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Rat spleen dendritic cells express natural killer cell receptor protein 1 (NKR-P1) and have cytotoxic activity to select targets via a Ca2+-dependent mechanism.

Josien R, Heslan M, Soulillou JP, Cuturi MC - J. Exp. Med. (1997)

Bottom Line: We show that both spleen and thymus DC express the natural killer cell receptor protein 1 (NKR-P1) as a disulfide linked homodimer of 60 kD.Freshly isolated DC express a low level of NKR-P1, which is strongly upregulated after overnight culture.Spleen, but not thymus DC, were able to kill the NK-sensitive YAC-1 cell line in vitro, and since this killing was Ca2+ dependent, a Fas ligand-Fas interaction was probably not involved.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Sante et de la Recherche Medicale U437, Institut de Transplantation et de Recherche en Transplantation, Immeuble Jean Monnet, Nantes, France. josienr@rockvax.rockefeller.edu

ABSTRACT
Dendritic cells (DC) are a subset of leukocytes whose major function is antigen presentation. We investigated the phenotype and function of enriched (95-98.5%) rat DC. We show that both spleen and thymus DC express the natural killer cell receptor protein 1 (NKR-P1) as a disulfide linked homodimer of 60 kD. Freshly isolated DC express a low level of NKR-P1, which is strongly upregulated after overnight culture. Spleen, but not thymus DC, were able to kill the NK-sensitive YAC-1 cell line in vitro, and since this killing was Ca2+ dependent, a Fas ligand-Fas interaction was probably not involved. Besides their potent antigen-presenting function, DC can thus be cytotoxic for some tumor targets.

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Related in: MedlinePlus

In vitro cytolytic activity of spleen and thymus DC against  YAC-1 and P815. 2 × 103 51Cr-labeled YAC-1 (left) or P815 (right) target cells were incubated with total spleen cells (open circle), purified (95– 98.5) spleen (closed square), or thymus (open square) DC in triplicate for 6 h  at 37°C. 51Cr release was then assessed in the supernatant. Results of one  experiment representative of at least eight for each target are shown.
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Figure 4: In vitro cytolytic activity of spleen and thymus DC against YAC-1 and P815. 2 × 103 51Cr-labeled YAC-1 (left) or P815 (right) target cells were incubated with total spleen cells (open circle), purified (95– 98.5) spleen (closed square), or thymus (open square) DC in triplicate for 6 h at 37°C. 51Cr release was then assessed in the supernatant. Results of one experiment representative of at least eight for each target are shown.

Mentions: Based on our results that showed that rat DC expressed a NK-related molecule, we tested in 6-h Cr release assays the potential cytolytic activity of highly purified DC on target cells usually used for testing NK activity. As shown in Fig. 4, spleen DC efficiently killed the prototypical NK-sensitive YAC-1 cells. The level of killing observed was roughly similar to that reported with the rat NK cell line RNK-16 at the same E/T ratios (19). This unusual activity was also observed using spleen DC maintained in culture for >3 d (data not shown) and was reproduced in >10 independent experiments. The possibility that our DC preparations contained NK cells was excluded because the only non-DC contaminating our DC preparations (95–98.5% pure) corresponded to CD45− cells (fibroblasts, endothelial cells) and because our procedure included a depletion of CD8+ cells (20). Even if the contaminating cells were NK, it does not seem possible that they could be responsible for the lysis of YAC cells we observed as total spleen cells from the same animal, which contain 15–25% NK cells, exhibited an anti-YAC cytotoxic activity lower than that found for spleen DC at the same E/T ratios (Fig. 4). As compared to YAC-1 targets, the cytotoxic activity of spleen DC against P815 (Fig. 4), L12.10, or K562 (data not shown) was low.


Rat spleen dendritic cells express natural killer cell receptor protein 1 (NKR-P1) and have cytotoxic activity to select targets via a Ca2+-dependent mechanism.

Josien R, Heslan M, Soulillou JP, Cuturi MC - J. Exp. Med. (1997)

In vitro cytolytic activity of spleen and thymus DC against  YAC-1 and P815. 2 × 103 51Cr-labeled YAC-1 (left) or P815 (right) target cells were incubated with total spleen cells (open circle), purified (95– 98.5) spleen (closed square), or thymus (open square) DC in triplicate for 6 h  at 37°C. 51Cr release was then assessed in the supernatant. Results of one  experiment representative of at least eight for each target are shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2198993&req=5

Figure 4: In vitro cytolytic activity of spleen and thymus DC against YAC-1 and P815. 2 × 103 51Cr-labeled YAC-1 (left) or P815 (right) target cells were incubated with total spleen cells (open circle), purified (95– 98.5) spleen (closed square), or thymus (open square) DC in triplicate for 6 h at 37°C. 51Cr release was then assessed in the supernatant. Results of one experiment representative of at least eight for each target are shown.
Mentions: Based on our results that showed that rat DC expressed a NK-related molecule, we tested in 6-h Cr release assays the potential cytolytic activity of highly purified DC on target cells usually used for testing NK activity. As shown in Fig. 4, spleen DC efficiently killed the prototypical NK-sensitive YAC-1 cells. The level of killing observed was roughly similar to that reported with the rat NK cell line RNK-16 at the same E/T ratios (19). This unusual activity was also observed using spleen DC maintained in culture for >3 d (data not shown) and was reproduced in >10 independent experiments. The possibility that our DC preparations contained NK cells was excluded because the only non-DC contaminating our DC preparations (95–98.5% pure) corresponded to CD45− cells (fibroblasts, endothelial cells) and because our procedure included a depletion of CD8+ cells (20). Even if the contaminating cells were NK, it does not seem possible that they could be responsible for the lysis of YAC cells we observed as total spleen cells from the same animal, which contain 15–25% NK cells, exhibited an anti-YAC cytotoxic activity lower than that found for spleen DC at the same E/T ratios (Fig. 4). As compared to YAC-1 targets, the cytotoxic activity of spleen DC against P815 (Fig. 4), L12.10, or K562 (data not shown) was low.

Bottom Line: We show that both spleen and thymus DC express the natural killer cell receptor protein 1 (NKR-P1) as a disulfide linked homodimer of 60 kD.Freshly isolated DC express a low level of NKR-P1, which is strongly upregulated after overnight culture.Spleen, but not thymus DC, were able to kill the NK-sensitive YAC-1 cell line in vitro, and since this killing was Ca2+ dependent, a Fas ligand-Fas interaction was probably not involved.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Sante et de la Recherche Medicale U437, Institut de Transplantation et de Recherche en Transplantation, Immeuble Jean Monnet, Nantes, France. josienr@rockvax.rockefeller.edu

ABSTRACT
Dendritic cells (DC) are a subset of leukocytes whose major function is antigen presentation. We investigated the phenotype and function of enriched (95-98.5%) rat DC. We show that both spleen and thymus DC express the natural killer cell receptor protein 1 (NKR-P1) as a disulfide linked homodimer of 60 kD. Freshly isolated DC express a low level of NKR-P1, which is strongly upregulated after overnight culture. Spleen, but not thymus DC, were able to kill the NK-sensitive YAC-1 cell line in vitro, and since this killing was Ca2+ dependent, a Fas ligand-Fas interaction was probably not involved. Besides their potent antigen-presenting function, DC can thus be cytotoxic for some tumor targets.

Show MeSH
Related in: MedlinePlus