The Journal of Experimental Medicine
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Published online 6 August 2001. doi:10.1084/jem.194.3.247
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© The Rockefeller University Press, 0022-1007/2001/8/247/ $5.00
The Journal of Experimental Medicine, Volume 194, Number 3, August 6, 2001 247-254

Original Article

Transplanted Long-Term Cultured Pre-Bi Cells Expressing Calpastatin Are Resistant to B Cell Receptor–Induced Apoptosis

Antonio Ruiz-Velaa, Fernando Serranoa, Manuel A. Gonzáleza, José Luis Abada, Antonio Bernada, Masatoshi Makib, and Carlos Martínez-Aa

a Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Universidad Autónoma de Madrid, Campus de Cantoblanco, E-28049 Madrid, Spain
b Department of Applied Molecular Biosciences, Nagoya University, Nagoya 464-8601, Japan
Dept. Immunology and Oncology, Centro Nacional de Biotecnología CSIC, Campus de Cantoblanco UAM, E-28049 Madrid, Spain.34-91-372-049334-91-585-4537

aruiz{at}cnb.uam.es

Long-term cultured pre-B cells are able to differentiate into immunoglobulin (Ig)M-positive B cells (IgM+ cells) when transplanted into severe combined immunodeficient (SCID) mice. Based on previous studies, here we report the development of a reconstitution assay in nonobese diabetic/SCID (NOD/SCID) mice using pre-B cells, which allows us to study the role of calpains (calcium-activated endopeptidases) during B cell development as well as in B cell clonal deletion. Using this model, we show that calpastatin (the natural inhibitor of calpains) inhibits B cell receptor–induced apoptosis in IgM+ cells derived from transplanted mice. We thus hypothesize an important function for calpain in sculpting the B cell repertoire.

Key Words: pre-B cells • calpastatin • calpain • BCR • transplant

Abbreviations used in this paper: Act-D, actinomycin D; BCR, B cell receptor; BM, bone marrow; GFP, green fluorescence protein; IRES, internal ribosome entry site; NOD, nonobese diabetic; SPRD, streptavidin-SpectralRed.

© 2001 The Rockefeller University Press


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