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Requirements for CD1d recognition by human invariant Valpha24+ CD4-CD8- T cells.

Exley M, Garcia J, Balk SP, Porcelli S - J. Exp. Med. (1997)

Bottom Line: Upon activation by anti-CD3 or CD1d, the clones produced both Th1 and Th2 cytokines.These results demonstrate that human invariant Valpha24+ CD4-CD8- T cells, and presumably the homologous murine NK1+ T cell population, are CD1d reactive and functionally distinct from NK cells.The conservation of this cell population and of the CD1d ligand across species indicates an important immunological function.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology Program, Hematology/Oncology Division, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
A subset of human CD4-CD8- T cells that expresses an invariant Valpha24-JalphaQ T cell receptor (TCR)-alpha chain, paired predominantly with Vbeta11, has been identified. A series of these Valpha24 Vbeta11 clones were shown to have TCR-beta CDR3 diversity and express the natural killer (NK) locus-encoded C-type lectins NKR-P1A, CD94, and CD69. However, in contrast to NK cells, they did not express killer inhibitory receptors, CD16, CD56, or CD57. All invariant Valpha24(+) clones recognized the MHC class I-like CD16 molecule and discriminated between CD1d and other closely related human CD1 proteins, indicating that recognition was TCR-mediated. Recognition was not dependent upon an endosomal targeting motif in the cytoplasmic tail of CD1d. Upon activation by anti-CD3 or CD1d, the clones produced both Th1 and Th2 cytokines. These results demonstrate that human invariant Valpha24+ CD4-CD8- T cells, and presumably the homologous murine NK1+ T cell population, are CD1d reactive and functionally distinct from NK cells. The conservation of this cell population and of the CD1d ligand across species indicates an important immunological function.

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Invariant Vα24+ T  cells responded to CD1d B cell  transfectants. T cell clones (105/ well) were incubated for 48 h  with either fixed (0.025 or 0.05%  glutaraldehyde, latter shown) or  unfixed C1R human B cells  (105/well) transfected with CD1a,  b, c, d, or plasmid alone. Results  with the two fixations were indistinguishable. PMA (1 ng/ml) was  included and PHA as positive  control is shown for comparison.  Representative IFN-γ cytokine  ELISA results of multiple experiments are shown. (a) Production  of IFN-γ by DN2.C9 incubated  for 48 h with C1R ± fixation.  (b) IFN-γ cytokine responses of  three representative DN2 T cell  clones incubated for 48 h with  unfixed C1R transfected with  CD1a, b, c, d, mock, or mock  with PHA.
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Figure 5: Invariant Vα24+ T cells responded to CD1d B cell transfectants. T cell clones (105/ well) were incubated for 48 h with either fixed (0.025 or 0.05% glutaraldehyde, latter shown) or unfixed C1R human B cells (105/well) transfected with CD1a, b, c, d, or plasmid alone. Results with the two fixations were indistinguishable. PMA (1 ng/ml) was included and PHA as positive control is shown for comparison. Representative IFN-γ cytokine ELISA results of multiple experiments are shown. (a) Production of IFN-γ by DN2.C9 incubated for 48 h with C1R ± fixation. (b) IFN-γ cytokine responses of three representative DN2 T cell clones incubated for 48 h with unfixed C1R transfected with CD1a, b, c, d, mock, or mock with PHA.

Mentions: Stimulation of invariant Vα24+ T cells by CD1d+ CHO cells required both PMA and aldehyde fixation, presumably due to the absence of necessary costimulatory ligands on the CHO cells. Although the target cells that mediate activation of invariant Vα24+ T cells in vivo are not known, normal B cells express CD1d (51) and may be a relevant CD1d-presenting cell. Therefore, the C1R HLA-A and -B negative B lymphoblastoid cell line (52), which does not express detectable CD1d (our unpublished data), was used to confirm the results in CHO cells and to determine whether the need for nonphysiological costimulation could be reduced or eliminated. CD1d-transfected C1R cells specifically stimulated each of the invariant Vα24+ DN T cell clones tested based upon IFN-γ and IL-4 production and T cell proliferation (Fig. 5 a and data not shown), confirming the results in CHO cells. Stimulation did not require aldehyde fixation, but phorbol ester was still necessary.


