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Leukocyte adhesion deficiency-III is caused by mutations in KINDLIN3 affecting integrin activation

Nature Medicine volume 15, pages 306–312 (2009)Cite this article

Abstract

Integrins are the major adhesion receptors of leukocytes and platelets. β1 and β2 integrin function on leukocytes is crucial for a successful immune response and the platelet integrin αIIbβ3 initiates the process of blood clotting through binding fibrinogen1,2,3. Integrins on circulating cells bind poorly to their ligands but become active after 'inside-out' signaling through other membrane receptors4,5. Subjects with leukocyte adhesion deficiency-1 (LAD-I) do not express β2 integrins because of mutations in the gene specifying the β2 subunit, and they suffer recurrent bacterial infections6,7. Mutations affecting αIIbβ3 integrin cause the bleeding disorder termed Glanzmann's thrombasthenia3. Subjects with LAD-III show symptoms of both LAD-I and Glanzmann's thrombasthenia. Their hematopoietically-derived cells express β1, β2 and β3 integrins, but defective inside-out signaling causes immune deficiency and bleeding problems8. The LAD-III lesion has been attributed to a C → A mutation in the gene encoding calcium and diacylglycerol guanine nucleotide exchange factor (CALDAGGEF1; official symbol RASGRP2) specifying the CALDAG-GEF1 protein9, but we show that this change is not responsible for the LAD-III disorder. Instead, we identify mutations in the KINDLIN3 (official symbol FERMT3) gene specifying the KINDLIN-3 protein as the cause of LAD-III in Maltese and Turkish subjects. Two independent mutations result in decreased KINDLIN3 messenger RNA levels and loss of protein expression. Notably, transfection of the subjects' lymphocytes with KINDLIN3 complementary DNA but not CALDAGGEF1 cDNA reverses the LAD-III defect, restoring integrin-mediated adhesion and migration.

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Figure 1: The CALDAGGEF1 gene C → A base change has no effect on mRNA and protein levels or on deficient LAD-III B cell adhesion and migration.
Figure 2: Mutations in the KINDLIN3 gene.
Figure 3: Analysis of mutated KINDLIN3 mRNA in subjects with LAD-III and their relatives.
Figure 4: Adhesion and migration characteristics of LAD-III B cells expressing wild-type Kindlin-3.

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Acknowledgements

We are indebted to R. Fässler and D. Wagner for helpful discussion; J. Crittenden and A. Graybiel (Massachusetts Institute of Technology) for antibodies to CALDAG-GEF1, cDNA constructs and helpful discussion; and J. Hancock (University of Queensland Medical School) for CALDAGGEF1 constructs. We are also grateful to our Cancer Research UK London Research Institute colleagues D. Harvey for generation of the EBV-transformed cell lines and G. Kelly for help with the statistical analyses. L. S. was supported by a Marie Curie Individual Fellowship.

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Author notes

  1. Lena Svensson and Kimberley Howarth: These authors contributed equally to this work.

Authors and Affiliations

  1. Leukocyte Adhesion Laboratory, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Lena Svensson, Alison McDowall, Irene Patzak, Rachel Evans & Nancy Hogg

  2. Molecular and Population Genetics Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Kimberley Howarth & Ian Tomlinson

  3. Department of Molecular Medicine, Max Planck Institute of Biochemistry, Am Klopferspitz 18, Martinsried, 82152, Germany

    Siegfried Ussar & Markus Moser

  4. Division of Immunology, SB Ankara Diskapi Children's Hospital, Ankara, 06110, Turkey

    Ayse Metin

  5. University of California–San Francisco Cancer Research Institute, 2340 Sutter Street, San Francisco, 94115, California, USA

    Mike Fried

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Contributions

L.S., K.H., I.T., M.F. and N.H. designed the experiments; L.S., K.H., A.M., I.P. and R.E. performed the experiments; and A.M. provided subject blood samples; S.U. provided the antibody to Kindlin-3; M.M. provided the EGFP–Kindlin-3 construct;. L.S., K.H., A.M., I.P., M.F., I.T. and N.H. were involved in data analyses. All authors contributed to the writing or editing of the manuscript. N.H. supervised the project and wrote the initial manuscript.

Corresponding author

Correspondence to Nancy Hogg.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–5 and Supplementary Methods (PDF 5122 kb)

Supplementary Video 1

GFP-expressing LAD-III B cells (Turkish family three) migrating on ICAM-1. (MOV 7433 kb)

Supplementary Video 2

CALDAG-GEF-1–expressing LAD-III B cells (Turkish family three) migrating on ICAM-1. (MOV 4545 kb)

Supplementary Video 3

Kindlin-3–expressing LAD-III B cells (Turkish family three) migrating on ICAM-1. (MOV 8949 kb)

Supplementary Video 4

Kindlin-3–expressing LAD-III B cells (Turkish family three) migrating on ICAM-1. (MOV 9724 kb)

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Svensson, L., Howarth, K., McDowall, A. et al. Leukocyte adhesion deficiency-III is caused by mutations in KINDLIN3 affecting integrin activation. Nat Med 15, 306–312 (2009). https://doi.org/10.1038/nm.1931

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