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Functionally distinct subsets of CD1d-restricted natural killer T cells revealed by CD1d tetramer staining.

Gumperz JE, Miyake S, Yamamura T, Brenner MB - J. Exp. Med. (2002)

Bottom Line: One subset, which was CD4(-), selectively produced the Th1 cytokines interferon gamma and tumor necrosis factor alpha, and expressed NKG2d, a marker associated with cytolysis of microbially infected and neoplastic cells.Further, for both CD1d-restricted NKT cell subsets, we found that antigenic stimulation induced cytokine production but not perforin expression, whereas exposure to inflammatory factors enhanced perforin expression but did not stimulate cytokine production.These results show that the various activities of CD1d-restricted T cells in tumor rejection, autoimmune disease, and microbial infections could result from activation of functionally distinct subsets, and that inflammatory and antigenic stimuli may influence different effector functions.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital and Harvard Medical School, One Jimmy Fund Way, Boston, MA 02115, USA.

ABSTRACT
CD1d-restricted natural killer (NK)T cells are known to potently secrete T helper (Th)1 and Th2 cytokines and to mediate cytolysis, but it is unclear how these contrasting functional activities are regulated. Using lipid antigen-loaded CD1d tetramers, we have distinguished two subsets of CD1d-restricted T cells in fresh peripheral blood that differ in cytokine production and cytotoxic activation. One subset, which was CD4(-), selectively produced the Th1 cytokines interferon gamma and tumor necrosis factor alpha, and expressed NKG2d, a marker associated with cytolysis of microbially infected and neoplastic cells. This subset up-regulated perforin after exposure to interleukin (IL)-2 or IL-12. In contrast, CD4(+) CD1d-restricted NKT cells potently produced both Th1 and Th2 cytokines, up-regulated perforin in response to stimulation by phorbol myristate acetate and ionomycin but not IL-2 or IL-12, and could be induced to express CD95L. Further, for both CD1d-restricted NKT cell subsets, we found that antigenic stimulation induced cytokine production but not perforin expression, whereas exposure to inflammatory factors enhanced perforin expression but did not stimulate cytokine production. These results show that the various activities of CD1d-restricted T cells in tumor rejection, autoimmune disease, and microbial infections could result from activation of functionally distinct subsets, and that inflammatory and antigenic stimuli may influence different effector functions.

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Flow cytometric probability contour plots showing NKG2d and CD95L staining of CD1d tetramer positive cells. The plots are gated on CD1d tetramer positive lymphocytes. a shows NKG2d staining of a sample before depletion of CD4+ cells (top panel), compared with NKG2d staining after depletion of CD4+ cells (bottom panel). b shows CD4 staining compared with intracellular staining for CD95L for an unstimulated sample (top panel), and a PMA and ionomycin stimulated sample (bottom panel).
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fig5: Flow cytometric probability contour plots showing NKG2d and CD95L staining of CD1d tetramer positive cells. The plots are gated on CD1d tetramer positive lymphocytes. a shows NKG2d staining of a sample before depletion of CD4+ cells (top panel), compared with NKG2d staining after depletion of CD4+ cells (bottom panel). b shows CD4 staining compared with intracellular staining for CD95L for an unstimulated sample (top panel), and a PMA and ionomycin stimulated sample (bottom panel).

Mentions: NKG2d is a lectin encoded in the NK complex that is expressed by NK cells, γδ T cells, and CD8+ αβ T cells, that mediates or costimulates cytolysis of virally and bacterially infected or neoplastic cells that express certain stress-induced antigens (51–54). We investigated CD1d-restricted T cell expression of cell surface NKG2d by two color flow cytometric analysis. As a directly conjugated anti-NKG2d antibody was not available, we compared PBMC samples depleted of CD4+ cells to CD4 undepleted samples to evaluate whether NKG2d expression was biased toward CD4+ or CD4− CD1d-restricted T cells. In PBMC samples that were not depleted of CD4+ cells, approximately half of the CD1d tetramer positive cells stained positively for NKG2d (Fig. 5 a, top panel). In PBMC samples that were CD4 depleted, the fraction of CD1d tetramer-positive cells that were NKG2d-positive was increased (Fig. 5 a, bottom panel), suggesting that the CD4− CD1d-restricted T cell subset is enriched for NKG2d expression compared with the CD4+ subset.