Requirements for CD1d recognition by human invariant Valpha24+ CD4-CD8- T cells.

Exley M, Garcia J, Balk SP, Porcelli S - J. Exp. Med. (1997)

Invariant Vα24+ T  cells responded to CD1d B cell  transfectants. T cell clones (105/ well) were incubated for 48 h  with either fixed (0.025 or 0.05%  glutaraldehyde, latter shown) or  unfixed C1R human B cells  (105/well) transfected with CD1a,  b, c, d, or plasmid alone. Results  with the two fixations were indistinguishable. PMA (1 ng/ml) was  included and PHA as positive  control is shown for comparison.  Representative IFN-γ cytokine  ELISA results of multiple experiments are shown. (a) Production  of IFN-γ by DN2.C9 incubated  for 48 h with C1R ± fixation.  (b) IFN-γ cytokine responses of  three representative DN2 T cell  clones incubated for 48 h with  unfixed C1R transfected with  CD1a, b, c, d, mock, or mock  with PHA.
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Related In: Results  -  Collection

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Figure 5: Invariant Vα24+ T cells responded to CD1d B cell transfectants. T cell clones (105/ well) were incubated for 48 h with either fixed (0.025 or 0.05% glutaraldehyde, latter shown) or unfixed C1R human B cells (105/well) transfected with CD1a, b, c, d, or plasmid alone. Results with the two fixations were indistinguishable. PMA (1 ng/ml) was included and PHA as positive control is shown for comparison. Representative IFN-γ cytokine ELISA results of multiple experiments are shown. (a) Production of IFN-γ by DN2.C9 incubated for 48 h with C1R ± fixation. (b) IFN-γ cytokine responses of three representative DN2 T cell clones incubated for 48 h with unfixed C1R transfected with CD1a, b, c, d, mock, or mock with PHA.
Mentions: Stimulation of invariant Vα24+ T cells by CD1d+ CHO cells required both PMA and aldehyde fixation, presumably due to the absence of necessary costimulatory ligands on the CHO cells. Although the target cells that mediate activation of invariant Vα24+ T cells in vivo are not known, normal B cells express CD1d (51) and may be a relevant CD1d-presenting cell. Therefore, the C1R HLA-A and -B negative B lymphoblastoid cell line (52), which does not express detectable CD1d (our unpublished data), was used to confirm the results in CHO cells and to determine whether the need for nonphysiological costimulation could be reduced or eliminated. CD1d-transfected C1R cells specifically stimulated each of the invariant Vα24+ DN T cell clones tested based upon IFN-γ and IL-4 production and T cell proliferation (Fig. 5 a and data not shown), confirming the results in CHO cells. Stimulation did not require aldehyde fixation, but phorbol ester was still necessary.

Bottom Line: Upon activation by anti-CD3 or CD1d, the clones produced both Th1 and Th2 cytokines.These results demonstrate that human invariant Valpha24+ CD4-CD8- T cells, and presumably the homologous murine NK1+ T cell population, are CD1d reactive and functionally distinct from NK cells.The conservation of this cell population and of the CD1d ligand across species indicates an important immunological function.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology Program, Hematology/Oncology Division, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
A subset of human CD4-CD8- T cells that expresses an invariant Valpha24-JalphaQ T cell receptor (TCR)-alpha chain, paired predominantly with Vbeta11, has been identified. A series of these Valpha24 Vbeta11 clones were shown to have TCR-beta CDR3 diversity and express the natural killer (NK) locus-encoded C-type lectins NKR-P1A, CD94, and CD69. However, in contrast to NK cells, they did not express killer inhibitory receptors, CD16, CD56, or CD57. All invariant Valpha24(+) clones recognized the MHC class I-like CD16 molecule and discriminated between CD1d and other closely related human CD1 proteins, indicating that recognition was TCR-mediated. Recognition was not dependent upon an endosomal targeting motif in the cytoplasmic tail of CD1d. Upon activation by anti-CD3 or CD1d, the clones produced both Th1 and Th2 cytokines. These results demonstrate that human invariant Valpha24+ CD4-CD8- T cells, and presumably the homologous murine NK1+ T cell population, are CD1d reactive and functionally distinct from NK cells. The conservation of this cell population and of the CD1d ligand across species indicates an important immunological function.

Show MeSH