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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 that did not express Vβ11  responded to CD1d. (a) Representative FACS® profiles of  two DN invariant TCR+ T cell  lines (DN2.Vβ11+, left; and  DN2.Vβ11−, right). T cells  were stained with 10 μg/ml of  mAbs against Vα24, Vβ11, or  isotype control mAb and with  anti-IgG FITC conjugate for 30  min each before FACS® analysis  of viable cells. (b) T cell lines  (105/well) were incubated with  unfixed C1R human B cells  (105/well) transfected with CD1a,  b, c, d, mock, or mock with  PHA as in Fig. 5 and IFN-γ production results are shown.
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Figure 6: Invariant Vα24+ T cells that did not express Vβ11 responded to CD1d. (a) Representative FACS® profiles of two DN invariant TCR+ T cell lines (DN2.Vβ11+, left; and DN2.Vβ11−, right). T cells were stained with 10 μg/ml of mAbs against Vα24, Vβ11, or isotype control mAb and with anti-IgG FITC conjugate for 30 min each before FACS® analysis of viable cells. (b) T cell lines (105/well) were incubated with unfixed C1R human B cells (105/well) transfected with CD1a, b, c, d, mock, or mock with PHA as in Fig. 5 and IFN-γ production results are shown.

Mentions: Polyclonal DN T cell lines selected for expression of Vα24 were sorted into Vβ11+ or Vβ11− populations and examined to determine whether Vβ11 was necessary for CD1d recognition. FACS® analyses showed that virtually all of the cells in both lines were Vα24+ (Fig. 6 a), and previous RT-PCR analyses of these lines showed that both expressed primarily or exclusively the invariant Vα24 (32). The line designated as DN2.Vβ11− had no significant Vβ11+ population, whereas the line designated as DN2.Vβ11+ was virtually all Vβ11+ (Fig. 6 a). It is of interest that cells in the Vβ11− line consistently expressed slightly lower TCR levels, based upon staining with the anti-Vα24 mAb (Fig. 6 a) and anti-TCR mAbs (not shown). The relationship of this observation to the preferential use of Vβ11 by invariant Vα24+ T cells is not clear, but could reflect greater stability of the invariant Vα24 when paired with Vβ11.


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 that did not express Vβ11  responded to CD1d. (a) Representative FACS® profiles of  two DN invariant TCR+ T cell  lines (DN2.Vβ11+, left; and  DN2.Vβ11−, right). T cells  were stained with 10 μg/ml of  mAbs against Vα24, Vβ11, or  isotype control mAb and with  anti-IgG FITC conjugate for 30  min each before FACS® analysis  of viable cells. (b) T cell lines  (105/well) were incubated with  unfixed C1R human B cells  (105/well) transfected with CD1a,  b, c, d, mock, or mock with  PHA as in Fig. 5 and IFN-γ production results are shown.
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Related In: Results  -  Collection

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Figure 6: Invariant Vα24+ T cells that did not express Vβ11 responded to CD1d. (a) Representative FACS® profiles of two DN invariant TCR+ T cell lines (DN2.Vβ11+, left; and DN2.Vβ11−, right). T cells were stained with 10 μg/ml of mAbs against Vα24, Vβ11, or isotype control mAb and with anti-IgG FITC conjugate for 30 min each before FACS® analysis of viable cells. (b) T cell lines (105/well) were incubated with unfixed C1R human B cells (105/well) transfected with CD1a, b, c, d, mock, or mock with PHA as in Fig. 5 and IFN-γ production results are shown.
Mentions: Polyclonal DN T cell lines selected for expression of Vα24 were sorted into Vβ11+ or Vβ11− populations and examined to determine whether Vβ11 was necessary for CD1d recognition. FACS® analyses showed that virtually all of the cells in both lines were Vα24+ (Fig. 6 a), and previous RT-PCR analyses of these lines showed that both expressed primarily or exclusively the invariant Vα24 (32). The line designated as DN2.Vβ11− had no significant Vβ11+ population, whereas the line designated as DN2.Vβ11+ was virtually all Vβ11+ (Fig. 6 a). It is of interest that cells in the Vβ11− line consistently expressed slightly lower TCR levels, based upon staining with the anti-Vα24 mAb (Fig. 6 a) and anti-TCR mAbs (not shown). The relationship of this observation to the preferential use of Vβ11 by invariant Vα24+ T cells is not clear, but could reflect greater stability of the invariant Vα24 when paired with Vβ11.

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
Related in: MedlinePlus