title: Establishment and characterization of mouse bone marrow-derived mast cell hybridomas.
abstract: Interleukin (IL)-3-dependent mouse bone marrow-derived mast cells (BMMCs) are an important model for studying the function of mucosal-type mast cells. In the present study, BMMCs were successfully immortalized by cell fusion using a hypoxanthine-aminopterin-thymidine medium-sensitive variant of P815 mouse mastocytoma (P815-6TgR) as a partner cell line. The established mouse mast cell hybridomas (MMCHs) expressed α, β, and γ subunits of high-affinity immunoglobulin E (IgE) receptor (FcεRI) and possessed cytoplasmic granules devoid of or partially filled with electron-dense material. Four independent MMCH clones continuously proliferated without supplemental exogenous IL-3 and showed a degranulation response on stimulation with IgE+antigen. Furthermore, histamine synthesis and release by degranulation were confirmed in MMCH-D5, a MMCH clone that showed the strongest degranulation response. MMCH-D5 exhibited elevated levels of IL-3, IL-4, IL-13, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor (TNF)-α, and cyclooxygenase 2, and production of prostaglandin D(2) and leukotriene C(4) in response to IgE-induced stimulation. MMCH clones also expressed Toll-like receptors (TLRs) 1, 2, 4, and 6 and showed elevated levels of TNF-α expression in response to stimulation with TLR2 and TLR4 ligands. The MMCHs established using this method should be suitable for studies on FcεRI- and TLR-mediated effector functions of mast cells.
authors: Integrated Department of Sciences of Functional Foods, Graduate School of Agriculture, Shinshu University, Nagano, Japan. tkawafb@shinshu-u.ac.jp
authors: Takeshi
authors: T
authors: Kawahara
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D015550:Cell Degranulation; D006825:Hybridomas / Q000166:cytology* / Q000378:metabolism; D008024:Ligands; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D011961:Receptors, Fc / Q000235:genetics / Q000378:metabolism; D017455:Receptors, IgE / Q000276:immunology / Q000378:metabolism; D051193:Toll-Like Receptors / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C036832:Fc(alpha) receptor; D008024:Ligands; D011961:Receptors, Fc; D017455:Receptors, IgE; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha