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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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The phenotype of  thymus and spleen rat DC. DC  were purified as described in the  Materials and Methods section.  Expression of the indicated antigens was analyzed in single color  immunofluorescence. Propidium iodide was used to exclude  dead cells from analysis. Dotted  lines show the background with  isotype-matched control antibody. Each staining was performed at least six times with  similar results.
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Figure 1: The phenotype of thymus and spleen rat DC. DC were purified as described in the Materials and Methods section. Expression of the indicated antigens was analyzed in single color immunofluorescence. Propidium iodide was used to exclude dead cells from analysis. Dotted lines show the background with isotype-matched control antibody. Each staining was performed at least six times with similar results.

Mentions: Using the protocol described in the Materials and Methods section, we routinely obtained spleen and thymus DC populations 95– 98% homogeneous. These cells exhibit a typical dendritic morphology (data not shown) and lack of esterase and myeloperoxidase (data not shown), and express very high levels of class II and B7 molecules (Fig. 1). They do not express T or B cell receptors, sialoadhesin (Fig. 1), nor CD8 (Fig. 2). Moreover, they express high levels of CD25, and the antigen recognized by the OX62 mAb was expressed at a low to moderate level by a subset of spleen and thymus DC (Fig. 1). The complete phenotype and morphological features of rat DC will be described elsewhere (our manuscript in preparation). DC preparations from both thymus and spleen had potent stimulatory activity (>300-fold superior to total spleen cells) in primary allogeneic mixed leukocyte culture (data not shown). Of particular interest was that >90% of spleen and thymus DC expressed the NKR-P1 molecule recognized by the 3.2.3 mAb (Fig. 1). DC expression of NKR-P1 was not strain-restricted since we have observed similar staining for this molecule on DC isolated from Lewis, Wistar, and Sprague-Dawley rats. In contrast, spleen DC from the C57Bl/6 mouse strain do not express the NKR-P1C molecule (23).


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)

The phenotype of  thymus and spleen rat DC. DC  were purified as described in the  Materials and Methods section.  Expression of the indicated antigens was analyzed in single color  immunofluorescence. Propidium iodide was used to exclude  dead cells from analysis. Dotted  lines show the background with  isotype-matched control antibody. Each staining was performed at least six times with  similar results.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: The phenotype of thymus and spleen rat DC. DC were purified as described in the Materials and Methods section. Expression of the indicated antigens was analyzed in single color immunofluorescence. Propidium iodide was used to exclude dead cells from analysis. Dotted lines show the background with isotype-matched control antibody. Each staining was performed at least six times with similar results.
Mentions: Using the protocol described in the Materials and Methods section, we routinely obtained spleen and thymus DC populations 95– 98% homogeneous. These cells exhibit a typical dendritic morphology (data not shown) and lack of esterase and myeloperoxidase (data not shown), and express very high levels of class II and B7 molecules (Fig. 1). They do not express T or B cell receptors, sialoadhesin (Fig. 1), nor CD8 (Fig. 2). Moreover, they express high levels of CD25, and the antigen recognized by the OX62 mAb was expressed at a low to moderate level by a subset of spleen and thymus DC (Fig. 1). The complete phenotype and morphological features of rat DC will be described elsewhere (our manuscript in preparation). DC preparations from both thymus and spleen had potent stimulatory activity (>300-fold superior to total spleen cells) in primary allogeneic mixed leukocyte culture (data not shown). Of particular interest was that >90% of spleen and thymus DC expressed the NKR-P1 molecule recognized by the 3.2.3 mAb (Fig. 1). DC expression of NKR-P1 was not strain-restricted since we have observed similar staining for this molecule on DC isolated from Lewis, Wistar, and Sprague-Dawley rats. In contrast, spleen DC from the C57Bl/6 mouse strain do not express the NKR-P1C molecule (23).

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