Functionally distinct subsets of CD1d-restricted natural killer T cells revealed by CD1d tetramer staining.

Gumperz JE, Miyake S, Yamamura T, Brenner MB - J. Exp. Med. (2002)

Flow cytometric probability contour plots showing NKG2d and CD95L staining of CD1d tetramer positive cells. The plots are gated on CD1d tetramer positive lymphocytes. a shows NKG2d staining of a sample before depletion of CD4+ cells (top panel), compared with NKG2d staining after depletion of CD4+ cells (bottom panel). b shows CD4 staining compared with intracellular staining for CD95L for an unstimulated sample (top panel), and a PMA and ionomycin stimulated sample (bottom panel).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193772&req=5

fig5: Flow cytometric probability contour plots showing NKG2d and CD95L staining of CD1d tetramer positive cells. The plots are gated on CD1d tetramer positive lymphocytes. a shows NKG2d staining of a sample before depletion of CD4+ cells (top panel), compared with NKG2d staining after depletion of CD4+ cells (bottom panel). b shows CD4 staining compared with intracellular staining for CD95L for an unstimulated sample (top panel), and a PMA and ionomycin stimulated sample (bottom panel).
Mentions: NKG2d is a lectin encoded in the NK complex that is expressed by NK cells, γδ T cells, and CD8+ αβ T cells, that mediates or costimulates cytolysis of virally and bacterially infected or neoplastic cells that express certain stress-induced antigens (51–54). We investigated CD1d-restricted T cell expression of cell surface NKG2d by two color flow cytometric analysis. As a directly conjugated anti-NKG2d antibody was not available, we compared PBMC samples depleted of CD4+ cells to CD4 undepleted samples to evaluate whether NKG2d expression was biased toward CD4+ or CD4− CD1d-restricted T cells. In PBMC samples that were not depleted of CD4+ cells, approximately half of the CD1d tetramer positive cells stained positively for NKG2d (Fig. 5 a, top panel). In PBMC samples that were CD4 depleted, the fraction of CD1d tetramer-positive cells that were NKG2d-positive was increased (Fig. 5 a, bottom panel), suggesting that the CD4− CD1d-restricted T cell subset is enriched for NKG2d expression compared with the CD4+ subset.

Bottom Line: One subset, which was CD4(-), selectively produced the Th1 cytokines interferon gamma and tumor necrosis factor alpha, and expressed NKG2d, a marker associated with cytolysis of microbially infected and neoplastic cells.Further, for both CD1d-restricted NKT cell subsets, we found that antigenic stimulation induced cytokine production but not perforin expression, whereas exposure to inflammatory factors enhanced perforin expression but did not stimulate cytokine production.These results show that the various activities of CD1d-restricted T cells in tumor rejection, autoimmune disease, and microbial infections could result from activation of functionally distinct subsets, and that inflammatory and antigenic stimuli may influence different effector functions.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital and Harvard Medical School, One Jimmy Fund Way, Boston, MA 02115, USA.

ABSTRACT
CD1d-restricted natural killer (NK)T cells are known to potently secrete T helper (Th)1 and Th2 cytokines and to mediate cytolysis, but it is unclear how these contrasting functional activities are regulated. Using lipid antigen-loaded CD1d tetramers, we have distinguished two subsets of CD1d-restricted T cells in fresh peripheral blood that differ in cytokine production and cytotoxic activation. One subset, which was CD4(-), selectively produced the Th1 cytokines interferon gamma and tumor necrosis factor alpha, and expressed NKG2d, a marker associated with cytolysis of microbially infected and neoplastic cells. This subset up-regulated perforin after exposure to interleukin (IL)-2 or IL-12. In contrast, CD4(+) CD1d-restricted NKT cells potently produced both Th1 and Th2 cytokines, up-regulated perforin in response to stimulation by phorbol myristate acetate and ionomycin but not IL-2 or IL-12, and could be induced to express CD95L. Further, for both CD1d-restricted NKT cell subsets, we found that antigenic stimulation induced cytokine production but not perforin expression, whereas exposure to inflammatory factors enhanced perforin expression but did not stimulate cytokine production. These results show that the various activities of CD1d-restricted T cells in tumor rejection, autoimmune disease, and microbial infections could result from activation of functionally distinct subsets, and that inflammatory and antigenic stimuli may influence different effector functions.

Show MeSH
Related in: MedlinePlus