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International Immunology, Vol. 11, No. 5, 753-763, May 1999
© 1999 Japanese Society for Immunology

An extrachromosomal switch recombination substrate reveals kinetics and substrate requirements of switch recombination in primary murine B cells

Katja Petry1, Gregor Siebenkotten1,3, Rainer Christine1,2, Katharina Hein1,2 and Andreas Radbruch1,2,4

1 Institut für Genetik der Universität zu Köln, 50931 Köln, Germany
2 Deutsches Rheumaforschungszentrum, Hannoversche Strasse 27, 10115 Berlin, Germany
3 AMAXA GmbH, 10117 Berlin, Germany
4 Medizinische Fakultät (Charité), Humboldt-Universität, 10098 Berlin, Germany

Correspondence to: A. Radbruch, Deutsches Rheumaforschungszentrum, Hannoversche Strasse 87, 10115 Berlin, Germany

Ig class switch recombination occurs in B lymphocytes upon activation, and is targeted to distinct switch (S) regions by cytokine-mediated induction of switch transcripts spanning the entire S region and the adjacent constant region gene segments. Using a novel type of switch recombination substrate, constructed according to the intron–exon structure of the IgH locus, but with heterologous elements, we here have tested the structural requirements for targeting and the kinetics of switch recombination in activated primary murine B cells. When transfected at various times after activation, up to 10% of the transfected B cells perform recombination of the substrate within 12 h. Switch recombination in primary B cells is restricted to the first 72 h after onset of activation, then rapidly decreases to background levels, as obtained in plasmacytoma cells or with substrates carrying no S region sequences. In terms of structural requirements, switch recombination is targeted to any transcription unit that contains an intronic S region and depends on processing of the primary transcript by splicing.

Keywords: B lymphocytes, class switch recombination, Ig, lipopolysaccharide, primary cells, transient transfection

Transmitting editor: M. Neuberger


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