A peptide derived from the Wiskott-Aldrich syndrome (WAS) protein-interacting protein (WIP) restores WAS protein level and actin cytoskeleton reorganization in lymphocytes from patients with WAS mutations that disrupt WIP binding - 11/08/11
, Narayanaswamy Ramesh, PhD a, b, Severine Le Bras, PhD a, b, Silvia Giliani, PhD c, Lucia D. Notarangelo, MD d, Waleed Al-Herz, MD e, Luigi D. Notarangelo, MD a, b, Raif S. Geha, MD a, b
Reprint requests: Michel J. Massaad, PhD, Division of Immunology, Children’s Hospital, Karp Building, 10th Floor, One Blackfan Circle, Boston, MA, 02115.Abstract |
Background |
The Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT) are caused by mutations in WAS, which encodes for WAS protein (WASP). The WASP-interacting protein (WIP) stabilizes WASP, as evidenced by severely decreased WASP levels in T cells from WIP-deficient mice. The majority of missense mutations in patients with WAS/XLT are located in the WIP-binding domain of WASP and might result in dissociation of the WASP-WIP complex and WASP degradation.
Objective |
To restore WASP levels and correct T-cell function in WAS/XLT patients with mutations in the WIP-binding domain of WASP.
Methods |
WIP, and a WIP-derived 41–amino acid–long peptide, which interacts with WASP and was designated nanoWIP (nWIP), were fused to enhanced green fluorescent protein and introduced by electroporation into EBV-transformed B cells, and by retroviral transduction into purified blood T cells from patients with WAS. WASP levels were measured by intracellular fluorescence-activated cell sorting staining. The actin cytoskeleton was visualized by intracellular phalloidin staining.
Results |
Introduction of WIP and nWIP restored WASP levels to normal in EBV-transformed B-cell lines from XLT patients with missense mutations in the WIP-binding domain of WASP and residual WASP levels, and corrected the defective spreading and pseudopodia formation of their T cells in response to immobilized anti-CD3.
Conclusion |
A WASP-binding WIP-derived peptide stabilizes WASP in cells from XLT patients with missense mutations that disrupt WIP binding, and corrects their T-cell actin cytoskeleton defect. This may provide a novel therapeutic strategy for these patients.
El texto completo de este artículo está disponible en PDF.Key words : Wiskott-Aldrich syndrome, WASP, WIP, T cells, actin cytoskeleton
Abbreviations used : aa, EGFP, EBV-B, Ena/VASP, EVH1, MFI, nWIP, TCR, WAS, WASP, WH1, WIP, WT, XLT
Esquema
| Supported by the Perkin fund (M.J.M.), National Institutes of Health grant no. 5PO1HL059561 (R.S.G.), and the Dubai Harvard Foundation for Medical Research. |
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| Disclosure of potential conflict of interest: R. S. Geha has received research support from the National Institutes of Health. The rest of the authors have declared that they have no conflict of interest. |
Vol 127 - N° 4
P. 998 - avril 2011 Regresar al número
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