I would like to activate populations of B cells via CD40 ligation (i.e.
using anti-CD40 mAb), but am confused somewhat by information in the
available literature... Is adding anti-CD40 mAb to the culture system
sufficient, or must the molecule be cell bound?? My references all seem to
say that a transfected cell line was created to carry the CD40L?? Why is
this necessary? I have concluded that IL-4 or an additional stimulus (SAC)
are necessary as a "second messenger" in this system. Would calcium
ionophore do the same thing?
And why when using anti-Ig to stimulate does it have to be bound to
something (such as polyacrylamide beads)?
Why would I NOT want to use whole blood when activating B cells? I realize
sorted or magnetic-bead purified B cells would avoid stimulation from other
cell populations, but if I am interested in coming as close as possible to
an "in vivo" model??.....
Any experience that ANYONE has had with activating B cells in vitro would
be appreciated, and references would also be helpful.
I do appreciate all the "hand-holding" that the scientists here have
provided in the past, and once again apologize for my bumbling questions.
Thanks again,
-- Keith Bahjat Northwestern University Medical School Comprehensive AIDS Center Chicago, IL Kbahjat@nwu.edu