Evidence for Class-specific Factors in Immunoglobulin Isotype Switching

doi:10.1084/jem.191.8.1365

Published online 18 April 2000.

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© The Rockefeller University Press, 0022-1007/2000/4/1365/ $5.00
The Journal of Experimental Medicine, Volume 191, Number 8, April 17, 2000 1365-1380

Original Article

Evidence for Class-specific Factors in Immunoglobulin Isotype Switching

Ananth Shanmugam^a, Meng-Jiao Shi^b, Lauren Yauch^a, Janet Stavnezer^b, and Amy L. Kenter^a
^a Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago, Illinois 60612
^b Department of Molecular Genetics and Microbiology and the Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, Massachusetts 01655

Correspondence to: Amy L. Kenter, Department of Microbiology/Immunology (M/C790), College of Medicine, University of Illinois, Chicago, IL 60612-7344. Tel:312-996-5293 Fax:312-996-6415 E-mail:amy.l.kenter{at}uic.edu.

Immunoglobulin class switch recombination (SR) occurs by a Bcell–specific, intrachromosomal deletional process betweenswitch regions. We have developed a plasmid-based transienttransfection assay for SR to test for the presence of transactingswitch activities. The plasmids are novel in that they lacka eukaryotic origin of DNA replication. The recombination activityof these switch substrates is restricted to a subset of B celllines that support isotype switching on their endogenous lociand to mitogen-activated normal splenic B cells. The factorsrequired for extrachromosomal plasmid recombination are constitutivelyexpressed in proliferating splenic B cells and in B cell linescapable of inducibly undergoing immunoglobulin SR on their chromosomalgenes. These studies suggest that mitogens that induce switchingon the chromosome induce accessibility rather than switch recombinaseactivity. Finally, we provide evidence for two distinct switchingactivities which independently mediate µ $->$ ${alpha}$ and µ $->$ ${gamma}$ 3SR.

Key Words: immunoglobulin, isotype switch, plasmid assay, transient transfection,PCR

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