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Activation of natural killer cells and dendritic cells upon recognition of a novel CD99-like ligand by paired immunoglobulin-like type 2 receptor.

Shiratori I, Ogasawara K, Saito T, Lanier LL, Arase H - J. Exp. Med. (2004)

Bottom Line: Transcripts of PILR ligand are present in many tissues, including some T cell lines.Cells expressing the PILR ligand specifically activated NK cells and dendritic cells that express the activating PILRbeta.Our findings reveal a new regulatory mechanism of innate immunity by PILR and its CD99-like ligand.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuoku, 260-8670, Japan.

ABSTRACT
Paired receptors that consist of highly related activating and inhibitory receptors are widely involved in the regulation of the immune system. Here, we report a mouse orthologue of the human activating paired immunoglobulin-like type 2 receptor (PILR) beta, which was cloned from a cDNA library of natural killer (NK) cells based on its ability to associate with the DAP12 signaling adaptor protein. The activating PILRbeta was expressed not only on NK cells but also on dendritic cells and macrophages. Furthermore, we have identified a novel CD99-like molecule as a ligand for the activating PILRbeta and inhibitory PILRalpha receptors. Transcripts of PILR ligand are present in many tissues, including some T cell lines. Cells expressing the PILR ligand specifically activated NK cells and dendritic cells that express the activating PILRbeta. Our findings reveal a new regulatory mechanism of innate immunity by PILR and its CD99-like ligand.

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Activation of DCs by PILR-L–expressing cells. 5 × 105 BM-derived DCs were cocultured with PILR-L–transfected 293T cells (closed circles) or parental 293T cells (open circles) at the indicated cell number for 1 d. BM-derived DCs were also stimulated with 1 μg/ml LPS as a positive control. Nitric oxide and mouse TNF-α produced in the culture supernatants were measured. Amounts of nitric oxide and TNF-α produced by DCs cultured in medium only were 1.5 ± 0.1 μM and 48.5 ± 7.7 pg/ml, respectively, and those produced by DCs upon LPS stimulation were 59.1 ± 2.8 μM and 2048 ± 127.9 pg/ml, respectively.
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fig8: Activation of DCs by PILR-L–expressing cells. 5 × 105 BM-derived DCs were cocultured with PILR-L–transfected 293T cells (closed circles) or parental 293T cells (open circles) at the indicated cell number for 1 d. BM-derived DCs were also stimulated with 1 μg/ml LPS as a positive control. Nitric oxide and mouse TNF-α produced in the culture supernatants were measured. Amounts of nitric oxide and TNF-α produced by DCs cultured in medium only were 1.5 ± 0.1 μM and 48.5 ± 7.7 pg/ml, respectively, and those produced by DCs upon LPS stimulation were 59.1 ± 2.8 μM and 2048 ± 127.9 pg/ml, respectively.

Mentions: BM-DCs were generated by culturing 5 × 106 BM cells with RPMI 1640 medium containing 10% FCS and 200 U/ml mouse GM-CSF for 7–8 d in six-well culture plates. Nonadherent cells were collected and cultured for 1 d. More than 95% of the cells expressed CD11b and CD11c. 5 × 105 BM-DCs were cocultured with various numbers of parental or PILR-L–transfected 293T cells in 48-well plates, as indicated in Fig. 8. After 24 h, culture supernatants were collected, and amounts of NO2 or TNFα were measured by using the Griess reagent (Sigma-Aldrich) or a TNFα ELISA kit (Genzyme), respectively.


Activation of natural killer cells and dendritic cells upon recognition of a novel CD99-like ligand by paired immunoglobulin-like type 2 receptor.

Shiratori I, Ogasawara K, Saito T, Lanier LL, Arase H - J. Exp. Med. (2004)

Activation of DCs by PILR-L–expressing cells. 5 × 105 BM-derived DCs were cocultured with PILR-L–transfected 293T cells (closed circles) or parental 293T cells (open circles) at the indicated cell number for 1 d. BM-derived DCs were also stimulated with 1 μg/ml LPS as a positive control. Nitric oxide and mouse TNF-α produced in the culture supernatants were measured. Amounts of nitric oxide and TNF-α produced by DCs cultured in medium only were 1.5 ± 0.1 μM and 48.5 ± 7.7 pg/ml, respectively, and those produced by DCs upon LPS stimulation were 59.1 ± 2.8 μM and 2048 ± 127.9 pg/ml, respectively.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2211832&req=5

fig8: Activation of DCs by PILR-L–expressing cells. 5 × 105 BM-derived DCs were cocultured with PILR-L–transfected 293T cells (closed circles) or parental 293T cells (open circles) at the indicated cell number for 1 d. BM-derived DCs were also stimulated with 1 μg/ml LPS as a positive control. Nitric oxide and mouse TNF-α produced in the culture supernatants were measured. Amounts of nitric oxide and TNF-α produced by DCs cultured in medium only were 1.5 ± 0.1 μM and 48.5 ± 7.7 pg/ml, respectively, and those produced by DCs upon LPS stimulation were 59.1 ± 2.8 μM and 2048 ± 127.9 pg/ml, respectively.
Mentions: BM-DCs were generated by culturing 5 × 106 BM cells with RPMI 1640 medium containing 10% FCS and 200 U/ml mouse GM-CSF for 7–8 d in six-well culture plates. Nonadherent cells were collected and cultured for 1 d. More than 95% of the cells expressed CD11b and CD11c. 5 × 105 BM-DCs were cocultured with various numbers of parental or PILR-L–transfected 293T cells in 48-well plates, as indicated in Fig. 8. After 24 h, culture supernatants were collected, and amounts of NO2 or TNFα were measured by using the Griess reagent (Sigma-Aldrich) or a TNFα ELISA kit (Genzyme), respectively.

Bottom Line: Transcripts of PILR ligand are present in many tissues, including some T cell lines.Cells expressing the PILR ligand specifically activated NK cells and dendritic cells that express the activating PILRbeta.Our findings reveal a new regulatory mechanism of innate immunity by PILR and its CD99-like ligand.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuoku, 260-8670, Japan.

ABSTRACT
Paired receptors that consist of highly related activating and inhibitory receptors are widely involved in the regulation of the immune system. Here, we report a mouse orthologue of the human activating paired immunoglobulin-like type 2 receptor (PILR) beta, which was cloned from a cDNA library of natural killer (NK) cells based on its ability to associate with the DAP12 signaling adaptor protein. The activating PILRbeta was expressed not only on NK cells but also on dendritic cells and macrophages. Furthermore, we have identified a novel CD99-like molecule as a ligand for the activating PILRbeta and inhibitory PILRalpha receptors. Transcripts of PILR ligand are present in many tissues, including some T cell lines. Cells expressing the PILR ligand specifically activated NK cells and dendritic cells that express the activating PILRbeta. Our findings reveal a new regulatory mechanism of innate immunity by PILR and its CD99-like ligand.

Show MeSH