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Induced B Cell Development in Adult Mice.
Brennecke, Anne-Margarete Düber, Sandra Roy, Bishnudeo Thomsen, Irene Garbe, Annette I Pabst, Oliver Kretschmer, Karsten Weiss, Siegfried
Brennecke, Anne-Margarete
Düber, Sandra
Roy, Bishnudeo
Thomsen, Irene
Garbe, Annette I
Pabst, Oliver
Kretschmer, Karsten
Weiss, Siegfried
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2018-01-01
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We employed the B-Indu-Rag1 model in which the coding exon of recombination-activating gene 1 (Rag1) is inactivated by inversion. It is flanked by inverted loxP sites. Accordingly, B cell development is stopped at the pro/pre B-I cell precursor stage. A B cell-specific Cre recombinase fused to a mutated estrogen receptor allows the induction of RAG1 function and B cell development by application of Tamoxifen. Since Rag1 function is recovered in a non-self-renewing precursor cell, only single waves of development can be induced. Using this system, we could determine that B cells minimally require 5 days to undergo development from pro/preB-I cells to the large and 6 days to the small preB-II cell stage. First immature transitional (T) 1 and T2 B cells could be detected in the bone marrow at day 6 and day 7, respectively, while their appearance in the spleen took one additional day. We also tested a contribution of adult bone marrow to the pool of B-1 cells. Sublethally irradiated syngeneic WT mice were adoptively transferred with bone marrow of B-Indu-Rag1 mice and B cell development was induced after 6 weeks. A significant portion of donor derived B-1 cells could be detected in such adult mice. Finally, early VH gene usage was tested after induction of B cell development. During the earliest time points the VH genes proximal to D/J were found to be predominantly rearranged. At later time points, the large family of the most distal VH prevailed.
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1664-3224
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Attribution-NonCommercial-ShareAlike 4.0 International