From: Keith Bahjat (Kbahjat@nwu.edu)
Date: Thu Dec 26 1996 - 16:36:27 EST
Dear Gods of All that Flows, 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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