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Cross-presentation of disialoganglioside GD3 to natural killer T cells.

Wu DY, Segal NH, Sidobre S, Kronenberg M, Chapman PB - J. Exp. Med. (2003)

Bottom Line: GD3-reactive NKT cells initially produced IL-4 and IFN-gamma followed by IL-10.Because SK-MEL-28 does not express any isoform of human CD1, GD3 must be cross-presented by murine APCs in vivo.This could be a mechanism for NKT cell recognition of tumor gangliosides in CD1- tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

ABSTRACT
GD3, a ganglioside expressed on human melanoma, can be recognized by the humoral immune system. In this paper, we demonstrate that immunizing mice with the human melanoma cell line SK-MEL-28 (GD3+ GM2- CD1-) or with syngeneic APCs loaded with GD3 can induce a GD3-reactive natural killer T (NKT) cell response. GD3-reactive NKT cells were detected among splenocytes of immunized mice at frequencies of approximately 1:2000 both by ELISPOT and GD3-loaded mouse CD1d tetramer analysis. GD3-reactive NKT cells did not react with GM2, a closely related ganglioside, and were not detectable in unimmunized mice. GD3-reactive NKT cells initially produced IL-4 and IFN-gamma followed by IL-10. They were CD1d restricted in that reactivity was abrogated when APCs were blocked with anti-CD1d monoclonal antibody before being loaded with GD3 or when APCs from CD1d knockout mice were used. Because SK-MEL-28 does not express any isoform of human CD1, GD3 must be cross-presented by murine APCs in vivo. This is the first analysis of a natural ligand for mouse NKT cells and the first definitive paper of cross-presentation to NKT cells. This could be a mechanism for NKT cell recognition of tumor gangliosides in CD1- tumors.

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CD1d expression by APCs is required for presentation of GD3 to NKT cells. Splenocytes from mice immunized with SK-MEL-28 and Freund's adjuvant were used as effector cells and IL-4 production by splenocytes was detected by ELISPOT assay. APCs were isolated from the spleen of mCD1d knockout mice (CD1−/−) or from wild-type mice (CD1+/+) as indicated and loaded with GD3. Control wells received unloaded APCs. Each point represents the mean of triplicate wells from an individual mouse; horizontal lines indicate mean values.
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fig5: CD1d expression by APCs is required for presentation of GD3 to NKT cells. Splenocytes from mice immunized with SK-MEL-28 and Freund's adjuvant were used as effector cells and IL-4 production by splenocytes was detected by ELISPOT assay. APCs were isolated from the spleen of mCD1d knockout mice (CD1−/−) or from wild-type mice (CD1+/+) as indicated and loaded with GD3. Control wells received unloaded APCs. Each point represents the mean of triplicate wells from an individual mouse; horizontal lines indicate mean values.

Mentions: To confirm that GD3 recognition by NKT cells was CD1d dependent, splenocytes from immunized mice were stimulated with GD3-loaded APCs from CD1d knockout (CD1d−/−) or wild-type (CD1d+/+) mice. As shown in Fig. 5, GD3-loaded APCs derived from CD1−/− mice were not able to present GD3 to GD3-reactive NKTs. These results confirm that CD1d is required for presentation of GD3 to the NKTs.


Cross-presentation of disialoganglioside GD3 to natural killer T cells.

Wu DY, Segal NH, Sidobre S, Kronenberg M, Chapman PB - J. Exp. Med. (2003)

CD1d expression by APCs is required for presentation of GD3 to NKT cells. Splenocytes from mice immunized with SK-MEL-28 and Freund's adjuvant were used as effector cells and IL-4 production by splenocytes was detected by ELISPOT assay. APCs were isolated from the spleen of mCD1d knockout mice (CD1−/−) or from wild-type mice (CD1+/+) as indicated and loaded with GD3. Control wells received unloaded APCs. Each point represents the mean of triplicate wells from an individual mouse; horizontal lines indicate mean values.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196074&req=5

fig5: CD1d expression by APCs is required for presentation of GD3 to NKT cells. Splenocytes from mice immunized with SK-MEL-28 and Freund's adjuvant were used as effector cells and IL-4 production by splenocytes was detected by ELISPOT assay. APCs were isolated from the spleen of mCD1d knockout mice (CD1−/−) or from wild-type mice (CD1+/+) as indicated and loaded with GD3. Control wells received unloaded APCs. Each point represents the mean of triplicate wells from an individual mouse; horizontal lines indicate mean values.
Mentions: To confirm that GD3 recognition by NKT cells was CD1d dependent, splenocytes from immunized mice were stimulated with GD3-loaded APCs from CD1d knockout (CD1d−/−) or wild-type (CD1d+/+) mice. As shown in Fig. 5, GD3-loaded APCs derived from CD1−/− mice were not able to present GD3 to GD3-reactive NKTs. These results confirm that CD1d is required for presentation of GD3 to the NKTs.

Bottom Line: GD3-reactive NKT cells initially produced IL-4 and IFN-gamma followed by IL-10.Because SK-MEL-28 does not express any isoform of human CD1, GD3 must be cross-presented by murine APCs in vivo.This could be a mechanism for NKT cell recognition of tumor gangliosides in CD1- tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

ABSTRACT
GD3, a ganglioside expressed on human melanoma, can be recognized by the humoral immune system. In this paper, we demonstrate that immunizing mice with the human melanoma cell line SK-MEL-28 (GD3+ GM2- CD1-) or with syngeneic APCs loaded with GD3 can induce a GD3-reactive natural killer T (NKT) cell response. GD3-reactive NKT cells were detected among splenocytes of immunized mice at frequencies of approximately 1:2000 both by ELISPOT and GD3-loaded mouse CD1d tetramer analysis. GD3-reactive NKT cells did not react with GM2, a closely related ganglioside, and were not detectable in unimmunized mice. GD3-reactive NKT cells initially produced IL-4 and IFN-gamma followed by IL-10. They were CD1d restricted in that reactivity was abrogated when APCs were blocked with anti-CD1d monoclonal antibody before being loaded with GD3 or when APCs from CD1d knockout mice were used. Because SK-MEL-28 does not express any isoform of human CD1, GD3 must be cross-presented by murine APCs in vivo. This is the first analysis of a natural ligand for mouse NKT cells and the first definitive paper of cross-presentation to NKT cells. This could be a mechanism for NKT cell recognition of tumor gangliosides in CD1- tumors.

Show MeSH
Related in: MedlinePlus