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

Nature Immunology volume 3, pages 1192–1199 (2002)Cite this article

Abstract

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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Acknowledgements

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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Author notes

  1. Mayumi Naramura

    Present address: Department of Immunology, Cleveland Clinic Foundation, NB-30, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

  2. Mayumi Naramura, Ihn-Kyung Jang and Hemanta Kole: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, 20852, MD, USA

    Mayumi Naramura, Ihn-Kyung Jang, Hemanta Kole, Fang Huang & Hua Gu

  2. Pathology/Histotechnology Laboratory, National Cancer Institute-Frederick Cancer Research & Development Center, Frederick, 21702, MD, USA

    Diana Haines

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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, IK., 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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