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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: However, in contrast to NK cells, they did not express killer inhibitory receptors, CD16, CD56, or CD57.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 CHO  transfectants specifically. DN invariant Vα24+ T cell clones  (DN2.B9, C6, C7, C9, and D6)  and control CD4+ Vα24+ invariant TCR-negative T cell  clones (SP3.4G9 and 5B2), all at  2 × 105/well were stimulated  with 0.05% glutaraldehyde-fixed CD1d+ CHO transfectants  or control CHO cells (2 × 105/ well). PMA (1 ng/ml) and IL-2  (1 nM) were included, and secreted IL-4 and IFN-γ measured  at 48 h by ELISA in triplicate  (standard deviations shown).  Similar results were obtained  without IL-2. (a) IL-4; (b) IFN-γ.
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Figure 3: Invariant Vα24+ T cells responded to CD1d CHO transfectants specifically. DN invariant Vα24+ T cell clones (DN2.B9, C6, C7, C9, and D6) and control CD4+ Vα24+ invariant TCR-negative T cell clones (SP3.4G9 and 5B2), all at 2 × 105/well were stimulated with 0.05% glutaraldehyde-fixed CD1d+ CHO transfectants or control CHO cells (2 × 105/ well). PMA (1 ng/ml) and IL-2 (1 nM) were included, and secreted IL-4 and IFN-γ measured at 48 h by ELISA in triplicate (standard deviations shown). Similar results were obtained without IL-2. (a) IL-4; (b) IFN-γ.

Mentions: CD1d recognition was assessed initially using CD1d-transfected CHO cells. Each of the five invariant Vα24+ clones assayed responded specifically to the CD1d-transfected CHO cells based upon T cell proliferation (not shown) and cytokine release (Fig. 3). Recognition of CD1d required PMA and mild aldehyde fixation of the target cells, which have been shown in other systems to substitute for certain physiological costimulatory signals (40). Fig. 3 a shows that the CD1d transfectants stimulated IL-4 production from each of the clones except DN2.C9, with the highest levels produced by DN2.B9 and DN2.D6. The CD1d transfectants also strongly stimulated IFN-γ production from three of these clones (DN2.B9, DN2.C9, and DN2.D6) and modest, but specific, IFN-γ release by the other two clones (Fig. 3 b). In contrast, the invariant Vα24− clones did not respond specifically to the CD1d transfectants (Fig. 3, a and b, and data not shown). The failure of the DN2.C9 clone to produce significant IL-4 in response to the CD1d transfectant was consistent with the relatively low ratio of IL-4/IFN-γ produced by this clone in response to anti-CD3 stimulation (Table 3). Indeed, the relative levels of IL-4 and IFN-γ release in response to CD1d from each of the clones were comparable to those observed with anti-CD3 (Table 3).


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 CHO  transfectants specifically. DN invariant Vα24+ T cell clones  (DN2.B9, C6, C7, C9, and D6)  and control CD4+ Vα24+ invariant TCR-negative T cell  clones (SP3.4G9 and 5B2), all at  2 × 105/well were stimulated  with 0.05% glutaraldehyde-fixed CD1d+ CHO transfectants  or control CHO cells (2 × 105/ well). PMA (1 ng/ml) and IL-2  (1 nM) were included, and secreted IL-4 and IFN-γ measured  at 48 h by ELISA in triplicate  (standard deviations shown).  Similar results were obtained  without IL-2. (a) IL-4; (b) IFN-γ.
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Related In: Results  -  Collection

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Figure 3: Invariant Vα24+ T cells responded to CD1d CHO transfectants specifically. DN invariant Vα24+ T cell clones (DN2.B9, C6, C7, C9, and D6) and control CD4+ Vα24+ invariant TCR-negative T cell clones (SP3.4G9 and 5B2), all at 2 × 105/well were stimulated with 0.05% glutaraldehyde-fixed CD1d+ CHO transfectants or control CHO cells (2 × 105/ well). PMA (1 ng/ml) and IL-2 (1 nM) were included, and secreted IL-4 and IFN-γ measured at 48 h by ELISA in triplicate (standard deviations shown). Similar results were obtained without IL-2. (a) IL-4; (b) IFN-γ.
Mentions: CD1d recognition was assessed initially using CD1d-transfected CHO cells. Each of the five invariant Vα24+ clones assayed responded specifically to the CD1d-transfected CHO cells based upon T cell proliferation (not shown) and cytokine release (Fig. 3). Recognition of CD1d required PMA and mild aldehyde fixation of the target cells, which have been shown in other systems to substitute for certain physiological costimulatory signals (40). Fig. 3 a shows that the CD1d transfectants stimulated IL-4 production from each of the clones except DN2.C9, with the highest levels produced by DN2.B9 and DN2.D6. The CD1d transfectants also strongly stimulated IFN-γ production from three of these clones (DN2.B9, DN2.C9, and DN2.D6) and modest, but specific, IFN-γ release by the other two clones (Fig. 3 b). In contrast, the invariant Vα24− clones did not respond specifically to the CD1d transfectants (Fig. 3, a and b, and data not shown). The failure of the DN2.C9 clone to produce significant IL-4 in response to the CD1d transfectant was consistent with the relatively low ratio of IL-4/IFN-γ produced by this clone in response to anti-CD3 stimulation (Table 3). Indeed, the relative levels of IL-4 and IFN-γ release in response to CD1d from each of the clones were comparable to those observed with anti-CD3 (Table 3).

Bottom Line: However, in contrast to NK cells, they did not express killer inhibitory receptors, CD16, CD56, or CD57.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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