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Human dendritic cells mediate cellular apoptosis via tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

Fanger NA, Maliszewski CR, Schooley K, Griffith TS - J. Exp. Med. (1999)

Bottom Line: TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells.Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)alpha(+) blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c(+) DC and IL-3Ralpha(+) pre-DC subsets.These results indicate that TRAIL may serve as an innate effector molecule on CD11c(+) DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Discovery Research, Immunex Corporation, Seattle, Washington 98101, USA.

ABSTRACT
TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells. In this study, we demonstrate that human CD11c(+) blood dendritic cells (DCs) express TRAIL after stimulation with either interferon (IFN)-gamma or -alpha and acquire the ability to kill TRAIL-sensitive tumor cell targets but not TRAIL-resistant tumor cells or normal cell types. The DC-mediated apoptosis was TRAIL specific, as soluble TRAIL receptor blocked target cell death. Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)alpha(+) blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c(+) DC and IL-3Ralpha(+) pre-DC subsets. These results indicate that TRAIL may serve as an innate effector molecule on CD11c(+) DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

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Surface phenotype of freshly isolated CD11c+ and IL3Rα+ pre-DC subsets. (A) The two blood DCs were distinguishable based on CD11c and IL-3Rα expression as determined by multicolor flow cytometry. The CD11c+ and IL-3Rα+ pre-DCs were compared with Mφ for CD14, CD33, and HLA-DR expression. Filled histograms represent staining with specific antibody; open histograms represent isotype-matched controls. (B) CD11c+ DCs, but not IL-3Rα+ pre-DCs, express CD83 after 12-h incubation in complete medium. Histograms and contour plots represent 104 gated DCs, and viability was >95% as assessed by propidium iodide exclusion.
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Figure 1: Surface phenotype of freshly isolated CD11c+ and IL3Rα+ pre-DC subsets. (A) The two blood DCs were distinguishable based on CD11c and IL-3Rα expression as determined by multicolor flow cytometry. The CD11c+ and IL-3Rα+ pre-DCs were compared with Mφ for CD14, CD33, and HLA-DR expression. Filled histograms represent staining with specific antibody; open histograms represent isotype-matched controls. (B) CD11c+ DCs, but not IL-3Rα+ pre-DCs, express CD83 after 12-h incubation in complete medium. Histograms and contour plots represent 104 gated DCs, and viability was >95% as assessed by propidium iodide exclusion.

Mentions: The enriched “bulk” DCs comprised two distinct subsets distinguishable by the surface expression of CD11c and IL-3Rα (Fig. 1 A). The ratio of these two cell populations varied by donor, with some donors demonstrating as high as 70% CD11c+ DCs and 30% IL-3Rα+ pre-DCs, whereas others demonstrated the reverse percentages. Neither CD11c+ DCs nor IL-3Rα+ pre-DCs expressed CD14, but they did differentially express CD33 and HLA-DR. After 12 h of culture in complete medium, the levels of CD11c and CD33 remained relatively constant, whereas CD83 expression was detected on the majority (>90%) of CD11c+ DCs (Fig. 1 B). In contrast, the IL-3Rα+ pre-DCs expressed lower levels of CD33 and HLA-DR and failed to express CD83 after 12-h incubation under identical conditions (Fig. 1a and Fig. b). Distinct morphological differences were also observed between the two DC subsets. The CD11c+ DCs exhibited a characteristic multilobulated nucleus, in contrast to the IL-3Rα+ pre-DCs, which displayed an immature appearance consisting of an oval nucleus (Fig. 2A and Fig. B). Highly pure CD11c+ DCs and IL-3Rα+ pre-DCs were rapidly obtained using this method for functional analysis of TRAIL.


Human dendritic cells mediate cellular apoptosis via tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

Fanger NA, Maliszewski CR, Schooley K, Griffith TS - J. Exp. Med. (1999)

Surface phenotype of freshly isolated CD11c+ and IL3Rα+ pre-DC subsets. (A) The two blood DCs were distinguishable based on CD11c and IL-3Rα expression as determined by multicolor flow cytometry. The CD11c+ and IL-3Rα+ pre-DCs were compared with Mφ for CD14, CD33, and HLA-DR expression. Filled histograms represent staining with specific antibody; open histograms represent isotype-matched controls. (B) CD11c+ DCs, but not IL-3Rα+ pre-DCs, express CD83 after 12-h incubation in complete medium. Histograms and contour plots represent 104 gated DCs, and viability was >95% as assessed by propidium iodide exclusion.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195665&req=5

Figure 1: Surface phenotype of freshly isolated CD11c+ and IL3Rα+ pre-DC subsets. (A) The two blood DCs were distinguishable based on CD11c and IL-3Rα expression as determined by multicolor flow cytometry. The CD11c+ and IL-3Rα+ pre-DCs were compared with Mφ for CD14, CD33, and HLA-DR expression. Filled histograms represent staining with specific antibody; open histograms represent isotype-matched controls. (B) CD11c+ DCs, but not IL-3Rα+ pre-DCs, express CD83 after 12-h incubation in complete medium. Histograms and contour plots represent 104 gated DCs, and viability was >95% as assessed by propidium iodide exclusion.
Mentions: The enriched “bulk” DCs comprised two distinct subsets distinguishable by the surface expression of CD11c and IL-3Rα (Fig. 1 A). The ratio of these two cell populations varied by donor, with some donors demonstrating as high as 70% CD11c+ DCs and 30% IL-3Rα+ pre-DCs, whereas others demonstrated the reverse percentages. Neither CD11c+ DCs nor IL-3Rα+ pre-DCs expressed CD14, but they did differentially express CD33 and HLA-DR. After 12 h of culture in complete medium, the levels of CD11c and CD33 remained relatively constant, whereas CD83 expression was detected on the majority (>90%) of CD11c+ DCs (Fig. 1 B). In contrast, the IL-3Rα+ pre-DCs expressed lower levels of CD33 and HLA-DR and failed to express CD83 after 12-h incubation under identical conditions (Fig. 1a and Fig. b). Distinct morphological differences were also observed between the two DC subsets. The CD11c+ DCs exhibited a characteristic multilobulated nucleus, in contrast to the IL-3Rα+ pre-DCs, which displayed an immature appearance consisting of an oval nucleus (Fig. 2A and Fig. B). Highly pure CD11c+ DCs and IL-3Rα+ pre-DCs were rapidly obtained using this method for functional analysis of TRAIL.

Bottom Line: TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells.Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)alpha(+) blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c(+) DC and IL-3Ralpha(+) pre-DC subsets.These results indicate that TRAIL may serve as an innate effector molecule on CD11c(+) DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Discovery Research, Immunex Corporation, Seattle, Washington 98101, USA.

ABSTRACT
TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells. In this study, we demonstrate that human CD11c(+) blood dendritic cells (DCs) express TRAIL after stimulation with either interferon (IFN)-gamma or -alpha and acquire the ability to kill TRAIL-sensitive tumor cell targets but not TRAIL-resistant tumor cells or normal cell types. The DC-mediated apoptosis was TRAIL specific, as soluble TRAIL receptor blocked target cell death. Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)alpha(+) blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c(+) DC and IL-3Ralpha(+) pre-DC subsets. These results indicate that TRAIL may serve as an innate effector molecule on CD11c(+) DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

Show MeSH
Related in: MedlinePlus