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1.
Figure 2.

Figure 2. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Tryptophan and kynurenine concentration in cultures of IDO-expressing DCs. Native DCs (control), DCs infected with IDO-adeno-GFP, IDO-adeno, or native replication-defective adenoviruses (control) were cultured in RPMI 1640 plus 10% FCS. Tryptophan (A) and kynurenine (B) concentrations were determined and the values (μM/105 cells) are shown on the ordinate.

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
2.
Figure 4.

Figure 4. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Effect of kynurenine on T cell proliferation induced by allogeneic DCs or anti-CD3 antibody. Peripheral lymphocytes were stimulated with (A) allogeneic DCs or (B) anti-CD3 mAb for 6 and 3 d, respectively. Various amounts of kynurenine (abscissa) were added to the cultures. Positive control was performed without kynurenine. T cell proliferation was determined by 3[H]thymidine incorporation (cpm) (ordinate).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
3.
Figure 3.

Figure 3. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Reduction of allogeneic T cell stimulation by IDO-expressing DCs. Native DCs (pos. ctr.), DCs infected with IDO-adeno-GFP, IDO-adeno, or replication-defective adenoviruses (control) were coincubated with allogeneic peripheral blood lymphocytes for 6 d and cell proliferation measured by 3[H]thymidine incorporation (cpm) (ordinate). Negative controls consisted of DCs or lymphocytes only.

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
4.
Figure 8.

Figure 8. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Additive effect of tryptophan metabolites. Peripheral lymphocytes were stimulated with anti-CD3 antibody for 3 d and decreasing amounts of (A) kynurenine plus 3-hydroxykynurenine plus anthranilic acid plus 3-hydroxyanthranilic acid plus quinolinic acid, (B) kynurenine plus 3-hydroxykynurenine plus 3-hydroxyanthranilic acid, (C) kynurenine plus 3-hydroxykynurenine, (D) kynurenine plus 3-hydroxyanthranilic acid were added. All components of a mixture were added in equal amounts. The concentration of single substances is shown (abscissa).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
5.
Figure 7.

Figure 7. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Effect of IDO-induced tryptophan metabolites on T cell proliferation. Peripheral lymphocytes were stimulated with anti-CD3 antibody for 3 d in the presence of various amounts (abscissa) of (A) 3-hydroxykynurenine, (B) anthranilic acid, (C) 3-hydroxyanthranilic acid, or (D) quinolinic acid. Positive control consisted of T cell stimulation in the absence of metabolites. T cell proliferation was determined by 3[H]thymidine incorporation (cpm) (ordinate).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
6.
Figure 5.

Figure 5. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Polyclonal restimulation of kynurenine-suppressed T cells. Peripheral lymphocytes were stimulated with (A) anti-CD3 mAb in the presence of various amounts of kynurenine (abscissa). Positive control consisted of T cell stimulation in the absence of kynurenine. After 2 d the cells were washed and (B) restimulated with Con-A for another 3 d. T cell proliferation was determined by 3[H]thymidine incorporation (cpm) (ordinate).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
7.
Figure 11.

Figure 11. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Sensitivity of T, B, NK cells and DCs to tryptophan metabolites. T, B, and NK cells were separated from PBMCs and DCs were generated as described previously. The cells were incubated with a metabolite mixture (kynurenine plus 3-hydroxykynurenine plus anthranilic acid plus 3-hydroxyanthranilic acid plus quinolinic acid) (concentrations: 0, 8, 16, 32 μM for each metabolite). The percentage of dead cells (ordinate) was measured by 7-AAD-staining every day (abscissa).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
8.
Figure 6.

Figure 6. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Allogeneic restimulation of T cells activated with allogeneic DCs. Peripheral blood lymphocytes were stimulated with allogeneic DCs in the presence of decreasing amounts of kynurenine (abscissa). After 5 d the cells were extensively washed and restimulated with DCs from the same or unrelated donors. T cell proliferation was measured by 3[H]thymidine incorporation (cpm) (ordinate) after primary and secondary stimulation. The curves show the degree of proliferation after the first stimulation (top curve) and upon restimulation with third-party (intermediate) or donor-specific DCs (bottom curve).

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
9.
Figure 10.

Figure 10. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Time dependency of cytotoxicity and preferential killing of activated T cells. T cells were separated from PBMCs using magnetic beads and kept in culture (A) with anti-CD3 antibody or (B) without anti-CD3 antibody, in the presence of a mixture of tryptophan metabolites (kynurenine plus 3-hydroxykynurenine plus anthranilic acid plus 3-hydroxyanthranilic acid plus quinolinic acid) (concentrations: 0, 8, 16, 32 μM for each compound). The percentage of dead cells (ordinate) was measured every day (abscissa) by 7-AAD staining.

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
10.
Figure 1.

Figure 1. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Transcription of IDO-gene in eukaryotic cells infected with recombinant IDO-adenoviruses. mRNA was extracted from human embryonic retinoblast cells infected with IDO-adeno-GFP, IDO-adeno, or with replication-defective adenoviruses (neg. ctr.-a) and reverse transcribed into cDNA. PCR with IDO-specific primers was performed and the products analyzed by agarose gel electrophoresis. Positive control consisted of material extracted from recombinant IDO-adenoviruses and neg. ctr.-b of water instead of template. Lane 1: DNA molecular weight marker; lane 2: pos. ctr., 3, neg. ctr.-b; lane: 4, IDO-adeno-GFP; lane 5: IDO-adeno; lane 6: neg. ctr.-a; lane 7: DNA molecular weight marker. Lanes 4 and 5 show the relevant IDO-specific bands.

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.
11.
Figure 9.

Figure 9. From: Inhibition of Allogeneic T Cell Proliferation by Indoleamine 2,3-Dioxygenase-expressing Dendritic Cells.

Induction of cell death by tryptophan metabolites. Lymphocytes were stimulated with anti-CD3 in the presence or absence (control) of active metabolites (1,501 μM kynurenine, 349 μM 3-hydroxykynurenine, 510 μM 3-hydroxyanthranilic acid) or a metabolite mixture (kynurenine plus 3-hydroxykynurenine plus anthranilic acid plus 3-hydroxyanthranilic acid plus quinolinic acid)(64 μM of each compound). After 3 d the cells were washed, stained with anti-CD3-FITC antibody and 7-AAD. The percentages of dead (7-AAD-positive) T cells was determined in FACScan™. A shows the lymphocyte gate. B–F show the percentage of dead cells in the negative control and after treatment with kynurenine, 3-hydroxyanthranilic acid, 3-hydroxykynurenine, or metabolite mixture.

Peter Terness, et al. J Exp Med. 2002 August 19;196(4):447-457.

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