Related Experiment Videos
The Kit-activating mutation D816V enhances stem cell factor--dependent chemotaxis
M L Taylor1, J Dastych, D Sehgal
1Laboratory of Allergic Diseases and Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1881, USA. mtaylor@niaid.nih.gov
Blood
|August 9, 2001
Summary
The D816V mutation in c-kit enhances the migration of mast cell precursors. This discovery helps explain the abnormal mast cell collections seen in mastocytosis patients.
Area of Science:
- Hematology
- Molecular Biology
- Cell Biology
Background:
- Mastocytosis is characterized by abnormal mast cell accumulation.
- The D816V mutation in c-kit is frequently found in mastocytosis patients.
- This mutation leads to constitutive activation of the Kit tyrosine kinase.
Purpose of the Study:
- To investigate if the D816V c-kit mutation enhances the chemotaxis of mast cell precursors.
- To understand the role of this mutation in the pathophysiology of mastocytosis.
Main Methods:
- Transfection of Jurkat cells with wild-type or D816V mutant Kit.
- Assessment of cell migration towards stem cell factor (SCF) in the presence of tyrosine kinase inhibitors.
- Chemotaxis assays using CD34(+)CD117(+) mast cell precursors from mastocytosis patients.
Main Results:
- D816V mutant Kit significantly enhanced chemotaxis towards SCF compared to wild-type Kit.
- Tyrosine kinase inhibitors reduced migration in both cell types, with D816V transfectants showing higher sensitivity.
- In patient-derived cells, the D816V mutation was significantly enriched in migrated cells compared to pre-migration samples.
Conclusions:
- The D816V c-kit mutation enhances the chemotaxis of CD117(+) cells.
- This enhanced migration provides a potential mechanism for the increased mast cell burden in mastocytosis tissues.