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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
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web