infected mice. We did find that stimulation was necessary to induce NKDC to secrete IFN-. It is unclear why DX5+CD11c+ splenocytes acted as suppressor cells, although it may be peculiar to the model of LCMV infection and CD40 blockade because a LCMV variant has been shown recently to directly alter DC function (32). The expansion of DX5+CD11c+ cells in vivo by LCMV in the model used by Homann et al. (31) and of NK1.1+CD11c+ cells by CpG in our experiments suggests that NKDC may represent a common pathway of immune regulation in multiple disease settings, including viral and bacterial infection.
Although they comprise a large proportion of liver CD11c+ cells, the absolute number of NKDC in the liver, and even in lymphoid organs, is quite small. Therefore, obtaining sufficient numbers of NKDC for broad study is problematic. In our experience, the actual yield (i.e., 10,000–20,000 splenic NKDC/mouse) using immunomagnetic bead enrichment followed by stringent FACS is 10–20% of theoretical yield based on flow cytometry analysis. Hopefully, defining the lineage of NKDC will lead to a way of obtaining more cells, although we have already found that NKDC are not expanded in vivo by GM-CSF (data not shown). There are data suggesting that NK cells and DC derive from a common pathway. Marquez et al. (33) identified a bipotential precursor in the human thymus. Meanwhile, murine thymic T/NK progenitors have been found to have DC potential, which is lost at a similar stage of differentiation as is their ability to become NK cells (34). Additional evidence of a developmental relationship between NK cells and DC is suggested by treatment with human Flt-3 ligand, which expands both murine DC (35) and NK cells (36); however, using endogenous overexpression of murine Flt-3 ligand, we were unable to confirm differential NK expansion (37).
Thus, NKDC are multifunctional cells present in lymphoid and nonlymphoid organs. They have a unique ability to lyse tumor cells, capture and process Ags, and activate naive T cells. Upon stimulation, they can also produce high levels of IFN- via the mechanism of autocrine IL-12. Their in vivo expansion after CpG treatment and diverse functional capabilities suggest that they play an important physiologic role in immunity.
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Disclosures |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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Acknowledgments |
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Footnotes |
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1 This work was supported in part by grants CA009501 (to V.P.) and DK068346 and CA094503 (to R.D.). 
2 Address correspondence and reprint requests to Dr. Ronald P. DeMatteo, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021. E-mail address: dematter{at}mskcc.org
3 Abbreviations used in this paper: DC, dendritic cell; EM, electron microscopy; LCMV, lymphocytic choriomeningitis virus.
Received for publication May 22, 2004. Accepted for publication December 22, 2004.
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L. Cook, N. Miyahara, N. Jin, J. M. Wands, C. Taube, C. L. Roark, T. A. Potter, E. W. Gelfand, R. L. O'Brien, and W. K. Born Evidence That CD8+ Dendritic Cells Enable the Development of {gamma}{delta} T Cells That Modulate Airway Hyperresponsiveness J. Immunol., July 1, 2008; 181(1): 309 - 319. [Abstract] [Full Text] [PDF]
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E. Ullrich, M. Bonmort, G. Mignot, B. Jacobs, D. Bosisio, S. Sozzani, A. Jalil, F. Louache, E. Bulanova, F. Geissman, et al. Trans-Presentation of IL-15 Dictates IFN-Producing Killer Dendritic Cells Effector Functions J. Immunol., June 15, 2008; 180(12): 7887 - 7897. [Abstract] [Full Text] [PDF]
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K. Pilbeam, P. Basse, L. Brossay, N. Vujanovic, R. Gerstein, A. N. Vallejo, and L. Borghesi The Ontogeny and Fate of NK Cells Marked by Permanent DNA Rearrangements J. Immunol., February 1, 2008; 180(3): 1432 - 1441. [Abstract] [Full Text] [PDF]
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J. P. McAleer, D. J. Zammit, L. Lefrancois, R. J. Rossi, and A. T. Vella The Lipopolysaccharide Adjuvant Effect on T Cells Relies on Nonoverlapping Contributions from the MyD88 Pathway and CD11c+ Cells J. Immunol., November 15, 2007; 179(10): 6524 - 6535. [Abstract] [Full Text] [PDF]
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 I. Caminschi, F. Ahmet, K. Heger, J. Brady, S. L. Nutt, D. Vremec, S. Pietersz, M. H. Lahoud, L. Schofield, D. S. Hansen, et al. Putative IKDCs are functionally and developmentally similar to natural killer cells, but not to dendritic cells J. Exp. Med., October 29, 2007; 204(11): 2579 - 2590. [Abstract] [Full Text] [PDF]
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A. L. Coelho, M. A. Schaller, C. F. Benjamim, A. Z. Orlofsky, C. M. Hogaboam, and S. L. Kunkel The Chemokine CCL6 Promotes Innate Immunity via Immune Cell Activation and Recruitment J. Immunol., October 15, 2007; 179(8): 5474 - 5482. [Abstract] [Full Text] [PDF]
