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Transcripts in Subclones of an IgM+ Human B Cell Lymphoma Line1
Division of Life Sciences, Rutgers, The State University of New Jersey, Piscataway, NJ 08855
The CD40:CD40 ligand (CD40L) interaction plays a critical role in T
cell-dependent isotype switching. To elucidate the role of CD40
signaling in the activation of 
germline transcription and as an
extension, in targeting C regions for isotype switching, an
IgM+ Burkitt lymphoma cell line (Ramos 2G6) was
assayed for the up-regulation of germline transcripts after CD40L
stimulation. Independent Ramos 2G6 subclones that either expressed
(I+) or did not express (I-) basal
levels of I transcripts were assessed for their transcriptional
response to CD40L signaling by contact with either a Jurkat T cell line
(D1.1) or a transfected CD40L-expressing epithelial cell line
(293/CD40L) in the presence or absence of IL-4. Both I-
and I+ Ramos 2G6 subclones cultured with IL-4 and CD40L
markedly up-regulated germline transcription predominantly from the
1, 2, and 3 subclasses over levels obtained with IL-4 alone.
In addition, these two signals were required to obtain de novo switch
recombination. However, incubation with CD40L alone resulted in a
substantial increase in germline transcription only in the
I+ and not the I- subclones. Observed
basal transcription at the 1 locus also correlated with the ability
of not only the 1 locus, but also the 2 and 3 loci, to
up-regulate germline transcripts in response to CD40 signaling. These
data are consistent with CD40:CD40L contact up-regulating germline
transcription only after the B cell has received a signal that alters
the transcriptional state of the heavy chain locus.
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