c-Cbl and Cbl-b regulate T cell responsiveness by promoting ligand-induced TCR down-modulation


How Cbl family proteins regulate T cell responses is unclear. We found that c-Cbl Cbl-b double knock-out (dKO) T cells became hyperresponsive upon anti-CD3 stimulation, even though the major T cell antigen receptor (TCR) signaling pathways were not enhanced. The dKO T cells did not down-modulate surface TCR after ligand engagement, which resulted in sustained TCR signaling. However, these cells showed normal ligand-independent TCR internalization, and trafficking of internalized TCR to the lysosome compartment after ligand engagement was reduced. These findings show that Cbl family proteins negatively regulate T cell activation by promoting clearance of engaged TCR from the cell surface, a process that is apparently essential for the termination of TCR signals.

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Figure 1: Expression patterns and targeting strategy for Cbl and Cblb.
Figure 2: General survey of the dKO mice.
Figure 3: Phenotypical analysis and cytokine production profiles of the dKO T cells.
Figure 4: Biochemical analysis of TCR signaling pathways in dKO T cells.
Figure 5: Analysis of TCR down-modulation in the dKO T cells.
Figure 6: Sustained activation of Erk1/2 in the dKO T cells.
Figure 7: Defective ligand-dependent intracellular retention of internalized TCR in dKO T cells.
Figure 8: Impaired lysosomal colocalization of internalized TCRs in the dKO T cells.

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We thank R. N. Germain, W. E. Paul, S. K. Pierce, D. R. Littman and Y. R. Zou for critical review of the manuscript; T. Jin, J. Delon and O. Schwartz for assistance on confocal microscopy analysis; J. Takeda and C. Wilson for Lck-Cre transgenic mice. A. Weiss for anti-CD148; F. Huetz for anti-dsDNA reagents; and L. X. Zheng for 2C11. D. H. is supported by Federal funds from the NCI, NIH, under contract N01-C0-56000.

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Correspondence to Hua Gu.

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Supplementary information

Web Fig 1.

Pathology of the arteritis in the dKO mice. H&E stain of aorta in WT control (a) and dKO (b) mice. Immunohistochemistry of the dKO specimen with anti-B220 (c) and anti-CD3 (d) shows the infiltration of T but not B cells in the vascular wall. (PDF 351 kb)

Web Fig 2.

Block of TCR downmodulation on the c-Cbl–deficient CD4+ CD8+ DP thymocytes. Shown are the histograms of TCR expression on gated CD4+ CD8+ DP thymocytes from the WT and c-Cbl–deficient mice. Cells were stimulated with anti-CD3 for 4 h, and cell surface TCRs were revealed by anti-TCRβ staining. Open and shaded curves represent stimulated and nonstimulated cells, respectively. (PDF 46 kb)

Web Fig 3.

Defective ligand-dependent intracellular TCR retention in dKO T cells. Intracellular retention of internalized TCR was detected as described in Methods. Open and shaded curves represent cells before and after the acidic buffer treatment, respectively. (PDF 113 kb)

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Naramura, M., Jang, I., Kole, H. et al. c-Cbl and Cbl-b regulate T cell responsiveness by promoting ligand-induced TCR down-modulation. Nat Immunol 3, 1192–1199 (2002). https://doi.org/10.1038/ni855

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