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U. I. Chaudhry, G. Plitas, B. M. Burt, T. P. Kingham, J. R. Raab, and R. P. DeMatteo NK Dendritic Cells Expanded in IL-15 Exhibit Antitumor Responses In Vivo J. Immunol., October 1, 2007; 179(7): 4654 - 4660. [Abstract] [Full Text] [PDF]
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L. Chen, E. Calomeni, J. Wen, K. Ozato, R. Shen, and J.-X. Gao Natural killer dendritic cells are an intermediate of developing dendritic cells J. Leukoc. Biol., June 1, 2007; 81(6): 1422 - 1433. [Abstract] [Full Text] [PDF]
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R. Copin, P. De Baetselier, Y. Carlier, J.-J. Letesson, and E. Muraille MyD88-Dependent Activation of B220-CD11b+LY-6C+ Dendritic Cells during Brucella melitensis Infection J. Immunol., April 15, 2007; 178(8): 5182 - 5191. [Abstract] [Full Text] [PDF]
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G. Plitas, U. I. Chaudhry, T. P. Kingham, J. R. Raab, and R. P. DeMatteo NK Dendritic Cells Are Innate Immune Responders to Listeria monocytogenes Infection J. Immunol., April 1, 2007; 178(7): 4411 - 4416. [Abstract] [Full Text] [PDF]
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 S.-R. Chang, K.-J. Wang, Y.-F. Lu, L.-J. Yang, W.-J. Chen, Y.-H. Lin, H.-H. Chang, and S.-L. Wang Characterization of Early Gamma Interferon (IFN-{gamma}) Expression during Murine Listeriosis: Identification of NK1.1+ CD11c+ Cells as the Primary IFN-{gamma}-Expressing Cells Infect. Immun., March 1, 2007; 75(3): 1167 - 1176. [Abstract] [Full Text] [PDF]
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H. Fujii, J. D. Trudeau, D. T. Teachey, J. D. Fish, S. A. Grupp, K. R. Schultz, and G. S. D. Reid In vivo control of acute lymphoblastic leukemia by immunostimulatory CpG oligonucleotides Blood, March 1, 2007; 109(5): 2008 - 2013. [Abstract] [Full Text] [PDF]
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E. Ullrich, M. Bonmort, G. Mignot, N. Chaput, J. Taieb, C. Menard, S. Viaud, T. Tursz, G. Kroemer, and L. Zitvogel Therapy-Induced Tumor Immunosurveillance Involves IFN-Producing Killer Dendritic Cells Cancer Res., February 1, 2007; 67(3): 851 - 853. [Abstract] [Full Text] [PDF]
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D. Vremec, M. O'Keeffe, H. Hochrein, M. Fuchsberger, I. Caminschi, M. Lahoud, and K. Shortman Production of interferons by dendritic cells, plasmacytoid cells, natural killer cells, and interferon-producing killer dendritic cells Blood, February 1, 2007; 109(3): 1165 - 1173. [Abstract] [Full Text] [PDF]
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R. Yadav, D. J. Zammit, L. Lefrancois, and A. T. Vella Effects of LPS-mediated bystander activation in the innate immune system J. Leukoc. Biol., December 1, 2006; 80(6): 1251 - 1261. [Abstract] [Full Text] [PDF]
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U. I. Chaudhry, T. P. Kingham, G. Plitas, S. C. Katz, J. R. Raab, and R. P. DeMatteo Combined Stimulation with Interleukin-18 and CpG Induces Murine Natural Killer Dendritic Cells to Produce IFN-{gamma} and Inhibit Tumor Growth Cancer Res., November 1, 2006; 66(21): 10497 - 10504. [Abstract] [Full Text] [PDF]
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T. Aranami, S. Miyake, and T. Yamamura Differential Expression of CD11c by Peripheral Blood NK Cells Reflects Temporal Activity of Multiple Sclerosis J. Immunol., October 15, 2006; 177(8): 5659 - 5667. [Abstract] [Full Text] [PDF]
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 S.-i. Fujii, K. Shimizu, H. Hemmi, M. Fukui, A. J. Bonito, G. Chen, R. W. Franck, M. Tsuji, and R. M. Steinman Glycolipid {alpha}-C-galactosylceramide is a distinct inducer of dendritic cell function during innate and adaptive immune responses of mice PNAS, July 25, 2006; 103(30): 11252 - 11257. [Abstract] [Full Text] [PDF]
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V. Holl, M. Peressin, S. Schmidt, T. Decoville, S. Zolla-Pazner, A.-M. Aubertin, and C. Moog Efficient inhibition of HIV-1 replication in human immature monocyte-derived dendritic cells by purified anti-HIV-1 IgG without induction of maturation Blood, June 1, 2006; 107(11): 4466 - 4474. [Abstract] [Full Text] [PDF]
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 U. I. Chaudhry, S. C. Katz, T. P. Kingham, V. G. Pillarisetty, J. R. Raab, A. B. Shah, and R. P. DeMatteo In vivo overexpression of Flt3 ligand expands and activates murine spleen natural killer dendritic cells FASEB J, May 1, 2006; 20(7): 982 - 984. [Abstract] [Full Text] [PDF]
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C. A. Shaw and M. N. Starnbach Stimulation of CD8+ T Cells following Diphtheria Toxin-Mediated Antigen Delivery into Dendritic Cells Infect. Immun., February 1, 2006; 74(2): 1001 - 1008. [Abstract] [Full Text] [PDF]
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