International Immunology, Vol 10, 1027-1037, Copyright © 1998 by Oxford University Press
K Cunningham, H Ackerly, L Claflin, J Collins, P Wu, C Ford, R Lansford, F Alt and WA Dunnick
To investigate the regulation of Ig switch recombination, we have
constructed mice with a 56 kb VDJmudeltagamma1 transgene. This transgene
included an anti-nitrophenyl VDJ segment, Smu, Cmu, Cdelta, Igamma1,
Sgamma1, Cgamma1 and the Cgamma1 membrane exons from the murine Igh(a)
haplotype. Two founder lines were produced, with very similar
characteristics. Transgenic B cells expressed normal amounts of Cmu (which
is >95% transgenic), Cdelta and other cell surface markers, and normal
amounts of VDJ and Cmu RNA. Gamma1 germline transcription of the transgenes
is properly regulated since stable transcripts were not expressed in B
cells treated with lipopolysaccharide (LPS) alone, nor in thymus or
non-lymphoid tissues, but were expressed after treatment of B cells with
LPS + IL-4 or CD40L + IL-4. B cells from both lines of transgenic mice
expressed transgenic gamma1a after in vitro culture with CD40L + IL-4, but
not after culture with CD40L alone. However, the CD40L + IL-4 induced IgG1
precursor frequency is much lower for VDJmudeltagamma1 transgenic B cells
(1:240-760) than for non-transgenic B cells (1:9). Analysis of DNA from
transgenic hybridomas indicated that switch recombination can take place in
switch (S) regions, but can also take place outside S regions. These
results indicate that targeting of switch recombinase to S regions must
include regulation beyond the S regions themselves and correct germline
transcription. This additional regulation might include cis-acting elements
or appropriate spacing or arrangement of the recombining elements.
ARTICLES
Germline transcription and recombination of a murine VDJmudeltagamma1 transgene
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109-0620, USA.
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