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Adaptor protein 3–dependent microtubule-mediated movement of lytic granules to the immunological synapse

Nature Immunology volume 4, pages 1111–1120 (2003)Cite this article

Abstract

Hermansky-Pudlak syndrome (HPS) is a rare autosomal recessive disease characterized by platelet defects and oculocutaneous albinism. Individuals with HPS type 2 (HPS2) lack the cytosolic adaptor protein 3 (AP-3) involved in lysosomal sorting, and are also immunodeficient. Here we characterize an HPS2 mutation and demonstrate that AP-3 deficiency leads to a loss of cytotoxic T lymphocyte (CTL)-mediated cytotoxicity. Although the lysosomal protein CD63 was mislocalized to the plasma membrane, perforin and granzymes were correctly localized to the lytic granules in AP-3-deficient CTLs. However, the lytic granules of AP-3-deficient CTLs were enlarged and were unable to move along microtubules and dock within the secretory domain of the immunological synapse. These data show that AP-3 is essential for polarized secretion from CTLs.

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Figure 1: The AP-3 complex is unstable in HPS2 CTLs.
Figure 2: Identification of the mutations giving rise to HPS2.
Figure 3: CD63 is mis-sorted in AP-3-deficient CTLs.
Figure 4: AP-3-deficient CTLs show a reduced ability to kill or to secrete.
Figure 5: Lytic proteins are correctly localized in AP-3-deficient CTLs.
Figure 6: The lytic granules of AP-3 deficient CTLs are enlarged and show an abnormal distribution in the cell.
Figure 7: Lytic granules from AP-3-deficient CTLs do not polarize properly at the immunological synapse.

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Acknowledgements

We thank J. Kaufman, F. Gallo and N. Barclay for critical reading of the manuscript, and P. Klenerman and his laboratory for providing some reagents. We also thank the individual who made this study possible. Supported by the Wellcome Trust.

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Author notes

  1. Richard H Clark and Jane C Stinchcombe: These authors contributed equally to this work.

Authors and Affiliations

  1. Sir William Dunn School of Pathology, South Parks Road, Oxford, OX1 3RE, UK

    Richard H Clark, Jane C Stinchcombe, Anna Day, Emma Blott, Sarah Booth, Giovanna Bossi & Gillian M Griffiths

  2. Department of Haematology, Royal Bournemouth Hospital, Bournemouth, BH7 7DW, UK

    Terry Hamblin

  3. Great Ormond Street Hospital for Children, London, WC1N 3GH, UK

    E Graham Davies

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Supplementary information

Supplementary Video 1.

Wild-type CTL without target. 3D reconstruction of Fig 7c. (GIF 1061 kb)

Supplementary Video 2.

Wild-type CTL with target. 3D reconstruction of Fig 7d. (GIF 572 kb)

Supplementary Video 3.

AP-3-/- CTL without target. 3D reconstruction of Fig 7e. (GIF 958 kb)

Supplementary Video 4.

AP-3 -/- CTL with target. 3D reconstruction of Fig 7f. (GIF 450 kb)

Supplementary Video 5.

Lytic granule movement in wild-type CTL. (GIF 2007 kb)

Supplementary Video 6.

Lytic granule movement in AP-3 deficient CTL. (GIF 3078 kb)

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Clark, R., Stinchcombe, J., Day, A. et al. Adaptor protein 3–dependent microtubule-mediated movement of lytic granules to the immunological synapse. Nat Immunol 4, 1111–1120 (2003). https://doi.org/10.1038/ni1000

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