Characterization of a novel human mast cell line that responds to stem cell factor and expresses functional FcεRI - 10/08/11
Abstract |
Background |
Studies of human mast cells (MCs) are constrained by the paucity of functional cell lines, the expense of maintaining MCs in culture, and technical complexities.
Objective |
We derived and characterized a human MC line that arose spontaneously from a culture of nontransformed hematopoietic progenitor cells.
Methods |
CD34+ enriched mononuclear cells derived from a donor with aspirin-exacerbated respiratory disease were cultured for 8 weeks with stem cell factor and IL-6 and with IL-3 for the first week only. The cells (termed LUVA cells) survived and proliferated without further addition of any growth factors and have been maintained in culture for approximately 2 years.
Results |
LUVA cells possess metachromatic cytoplasmic granules that are immunoreactive for tryptase, cathepsin G, and carboxypeptidase A3. They express transcripts encoding FcεRI, c-kit, chymase, tryptase, histidine decarboxylase, carboxypeptidase A3, and the type 1 receptor for cysteinyl leukotrienes. Flow cytometry confirmed uniform expression of FcεRI, c-kit, and FcγRII. FcεRI cross-linkage induced the release of β-hexosaminidase, prostaglandin D2, thromboxane A2, and macrophage inflammatory protein 1β. Immortalization was not associated with either a known genomic mutation of c-kit in the donor or a somatic mutation of c-kit within the cells, and it was not associated with c-kit autophosphorylation.
Conclusions |
LUVA cells are an immortalized human MC line that can be maintained without stem cell factor and display high levels of normally signaling c-kit and FcεRI. These cells will prove valuable for functional human MC studies.
Le texte complet de cet article est disponible en PDF.Key words : Mast cell, stem cell factor, FcεRI, c-kit, tryptase, carboxypeptidase A3
Abbreviations used : CPA3, CREB, cys-LT, ERK, LTC4, LTC4S, MC, MEK, MIP-1β, PGD2, qPCR, SCF, TXA2
Plan
Supported by National Institutes of Health grants AI-57438, AI-50989, AI-52353, AI-31599, AI 07306, and HL-36110; by a grant from the Charles A. Dana Foundation; and by generous contributions from the Vinik Family. T.M.L. was supported by an American Academy of Allergy, Asthma & Immunology third- and fourth-year Fellow’s Award. |
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Disclosure of potential conflict of interest: J. W. Steinke has received research support from the National Institutes of Health. J. A. Boyce has received research support from the National Institutes of Health and Merck. L. Borish has consulted for Hoffman LaRoche, Cephalon, Regeneron, and Pfizer; has received honoraria and research support from Merck; and has received research support from Genentech. The rest of the authors have declared that they have no conflict of interest. |
Vol 127 - N° 3
P. 815 - mars 2011 Retour au numéroBienvenue sur EM-consulte, la référence des professionnels de santé.
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