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Journal of Biochemistry Advance Access published online on July 5, 2006
Journal of Biochemistry, doi:10.1093/jb/mvj140
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© 2006 The Japanese Biochemical Society.
Received March 23, 2006
Accepted May 31, 2006

Regular Paper

Novel Mast Cell Lines with Enhanced Proliferative and Degranulative Abilities Established from Temperature-Sensitive SV40 Large T Antigen Transgenic Mice

Masahiko Kanehira 1, Tomonori Kaifu 1, Kozue Maya 1, Mitsuji Kaji 2, Akira Nakamura 1, Masuo Obinata 3, and Toshiyuki Takai 1 *

1 Department of Experimental Immunology and the Core Research for Evolutional Science and Technology (CREST) Program of the Japan Science and Technology Agency (JST), Tohoku University, Sendai 980-8575, Japan
2 Common Instrument Center, Tohoku University, Sendai 980-8575, Japan
3 Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University, Sendai 980-8575, Japan

* To whom correspondence should be addressed.
Toshiyuki Takai, E-mail: tostakai{at}idac.tohoku.ac.jp


  Abstract

Mast cells (MCs) play crucial roles in innate immunity to parasitic and bacterial infections as well as in hypersensitivity, such as the induction and exacerbation of allergy and autoimmune diseases. The regulatory mechanisms for MC development and effector functions are of great interest for developing novel therapeutic strategies against such disorders. Here we report the establishment of novel, immortalized MC lines from bone marrow (BM) cells of a temperature-sensitive mutant of SV40 large T antigen-transgenic mice (termed SVMCs). BM cells from tsSV40LT mice were cultured in the presence of interleukin (IL)-3 for 3 weeks, and then subjected to limiting dilution and single-cell cloning, yielding 27 independent MC clones, three of which were subjected to further analysis. On culture with nerve growth factor, stem cell factor and IL-3, these SVMC clones showed morphologic and biochemical changes from mucosal MC-like to connective-tissue MC-like phenotypes. These SVMC lines exhibited a significantly enhanced proliferation rate, and a higher responsiveness to the high affinity Fc receptor for IgE-mediated intracellular calcium mobilization and degranulation than those of BM-derived cultured MCs. These cell lines should facilitate studies on the mechanisms for the development, differentiation and effector functions of MCs in health and diseases.


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