title: Demonstration that mast cells, T cells, and B cells bearing the activating kit mutation D816V occur in clusters within the marrow of patients with mastocytosis.
abstract: Mastocytosis is characterized by focal heterotypic clusters of mast cells and lymphocytes in the bone marrow and by a somatically acquired activating Kit mutation, D816V. The relationship of the occurrence of this mutation to the heterotypic clusters of mast cells and lymphocytes in bone marrow is unknown. We hypothesized that these two unique features of mastocytosis were related. To explore this hypothesis, laser capture microdissected mast cells, B cells, and T cells, from both lesional and non-lesional areas of bone marrow biopsy tissues from patients with mastocytosis, were examined for the D816V mutation in their DNA, using HinfI restriction digestion of nested PCR products amplified from extracts of dissected cells. The D816V mutation was detected in mast cells, B cells, and T cells from lesional but not non-lesional areas of bone marrow tissues. B cells obtained from lesional areas of tissue were also assessed for clonality and were found to at least represent an oligoclonal population. Thus, mast cells and lymphocytes within focal aggregates in the bone marrow of those with mastocytosis are more frequently positive for the codon 816 activating mutation. Further, the B cell population is oligoclonal, suggesting that clonal proliferation is unlikely to be the basis of clustering.
authors.affiliation: Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 10 Center Drive, Room 11C 205, MSC1881, Bethesda, MD 20892-1881, USA.
authors.forename: Marcia L
authors.initials: ML
authors.lastname: Taylor
authors.forename: Devinder
authors.initials: D
authors.lastname: Sehgal
authors.forename: Mark
authors.initials: M
authors.lastname: Raffeld
authors.forename: Harold
authors.initials: H
authors.lastname: Obiakor
authors.forename: Cem
authors.initials: C
authors.lastname: Akin
authors.forename: Rose G
authors.initials: RG
authors.lastname: Mage
authors.forename: Dean D
authors.initials: DD
authors.lastname: Metcalfe
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D001065:Appendix / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology*; D001483:Base Sequence; D001706:Biopsy; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D049109:Cell Proliferation; D003001:Cloning, Molecular; D003062:Codon; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007834:Lasers; D008297:Male; D008407:Mast Cells / Q000166:cytology*; D008415:Mastocytosis / Q000235:genetics* / Q000473:pathology*; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics* / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000166:cytology*
chemical_list: D003062:Codon; D019009:Proto-Oncogene Proteins c-kit